Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermi...Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermin D(GSDMD)cleavage plays a pivotal role in disease progression through nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP-3)/caspase-1 classical and caspase-4/5/11 non-classical pathways.Targeting pyroptosis has emerged as a promising therapeutic strategy,with recent advances highlighting the potential of pyroptosis inhibitors such as small-molecule compounds,biologics,and repurposed drugs that specifically target NLRP3,caspases,or GSDMD to suppress inflammasome activation,block pore formation,and mitigate downstream inflammation.This review systematically summarizes the mechanisms and therapeutic effects of these inhibitors,while addressing critical challenges including drug specificity,delivery efficiency,and long-term safety,and explores their potential in combination therapies with existing IBD treatments to enhance clinical efficacy.By integrating preclinical and clinical evidence,we provide valuable insights into the translational prospects of pyroptosis-targeted therapies for precision management of IBD.展开更多
Regulated cell death is a form of cell death that is actively controlled by biomolecules.Several studies have shown that regulated cell death plays a key role after spinal cord injury.Pyroptosis and ferroptosis are ne...Regulated cell death is a form of cell death that is actively controlled by biomolecules.Several studies have shown that regulated cell death plays a key role after spinal cord injury.Pyroptosis and ferroptosis are newly discovered types of regulated cell deaths that have been shown to exacerbate inflammation and lead to cell death in damaged spinal cords.Autophagy,a complex form of cell death that is interconnected with various regulated cell death mechanisms,has garnered significant attention in the study of spinal cord injury.This injury triggers not only cell death but also cellular survival responses.Multiple signaling pathways play pivotal roles in influencing the processes of both deterioration and repair in spinal cord injury by regulating pyroptosis,ferroptosis,and autophagy.Therefore,this review aims to comprehensively examine the mechanisms underlying regulated cell deaths,the signaling pathways that modulate these mechanisms,and the potential therapeutic targets for spinal cord injury.Our analysis suggests that targeting the common regulatory signaling pathways of different regulated cell deaths could be a promising strategy to promote cell survival and enhance the repair of spinal cord injury.Moreover,a holistic approach that incorporates multiple regulated cell deaths and their regulatory pathways presents a promising multi-target therapeutic strategy for the management of spinal cord injury.展开更多
Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to impr...Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.展开更多
Bisphenol A(BPA)is an industrial pollutant that can cause immune impairment.Selenium acts as an antioxidant,as selenium deficiency often accompanies oxidative stress,resulting in organ damage.This study is the first t...Bisphenol A(BPA)is an industrial pollutant that can cause immune impairment.Selenium acts as an antioxidant,as selenium deficiency often accompanies oxidative stress,resulting in organ damage.This study is the first to demonstrate that BPA and/or selenium deficiency induce pyroptosis and ferroptosis-mediated thymic injury in chicken and chicken lymphoma cell(MDCC-MSB-1)via oxidative stress-induced endoplasmic reticulum(ER)stress.We established a broiler chicken model of BPA and/or selenium deficiency exposure and collected thymus samples as research subjects after 42 days.The results demonstrated that BPA or selenium deficiency led to a decrease in antioxidant enzyme activities(T-AOC,CAT,and GSH-Px),accumulation of peroxides(H2O2 and MDA),significant upregulation of ER stress-relatedmarkers(GRP78,IER 1,PERK,EIF-2α,ATF4,and CHOP),a significant increase in iron ion levels,significant upregulation of pyroptosis-related gene(NLRP3,ASC,Caspase1,GSDMD,IL-18 and IL-1β),significantly increase ferroptosis-related genes(TFRC,COX2)and downregulate GPX4,HO-1,FTH,NADPH.In vitro experiments conducted in MDCC-MSB-1 cells confirmed the results,demonstrating that the addition of antioxidant(NAC),ER stress inhibitor(TUDCA)and pyroptosis inhibitor(Vx765)alleviated oxidative stress,endoplasmic reticulum stress,pyroptosis,and ferroptosis.Overall,this study concludes that the combined effects of oxidative stress and ER stress mediate pyroptosis and ferroptosis in chicken thymus induced by BPA exposure and selenium deficiency.展开更多
Background Oxidative stress significantly impacts growth performance and liver function in piglets.Ferulic acid(FA)works as an antioxidant,however,the role and mechanism of FA in the regulation of diquat-induced oxida...Background Oxidative stress significantly impacts growth performance and liver function in piglets.Ferulic acid(FA)works as an antioxidant,however,the role and mechanism of FA in the regulation of diquat-induced oxidative stress in piglets are less known.This study was designed to investigate the effects of FA on growth performance and antioxi-dant capacity in piglets with diquat challenge.Methods Thirty-two healthy DLY(Duroc×Landrace×Yorkshire)piglets(13.24±0.19 kg)were randomly divided into one of two diets including 0 or 4 g/kg FA for 14 d.On d 15,all pigs were intraperitoneally injected diquat or sterile saline.Results Dietary supplementation with ferulic acid(FA)significantly improved the average daily gain(ADG)and decreased feed-gain ratio(F/G)of piglets.Here,dietary FA supplementation reduced serum aspartate aminotrans-ferase(AST),alanine aminotransferase(ALT)activities in diquat challenged piglets.Furthermore,diquat infusion increased reactive oxygen radicals(ROS)level in liver,decreased the activities of total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC)and increased malondialdehyde(MDA)con-tent in the liver and serum.Supplementation with FA significantly increased T-AOC and T-SOD activities and decreased MDA and ROS levels.FA down-regulated gene and protein expression of Keap1,and up-regulated protein expression of Nrf2 and HO-1 in the liver of piglets with diquat challenge.Importantly,diquat challenge increased the ratio of late apoptosis,increased serum levels of IL-1β,IL-18 and lactate dehydrogenase(LDH),and up-regulated pyroptosis-related genes in the liver.FA supplementation reduced the ratio of late apoptosis and down-regulated mRNA expression of Caspase-1.Accordingly,FA addition reduced concentration of IL-1β,IL-18,and LDH under diquat challenge.Conclusions Diquat-induced oxidative stress reduced growth performance and impaired liver function in piglets.Dietary FA supplementation enhanced the antioxidant capacity and reduced the degree of hepatocyte pyroptosis,thereby alleviating the oxidative damage in the liver and mitigating the impact of diquat on growth performance of piglets.展开更多
Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that ...Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.展开更多
The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economic...The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.展开更多
Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisp...Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.展开更多
Background:The Shenlian formula demonstrates therapeutic bmodelenefits for diabetic kidney disease(DKD).Yet,its effectiveness in mitigating renal interstitial fibrosis(RIF)and the pharmacological underpinnings remain ...Background:The Shenlian formula demonstrates therapeutic bmodelenefits for diabetic kidney disease(DKD).Yet,its effectiveness in mitigating renal interstitial fibrosis(RIF)and the pharmacological underpinnings remain to be elucidated.This investigation seeks to delineate some of the formula’s potential mechanisms via experimental validation.Methods:The study initiated by inducing a DKD in rats through unilateral nephrectomy combined with streptozotocin(STZ)administration and an advanced glycation end products-bovine serum albumin(AGE-BSA)induced DKD model in HK-2 cells to assess the Shenlian formula’s renal protective effects.Renal tissues underwent pathological and immunohistochemical staining to evaluate improvements in RIF.Mechanistic insights were further obtained through techniques such as Western Blot,immunofluorescence,co-immunoprecipitation,enzyme-linked immunosorbent assay(ELISA),and protein docking analyses.Results:Shenlian formula could enhance renal function,alleviate tubular damage,suppress epithelial-mesenchymal transition(EMT),reduce extracellular matrix(ECM)deposition,and thus decelerate RIF progression.It notably decreased the expression of markers associated with pyroptosis(NOD-,LRR-and pyrin domain-containing protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),caspase1,cleaved-caspase1,gasdermin D(GSDMD),GSDMD-N,interleukin-18(IL-18),interleukin-1 beta(IL-1β)),effectively inhibiting renal cell pyroptosis.Moreover,the formula facilitated autophagy substrate(sequestosome 1(p62))degradation,efficiently restoring the autophagy pathway.It also modulated autophagy,impacting cell pyroptosis regulation.Conclusion:The Shenlian formula potentially inhibits cellular pyroptosis by modulating the autophagy pathway,thereby diminishing inflammation-induced renal injury and fibrosis.This finding suggests a novel therapeutic approach for managing DKD-induced fibrosis.展开更多
Background:Activation of NLRP3(NOD-,LRR-and pyrin domain-containing protein 3)inflammasomes induced by pyroptosis is crucial in metabolic dysfunction-associated steatohepatitis(MASH)progression.Adiponectin possesses a...Background:Activation of NLRP3(NOD-,LRR-and pyrin domain-containing protein 3)inflammasomes induced by pyroptosis is crucial in metabolic dysfunction-associated steatohepatitis(MASH)progression.Adiponectin possesses an anti-inflammatory role in various liver diseases.This study aimed to evaluate the effects of adiponectin on MASH.Methods:Adiponectin-mediated anti-inflammatory mechanisms,effects on pyroptosis-related proteins,and activation of NLRP3 inflammasomes were investigated using methionine-choline-deficient(MCD)-induced MASH murine model and in vitro models.The degree of MASH inflammation in liver tissue of C57BL/6J mice was assessed using histopathology.Enzyme-linked immunosorbent assay was performed to measure levels of inflammatory factors[interleukin-18(IL-18),IL-1β,and tumor necrosis factor-α(TNF-α)]in mice serum and culture medium.Western blot and quantitative polymerase chain reaction were performed to analyze the expression of pyroptosis-related genes and proteins in liver tissues of mouse model and in vitro models.Macrophage recruitment in vitro was evaluated using co-culture of upper and lower chambers.Results:MASH developed in MCD diet mice[metabolic dysfunction-associated steatotic liver disease(MASLD)activity score=6]but not in methionine-choline-sufficient(MCS)diet mice(MASLD activity score=3).Compared to MCS-fed mice,MCD-fed mice showed increased serum levels of aspartate amino-transferase,IL-18,IL-1β,and TNF-αand higher MASLD activity score(P<0.001).Adiponectin inhibited these increases(P<0.05)and suppressed mRNA and protein levels of NLRP3,gasdermin-D(GSDMD),and GSDMD-N in liver tissues(P<0.05).In vitro,lipopolysaccharide(LPS)/palmitic acid(PA)increased the levels of IL-18,IL-1β,and TNF-α,mRNA expressions of CASP1 and GSDMD,and production of CASP1,NLRP3,GSDMD,and GSDMD-N(P<0.01).Adiponectin reduced the levels of these inflammatory fac-tors and downregulated the mRNA expression and protein generation of pyroptosis-related markers(P<0.05).HepG2 cells pretreated with LPS/PA recruited more J774A.1 cells(P<0.001)and increased inflam-matory factor secretion by J774A.1 cells(P<0.001).Adiponectin inhibited this recruitment and reduced inflammatory factor secretion(P<0.001).Conclusions:Adiponectin inhibits hepatocyte pyroptosis by reducing the production and activation of NLRP3 inflammasomes,CASP1,and GSDMD,thus improving the inflammatory response in MASH and possibly delaying or reversing MASLD progression.展开更多
To the Editor:Pancreatic cancer is a malignancy characterized by a poor prog-nosis,with a 5-year survival rate of<10%[1].Furthermore,only a minority of patients(<20%)qualify for curative-intent resec-tion,and ev...To the Editor:Pancreatic cancer is a malignancy characterized by a poor prog-nosis,with a 5-year survival rate of<10%[1].Furthermore,only a minority of patients(<20%)qualify for curative-intent resec-tion,and even among those who undergo this procedure,the risk of recurrence within three years remains alarmingly high,reach-ing up to 70%[2].Due to the lack of specific clinical manifes-tations of pancreatic cancer,most cases have metastasized or in-vaded the major vessels around the pancreas at the time of initial diagnosis,resulting in a low surgical resection rate.Even patients who undergo surgical resection often face a poor prognosis[3].In recent years,neoadjuvant chemotherapy using agents such as gemcitabine,5-fluorouracil,albumin-bound paclitaxel,modified fluorouracil/leucovorin plus irinotecan,and oxaliplatin(mFOLFIRI-NOX),targeted therapies addressing molecular subtypes of pan-creatic cancer,and immunotherapies targeting PD-1 and PD-L1 have shown efficacy in improving the overall prognosis of patients with pancreatic cancer,although the impact remains modest[4,5].Therefore,novel therapeutic strategies and prognostic evaluation systems are urgently needed to enhance the survival of patients with pancreatic cancer.展开更多
BACKGROUND Pyroptosis and ubiquitination have been identified as key processes influencing the pathogenesis of diabetes mellitus(DM).AIM To investigate the genes associated with the ubiquitin-proteasome system(UPS)and...BACKGROUND Pyroptosis and ubiquitination have been identified as key processes influencing the pathogenesis of diabetes mellitus(DM).AIM To investigate the genes associated with the ubiquitin-proteasome system(UPS)and pyroptosis in type 2 DM(T2DM),and elucidate their mechanisms of action in T2DM.METHODS The datasets GSE76894,GSE41762,and GSE86469 were utilized in this study.UPS-related genes(UPSGs)and pyroptosis-related genes(PRGs)were obtained from existing literature.Differential expression analysis was performed to identify differentially expressed genes(DEGs).DEGs were intersected with UPSGs and PRGs to identify differentially expressed UPSGs and PRGs.Ubiquitin-pyroptosisrelated biomarkers were determined using Spearman’s correlation,t-tests,and receiver operating characteristic curve analysis.Pathway enrichment of biomarkers was assessed using Gene Set Enrichment Analysis(GSEA).Single sample GSEA(ssGSEA)and Spearman’s correlation were used to analyze the relationship between biomarkers and immune cells.A competitive endogenous RNA network was constructed.Subsequently,drugs related to the biomarkers were identified and a gene-drug network was established.In dataset GSE86469,single-cell sequencing was utilized to determine cell types.Finally,the expression levels of biomarkers were validated through quantitative PCR(qPCR)and western blot analysis.RESULTS A total of 581 DEGs were identified in GSE76894.Four genes[ATP binding cassette subfamily C member 8(ABCC8),retinol binding protein 4(RBP4),Ras protein-specific guanine nucleotide-releasing factor 1(RASGRF1),and solute carrier family 34 member 2(SLC34A2)]were identified as ubiquitin-pyroptosis-related biomarkers in T2DM,based on consistent expression trends and significant differences in GSE76894 and GSE41762.These biomarkers were enriched in oxidative phosphorylation and mitogen-activated protein kinase signaling pathways,which are relevant to DM.ssGSEA revealed significant differences in the enrichment scores of nine immune cell types between groups.A total of 17 microRNAs(miRNAs)and 36 long non-coding RNAs(lncRNAs)were identified,forming numerous miRNA-lncRNA interactions.Additionally,22 drugs related to the biomarkers,such as gliclazide and tretinoin,were identified.In GSE86469,eight cell types,including alpha and beta cells,were characterized.qPCR and western blot analysis confirmed that the expression trends of RASGRF1 and SLC34A2 were consistent with the findings in GSE76894.CONCLUSION This study identified four ubiquitin-pyroptosis-related biomarkers(ABCC8,RBP4,RASGRF1,and SLC34A2)in T2DM through bioinformatics analysis,providing novel insights into the diagnosis and treatment of T2DM.展开更多
Sustained inflammatory responses are closely related to various severe diseases,and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment.Natural produ...Sustained inflammatory responses are closely related to various severe diseases,and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment.Natural products have garnered considerable concern for the treatment of inflammation.Huanglian-Wumei decoction(HLWMD)is a classic prescription used for treating inflammatory diseases,but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated.Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory,we successfully obtained berberine(BBR)-chlorogenic acid(CGA)supramolecular(BCS),which is an herbal pair from HLWMD.Using a series of characterization methods,we confirmed the self-assembly mechanism of BCS.BBR and CGA were self-assembled and stacked into amphiphilic spherical supramolecules in a 2:1 molar ratio,driven by electrostatic interactions,hydrophobic interactions,andπ–πstacking;the hydrophilic fragments of CGA were outside,and the hydrophobic fragments of BBR were inside.This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules.Compared with free molecules,BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide(LPS)-induced pyroptosis.Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB(NF-κB)p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.展开更多
Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative diseases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and t...Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative diseases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing.There are currently no effective strategies for slowing the progression of AD.Herein,we spotlight the dysregulation of lipid metabolism,particularly the elevation of ceramides(Cers),as a critical yet underexplored facet of AD pathogenesis.Our study delineates the role of Cers in promoting microglial pyroptosis,a form of programmed cell death distinct from apoptosis and necroptosis,characterized by cellular swelling,and membrane rupture mediated by the NLRP3 inflammasome pathway.Utilizing both in vivo experiments with amyloid precursor protein(APP)/presenilin 1(PS1)transgenic mice and in vitro assays with BV-2 microglial cells,we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin(ICA),a flavonoid with known antioxidant and anti-inflammatory properties.Our findings reveal a significant increase in Cers levels and pyroptosis markers(NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain,caspase-1,gasdermin D(GSDMD),and interleukin-18(IL-18))in the brains of AD model mice,indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis.Conversely,ICA treatment effectively reduces these pyroptotic markers and Cer levels,thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD.This study not only advances our understanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy,capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2(COX-2)-NLRP3 inflammasome-gasdermin D(GSDMD)axis.Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of targeting microglial pyroptosis in AD.展开更多
Background:Atherosclerosis(AS),the primary pathological foundation of cardiovascular diseases,is characterized by intricate processes including inflammation,lipid metabolism disorders,and pyroptosis.While the traditio...Background:Atherosclerosis(AS),the primary pathological foundation of cardiovascular diseases,is characterized by intricate processes including inflammation,lipid metabolism disorders,and pyroptosis.While the traditional Chinese medicine compound Dingxin Recipe(DXR)has demonstrated definitive clinical efficacy in treating AS,its therapeutic mechanisms remain unclear.This study employed an integrated approach combining network pharmacology,molecular docking,and molecular dynamics simulations(MDS)to investigate DXR’s anti-AS mechanisms.Methods:Active ingredients and targets of DXR were identified and screened using databases such as GeneCards,OMIM,and TCMSP.An“ingredient-target-disease”network was constructed to visualize these interactions.Molecular docking was utilized to assess the binding affinity between key ingredients and their respective targets.Additionally,MDS were conducted to analyze the stability of these complexes,providing robust evidence for further clinical applications and in-depth research.Results:Through network pharmacology analysis,we identified 99 active drug components,934 gene targets,and 1463 disease targets associated with DXR.Protein-protein interaction analysis revealed central regulatory nodes.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that these components primarily modulate processes such as inflammatory response and transcription factor activation,and are closely linked to the AGERAGE signaling pathway,lipid metabolism,and atherosclerosis pathways.Molecular docking confirmed strong binding potential between the components and their targets,while MDS further validated the stability of these interactions.Conclusion:This study elucidates that the active ingredients in DXR alleviate AS by mitigating inflammatory responses and inhibiting pyroptosis through the suppression of inflammatory factor release.These findings provide a scientific foundation for the clinical application of DXR in AS treatment.展开更多
Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiolog...Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiological factors have been shown to cause acquired SNHL,such as ototoxic drugs,noise exposure,aging,infections,and diseases.Several programmed cell death(PCD)pathways have been reported to be involved in SNHL,especially some novel PCD pathways that have only recently been reported,such as ferroptosis,necroptosis,and pyroptosis.Here we summarize these PCD pathways and their roles and mechanisms in SNHL,aiming to provide new insights and potential therapeutic strategies for SNHL by targeting these PCD pathways.展开更多
Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI....Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI.However,the mechanism remains unclear.This study aimed to investigate whether c-Myc affects CIRI by regulating Serine/Arginine-rich Splicing Factor 1(SRSF1)/NLRP1-mediated pyroptosis.Methods:Oxygen-glucose deprivation/reperfusion(OGD/R)induced neuroblastoma cells for the establishment of an in vitro CIRI model.The levels of c-Myc and SRSF1,cell viability,the expression of pyroptosis-related factors,and the interaction between SRSF1 and NLRP1 were evaluated.Results:The expression of c-Myc and SRSF1 was decreased in OGD/R-induced neuroblastoma cells.c-Myc overexpression increased c-Myc and SRSF1 expression and cell viability in OGD/Rinduced neuroblastoma cells while inhibiting NLRP1,Caspase1,apoptosis-associated speck-like protein containing a CARD(ASC),interleukin-1beta(IL-1β),IL-18,and lactate dehydrogenase levels and pyroptosis.C-Myc was positively correlated with SRSF1.SRSF1 low expression reversed the effects of c-Myc on the above indicators in OGD/Rinduced neuroblastoma cells.Mechanically,SRSF1 interacted with NLRP1.SRSF1 was negatively correlated with NLRP1.The NLRP1 activator muramyl dipeptide(MDP)reversed the SRSF1 effect on OGD/R-induced neuroblastoma cells.Conclusion:Our results indicated that c-Myc reduced OGD/R-induced neuroblastoma cell pyroptosis by inhibiting NLRP1 activation by positive feedback SRSF1 signal.Our findings suggested that the c-Myc/SRSF1 axis might be a new strategy for treating CIRI in the clinic.展开更多
Background:Acute respiratory distress syndrome(ARDS)is the major therapeutic dilemma associated with significant inflammation and severe pulmonary dysfunction.Liriodendrin is a bioactive compound extract from traditio...Background:Acute respiratory distress syndrome(ARDS)is the major therapeutic dilemma associated with significant inflammation and severe pulmonary dysfunction.Liriodendrin is a bioactive compound extract from traditional Chinesemedicine,historically utilized formodulating inflammatory responses and alleviating symptoms in multiple diseasemodels.Methods:At present,BALB/c mice to explore the effects of liriodendrin on lipopolysaccharide(LPS)-induced ARDS.Before LPS was administered,the mice were treated with either liriodendrin or dexamethasone.Leukocyte infiltration,lung edema,and alveolar-capillary barrier integrity were evaluated in the bronchoalveolar lavage fluid(BALF)and pulmonary parenchyma.The expression of adhesion molecules and proinflammatory cytokines in BALF was evaluated by enzyme-linked immunosorbent assay.Western blotting assay facilitated the analysis of the expression or phosphorylation of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),NOD-like receptor family pyrin domain-containing 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),cleaved caspase-1(CL-csapase-1),nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB),inhibitor of kappa B(IκB),mitogen-activated protein kinase(MAPK),and protein kinase B(Akt)in the lungs.In addition,the anti-inflammatory effects of liriodendrin were evaluated in LPS-stimulated RAW264.7 macrophages.Before LPS was administered,the RAW264.7 macrophages were treated with either liriodendrin or dexamethasone.Nitric Oxide(NO)production was measured using the Griess reaction assay,while ELISA assessed IL-1β,IL-6,and TNF-αlevels.Western blot analysis evaluated NF-κB phosphorylation and the expression of NLRP3,ASC,and CLcaspase-1.Results:These outcomes revealed that liriodendrin intervention markedly ameliorated the pathological features of LPS-induced ARDS,including leukocyte infiltration,lung edema,and alveolar-capillary barrier disruption.Liriodendrin also reduced the LPS-induced secretion of intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesionmolecule-1(VCAM-1),expression of iNOS and COX2,and production of proinflammatory cytokines.Finally,we further discovered that the concentration trend of liriodendron amelioration ofARDSwas similar to those ofNLRP3 formation,NF-κB pathway activation,and p38 MAPK,c-Jun N-terminal kinase(JNK),and Akt phosphorylation but not to that of extracellular signal-regulated kinase(ERK)phosphorylation.Liriodendrin inhibited LPS-induced inflammatory responses in RAW264.7 macrophages.It markedly reduced NO production,propro-inflammatorytokines,NF-κB phosphorylation,and NLRP3 formation.Conclusions:In summary,liriodendrin effectively ameliorated the pathological features of LPS-induced ARDS inmice,demonstrating significant anti-inflammatory properties attributed to NLRP3 formation through NF-κB pathway activation by p38MAPK,JNK,and Akt phosphorylation.In LPS-treated RAW264.7 macrophages,liriodendrin reduced NO production,pro-inflammatory cytokines,and NLRP3 formation,suggesting its potential as an agent for ARDS and relative inflammation.展开更多
Background Deoxynivalenol(DON)is a mycotoxin that severely pollutes feed ingredients,and methods for reducing DON toxicity have become a significant research direction.Chlorogenic acid(CGA)is an active polyphenol foun...Background Deoxynivalenol(DON)is a mycotoxin that severely pollutes feed ingredients,and methods for reducing DON toxicity have become a significant research direction.Chlorogenic acid(CGA)is an active polyphenol found in some plants,which has anti-inflammatory and antioxidant properties and a protective effect on animal intestinal health.The effects of CGA on DON-induced pyroptosis in the intestinal porcine epithelial cell line-J2(IPEC-J2)and its potential mechanism were explored in this study.Results IPEC-J2 cells viability and membrane integrity were inversely correlated with DON concentration.Compared to those in the group treated with DON alone at 2,500 ng/mL,pretreatment with 80μmol/L CGA for 4 h significantly improved cell viability(P<0.01),and the alleviation of typical pyroptotic symptoms induced by DON were observed,including reduced cellular DNA fragmentation,decreased release of lactate dehydrogenase(LDH),normalized ROS levels,restoration of extracellularCa2+andK+contents to normal levels(P<0.01),as well as suppressed the enzyme activities of caspase-1 and caspase-4(P<0.01).Additionally,the mRNA expression levels of TNF,MDP,NOD2,TLR4,ASC and GSDMD were significantly improved(P<0.01),while both mRNA and protein expression levels of NF-κB,NLRP3,caspase-1,IL-1βand IL-18 were significantly upregulated(P<0.01)in the CGA+DON group,compare to those in the DON group.Conclusion Pretreatment with 80μmol/L CGA for 4 h effectively alleviated pyroptosis in IPEC-J2 cells induced by 2,500 ng/mL of DON through inhibiting activation of the NF-κB/NLRP3/capase-1 pathway.展开更多
AIM:To investigate whether pyroptosis contributes to retinal ganglion cell(RGC)degeneration in aged TgAPPswePS1 transgenic mice and to explore the relationship between amyloid-beta(Aβ)accumulation and activation of t...AIM:To investigate whether pyroptosis contributes to retinal ganglion cell(RGC)degeneration in aged TgAPPswePS1 transgenic mice and to explore the relationship between amyloid-beta(Aβ)accumulation and activation of the pyroptotic pathway in the retina.METHODS:The twelve 18-month-old TgAPPswePS1 transgenic mice and twelve 18-month-old wild-type C57BL/6J mice were used to investigate amyloid precursor protein(APP)and Aβexpression,retinal structural changes,and activation of pyroptosis in RGCs.Immunohistochemical analyses were performed to detect APP,Aβ,and pyroptosisrelated proteins[NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1,gasdermin D(GSDMD),interleukin(IL)-1β,and IL-18].Quantitative assessments of retinal nerve fiber layer(RNFL)thickness were conducted to evaluate retinal integrity.RESULTS:Compared to age-matched wild-type controls,TgAPPswePS1 transgenic mice exhibited significant upregulation of APP and Aβwithin RGCs.Histological analysis revealed reduced RNFL thickness,indicating structural degeneration.Notably,RGCs in transgenic mice showed robust immunoreactivity for NLRP3,caspase-1,and GSDMD,alongside elevated levels of IL-1βand IL-18,supporting the activation of pyroptosis.CONCLUSION:Aβaccumulation in RGCs is associated with retinal degeneration and activation of the pyroptosis pathway in aged TgAPPswePS1 mice.This study provides new insights into the inflammatory mechanisms underlying Aβ-related retinal neurodegeneration and suggests that targeting pyroptosis may represent a promising therapeutic strategy for retinal disorders linked to amyloid pathology.展开更多
基金Supported by the Science and Technology Program of Gansu Province,No.23JRRA1015.
文摘Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermin D(GSDMD)cleavage plays a pivotal role in disease progression through nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP-3)/caspase-1 classical and caspase-4/5/11 non-classical pathways.Targeting pyroptosis has emerged as a promising therapeutic strategy,with recent advances highlighting the potential of pyroptosis inhibitors such as small-molecule compounds,biologics,and repurposed drugs that specifically target NLRP3,caspases,or GSDMD to suppress inflammasome activation,block pore formation,and mitigate downstream inflammation.This review systematically summarizes the mechanisms and therapeutic effects of these inhibitors,while addressing critical challenges including drug specificity,delivery efficiency,and long-term safety,and explores their potential in combination therapies with existing IBD treatments to enhance clinical efficacy.By integrating preclinical and clinical evidence,we provide valuable insights into the translational prospects of pyroptosis-targeted therapies for precision management of IBD.
基金supported by the Natural Science Foundation of Fujian Province,No.2021J02035(to WX).
文摘Regulated cell death is a form of cell death that is actively controlled by biomolecules.Several studies have shown that regulated cell death plays a key role after spinal cord injury.Pyroptosis and ferroptosis are newly discovered types of regulated cell deaths that have been shown to exacerbate inflammation and lead to cell death in damaged spinal cords.Autophagy,a complex form of cell death that is interconnected with various regulated cell death mechanisms,has garnered significant attention in the study of spinal cord injury.This injury triggers not only cell death but also cellular survival responses.Multiple signaling pathways play pivotal roles in influencing the processes of both deterioration and repair in spinal cord injury by regulating pyroptosis,ferroptosis,and autophagy.Therefore,this review aims to comprehensively examine the mechanisms underlying regulated cell deaths,the signaling pathways that modulate these mechanisms,and the potential therapeutic targets for spinal cord injury.Our analysis suggests that targeting the common regulatory signaling pathways of different regulated cell deaths could be a promising strategy to promote cell survival and enhance the repair of spinal cord injury.Moreover,a holistic approach that incorporates multiple regulated cell deaths and their regulatory pathways presents a promising multi-target therapeutic strategy for the management of spinal cord injury.
基金supported by the Natural Science Fund of Fujian Province,No.2020J011058(to JK)the Project of Fujian Provincial Hospital for High-level Hospital Construction,No.2020HSJJ12(to JK)+1 种基金the Fujian Provincial Finance Department Special Fund,No.(2021)848(to FC)the Fujian Provincial Major Scientific and Technological Special Projects on Health,No.2022ZD01008(to FC).
文摘Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.
基金supported by the National Natural Science Foundation of China Regional Joint Innovation Fund (No.U22A20524)the Heilongjiang Province Natural Science Foundation Key projects (No.ZD2023C002).
文摘Bisphenol A(BPA)is an industrial pollutant that can cause immune impairment.Selenium acts as an antioxidant,as selenium deficiency often accompanies oxidative stress,resulting in organ damage.This study is the first to demonstrate that BPA and/or selenium deficiency induce pyroptosis and ferroptosis-mediated thymic injury in chicken and chicken lymphoma cell(MDCC-MSB-1)via oxidative stress-induced endoplasmic reticulum(ER)stress.We established a broiler chicken model of BPA and/or selenium deficiency exposure and collected thymus samples as research subjects after 42 days.The results demonstrated that BPA or selenium deficiency led to a decrease in antioxidant enzyme activities(T-AOC,CAT,and GSH-Px),accumulation of peroxides(H2O2 and MDA),significant upregulation of ER stress-relatedmarkers(GRP78,IER 1,PERK,EIF-2α,ATF4,and CHOP),a significant increase in iron ion levels,significant upregulation of pyroptosis-related gene(NLRP3,ASC,Caspase1,GSDMD,IL-18 and IL-1β),significantly increase ferroptosis-related genes(TFRC,COX2)and downregulate GPX4,HO-1,FTH,NADPH.In vitro experiments conducted in MDCC-MSB-1 cells confirmed the results,demonstrating that the addition of antioxidant(NAC),ER stress inhibitor(TUDCA)and pyroptosis inhibitor(Vx765)alleviated oxidative stress,endoplasmic reticulum stress,pyroptosis,and ferroptosis.Overall,this study concludes that the combined effects of oxidative stress and ER stress mediate pyroptosis and ferroptosis in chicken thymus induced by BPA exposure and selenium deficiency.
基金Sichuan Science and Technology Program(No.2021ZDZX0009).
文摘Background Oxidative stress significantly impacts growth performance and liver function in piglets.Ferulic acid(FA)works as an antioxidant,however,the role and mechanism of FA in the regulation of diquat-induced oxidative stress in piglets are less known.This study was designed to investigate the effects of FA on growth performance and antioxi-dant capacity in piglets with diquat challenge.Methods Thirty-two healthy DLY(Duroc×Landrace×Yorkshire)piglets(13.24±0.19 kg)were randomly divided into one of two diets including 0 or 4 g/kg FA for 14 d.On d 15,all pigs were intraperitoneally injected diquat or sterile saline.Results Dietary supplementation with ferulic acid(FA)significantly improved the average daily gain(ADG)and decreased feed-gain ratio(F/G)of piglets.Here,dietary FA supplementation reduced serum aspartate aminotrans-ferase(AST),alanine aminotransferase(ALT)activities in diquat challenged piglets.Furthermore,diquat infusion increased reactive oxygen radicals(ROS)level in liver,decreased the activities of total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC)and increased malondialdehyde(MDA)con-tent in the liver and serum.Supplementation with FA significantly increased T-AOC and T-SOD activities and decreased MDA and ROS levels.FA down-regulated gene and protein expression of Keap1,and up-regulated protein expression of Nrf2 and HO-1 in the liver of piglets with diquat challenge.Importantly,diquat challenge increased the ratio of late apoptosis,increased serum levels of IL-1β,IL-18 and lactate dehydrogenase(LDH),and up-regulated pyroptosis-related genes in the liver.FA supplementation reduced the ratio of late apoptosis and down-regulated mRNA expression of Caspase-1.Accordingly,FA addition reduced concentration of IL-1β,IL-18,and LDH under diquat challenge.Conclusions Diquat-induced oxidative stress reduced growth performance and impaired liver function in piglets.Dietary FA supplementation enhanced the antioxidant capacity and reduced the degree of hepatocyte pyroptosis,thereby alleviating the oxidative damage in the liver and mitigating the impact of diquat on growth performance of piglets.
文摘Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.
文摘The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.
基金supported by grants from the National Natural Science Foundation of China(8170060495 and 82170682)。
文摘Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.
基金supported by the National Natural Science Foundation of China(Grant No.82374382,82074361,82274293)school-level major project of Beijing University of Chinese Medicine(2023-JYB-JBZD-037)+1 种基金hospital-level project of Dongzhimen Hospital,Beijing University of Chinese Medicine(DZMG-XZYY-23002)Chinese Society of Traditional Chinese Medicine Practical Project(ZSL-003-02)。
文摘Background:The Shenlian formula demonstrates therapeutic bmodelenefits for diabetic kidney disease(DKD).Yet,its effectiveness in mitigating renal interstitial fibrosis(RIF)and the pharmacological underpinnings remain to be elucidated.This investigation seeks to delineate some of the formula’s potential mechanisms via experimental validation.Methods:The study initiated by inducing a DKD in rats through unilateral nephrectomy combined with streptozotocin(STZ)administration and an advanced glycation end products-bovine serum albumin(AGE-BSA)induced DKD model in HK-2 cells to assess the Shenlian formula’s renal protective effects.Renal tissues underwent pathological and immunohistochemical staining to evaluate improvements in RIF.Mechanistic insights were further obtained through techniques such as Western Blot,immunofluorescence,co-immunoprecipitation,enzyme-linked immunosorbent assay(ELISA),and protein docking analyses.Results:Shenlian formula could enhance renal function,alleviate tubular damage,suppress epithelial-mesenchymal transition(EMT),reduce extracellular matrix(ECM)deposition,and thus decelerate RIF progression.It notably decreased the expression of markers associated with pyroptosis(NOD-,LRR-and pyrin domain-containing protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),caspase1,cleaved-caspase1,gasdermin D(GSDMD),GSDMD-N,interleukin-18(IL-18),interleukin-1 beta(IL-1β)),effectively inhibiting renal cell pyroptosis.Moreover,the formula facilitated autophagy substrate(sequestosome 1(p62))degradation,efficiently restoring the autophagy pathway.It also modulated autophagy,impacting cell pyroptosis regulation.Conclusion:The Shenlian formula potentially inhibits cellular pyroptosis by modulating the autophagy pathway,thereby diminishing inflammation-induced renal injury and fibrosis.This finding suggests a novel therapeutic approach for managing DKD-induced fibrosis.
基金supported by grants from the National Natural Science Foundation of China (82160837)2019 Guangxi First-Class Discipline Construction Project (2019XK139)+3 种基金Qihuang Project High-level Talents Cultivation Program of Guangxi University of Chinese Medicine (2021007)2019 Traditional Chinese Medicine Inheritance and Innovation Talent Training Platform Construction Project (2019-41)Innovation Program of Guangxi Graduate Education of Guangxi University of Chinese Medicine (XYJ22024)2022 Guangxi (Young) Qihuang Scholar Training Program (2021-10)
文摘Background:Activation of NLRP3(NOD-,LRR-and pyrin domain-containing protein 3)inflammasomes induced by pyroptosis is crucial in metabolic dysfunction-associated steatohepatitis(MASH)progression.Adiponectin possesses an anti-inflammatory role in various liver diseases.This study aimed to evaluate the effects of adiponectin on MASH.Methods:Adiponectin-mediated anti-inflammatory mechanisms,effects on pyroptosis-related proteins,and activation of NLRP3 inflammasomes were investigated using methionine-choline-deficient(MCD)-induced MASH murine model and in vitro models.The degree of MASH inflammation in liver tissue of C57BL/6J mice was assessed using histopathology.Enzyme-linked immunosorbent assay was performed to measure levels of inflammatory factors[interleukin-18(IL-18),IL-1β,and tumor necrosis factor-α(TNF-α)]in mice serum and culture medium.Western blot and quantitative polymerase chain reaction were performed to analyze the expression of pyroptosis-related genes and proteins in liver tissues of mouse model and in vitro models.Macrophage recruitment in vitro was evaluated using co-culture of upper and lower chambers.Results:MASH developed in MCD diet mice[metabolic dysfunction-associated steatotic liver disease(MASLD)activity score=6]but not in methionine-choline-sufficient(MCS)diet mice(MASLD activity score=3).Compared to MCS-fed mice,MCD-fed mice showed increased serum levels of aspartate amino-transferase,IL-18,IL-1β,and TNF-αand higher MASLD activity score(P<0.001).Adiponectin inhibited these increases(P<0.05)and suppressed mRNA and protein levels of NLRP3,gasdermin-D(GSDMD),and GSDMD-N in liver tissues(P<0.05).In vitro,lipopolysaccharide(LPS)/palmitic acid(PA)increased the levels of IL-18,IL-1β,and TNF-α,mRNA expressions of CASP1 and GSDMD,and production of CASP1,NLRP3,GSDMD,and GSDMD-N(P<0.01).Adiponectin reduced the levels of these inflammatory fac-tors and downregulated the mRNA expression and protein generation of pyroptosis-related markers(P<0.05).HepG2 cells pretreated with LPS/PA recruited more J774A.1 cells(P<0.001)and increased inflam-matory factor secretion by J774A.1 cells(P<0.001).Adiponectin inhibited this recruitment and reduced inflammatory factor secretion(P<0.001).Conclusions:Adiponectin inhibits hepatocyte pyroptosis by reducing the production and activation of NLRP3 inflammasomes,CASP1,and GSDMD,thus improving the inflammatory response in MASH and possibly delaying or reversing MASLD progression.
文摘To the Editor:Pancreatic cancer is a malignancy characterized by a poor prog-nosis,with a 5-year survival rate of<10%[1].Furthermore,only a minority of patients(<20%)qualify for curative-intent resec-tion,and even among those who undergo this procedure,the risk of recurrence within three years remains alarmingly high,reach-ing up to 70%[2].Due to the lack of specific clinical manifes-tations of pancreatic cancer,most cases have metastasized or in-vaded the major vessels around the pancreas at the time of initial diagnosis,resulting in a low surgical resection rate.Even patients who undergo surgical resection often face a poor prognosis[3].In recent years,neoadjuvant chemotherapy using agents such as gemcitabine,5-fluorouracil,albumin-bound paclitaxel,modified fluorouracil/leucovorin plus irinotecan,and oxaliplatin(mFOLFIRI-NOX),targeted therapies addressing molecular subtypes of pan-creatic cancer,and immunotherapies targeting PD-1 and PD-L1 have shown efficacy in improving the overall prognosis of patients with pancreatic cancer,although the impact remains modest[4,5].Therefore,novel therapeutic strategies and prognostic evaluation systems are urgently needed to enhance the survival of patients with pancreatic cancer.
基金Supported by National Natural Science Foundation of China,No.82270863 and No.82470849Major Project of Anhui Provincial University Research Program,No.2023AH040400+1 种基金Joint Fund for Medical Artificial Intelligence,No.MAI2023Q026the Project of Health Commission Scientific Research in Anhui Province,No.AHWJ2024Aa20477.
文摘BACKGROUND Pyroptosis and ubiquitination have been identified as key processes influencing the pathogenesis of diabetes mellitus(DM).AIM To investigate the genes associated with the ubiquitin-proteasome system(UPS)and pyroptosis in type 2 DM(T2DM),and elucidate their mechanisms of action in T2DM.METHODS The datasets GSE76894,GSE41762,and GSE86469 were utilized in this study.UPS-related genes(UPSGs)and pyroptosis-related genes(PRGs)were obtained from existing literature.Differential expression analysis was performed to identify differentially expressed genes(DEGs).DEGs were intersected with UPSGs and PRGs to identify differentially expressed UPSGs and PRGs.Ubiquitin-pyroptosisrelated biomarkers were determined using Spearman’s correlation,t-tests,and receiver operating characteristic curve analysis.Pathway enrichment of biomarkers was assessed using Gene Set Enrichment Analysis(GSEA).Single sample GSEA(ssGSEA)and Spearman’s correlation were used to analyze the relationship between biomarkers and immune cells.A competitive endogenous RNA network was constructed.Subsequently,drugs related to the biomarkers were identified and a gene-drug network was established.In dataset GSE86469,single-cell sequencing was utilized to determine cell types.Finally,the expression levels of biomarkers were validated through quantitative PCR(qPCR)and western blot analysis.RESULTS A total of 581 DEGs were identified in GSE76894.Four genes[ATP binding cassette subfamily C member 8(ABCC8),retinol binding protein 4(RBP4),Ras protein-specific guanine nucleotide-releasing factor 1(RASGRF1),and solute carrier family 34 member 2(SLC34A2)]were identified as ubiquitin-pyroptosis-related biomarkers in T2DM,based on consistent expression trends and significant differences in GSE76894 and GSE41762.These biomarkers were enriched in oxidative phosphorylation and mitogen-activated protein kinase signaling pathways,which are relevant to DM.ssGSEA revealed significant differences in the enrichment scores of nine immune cell types between groups.A total of 17 microRNAs(miRNAs)and 36 long non-coding RNAs(lncRNAs)were identified,forming numerous miRNA-lncRNA interactions.Additionally,22 drugs related to the biomarkers,such as gliclazide and tretinoin,were identified.In GSE86469,eight cell types,including alpha and beta cells,were characterized.qPCR and western blot analysis confirmed that the expression trends of RASGRF1 and SLC34A2 were consistent with the findings in GSE76894.CONCLUSION This study identified four ubiquitin-pyroptosis-related biomarkers(ABCC8,RBP4,RASGRF1,and SLC34A2)in T2DM through bioinformatics analysis,providing novel insights into the diagnosis and treatment of T2DM.
基金supported by the Project of National Natural Science Foundation of China(Grant No.:82274219 and 81930117)the Key Project of Traditional Chinese Medicine Technology Development Plan of Jiangsu Province,China(Grant No.:ZD202201)Jiangsu Province Postgraduate Scientific Research Practice and Innovation Plan Project,China(Grant Nos.:KYCX21_1735 and SJCX21_0679).
文摘Sustained inflammatory responses are closely related to various severe diseases,and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment.Natural products have garnered considerable concern for the treatment of inflammation.Huanglian-Wumei decoction(HLWMD)is a classic prescription used for treating inflammatory diseases,but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated.Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory,we successfully obtained berberine(BBR)-chlorogenic acid(CGA)supramolecular(BCS),which is an herbal pair from HLWMD.Using a series of characterization methods,we confirmed the self-assembly mechanism of BCS.BBR and CGA were self-assembled and stacked into amphiphilic spherical supramolecules in a 2:1 molar ratio,driven by electrostatic interactions,hydrophobic interactions,andπ–πstacking;the hydrophilic fragments of CGA were outside,and the hydrophobic fragments of BBR were inside.This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules.Compared with free molecules,BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide(LPS)-induced pyroptosis.Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB(NF-κB)p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.
基金supported by the National Natural Science Foundation of China(Grant No.:82374552)Hunan Provincial Natural Science Foundation of China for Distinguished Young Scholars(Grant No.:2024JJ2086)+1 种基金the Science and Technology Innovation Program of Hunan Province,China(Grant No.:2022RC1220)Support Plan for High-level Health and Medical Talents in Hunan Province,China.
文摘Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative diseases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing.There are currently no effective strategies for slowing the progression of AD.Herein,we spotlight the dysregulation of lipid metabolism,particularly the elevation of ceramides(Cers),as a critical yet underexplored facet of AD pathogenesis.Our study delineates the role of Cers in promoting microglial pyroptosis,a form of programmed cell death distinct from apoptosis and necroptosis,characterized by cellular swelling,and membrane rupture mediated by the NLRP3 inflammasome pathway.Utilizing both in vivo experiments with amyloid precursor protein(APP)/presenilin 1(PS1)transgenic mice and in vitro assays with BV-2 microglial cells,we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin(ICA),a flavonoid with known antioxidant and anti-inflammatory properties.Our findings reveal a significant increase in Cers levels and pyroptosis markers(NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain,caspase-1,gasdermin D(GSDMD),and interleukin-18(IL-18))in the brains of AD model mice,indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis.Conversely,ICA treatment effectively reduces these pyroptotic markers and Cer levels,thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD.This study not only advances our understanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy,capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2(COX-2)-NLRP3 inflammasome-gasdermin D(GSDMD)axis.Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of targeting microglial pyroptosis in AD.
基金supported by the National Natural Science Foundation of China(82374367)Jiangxi Provincial Natural Science Foundation(20242BAB26163,20232BAB206144)+4 种基金Jiangxi Province Key Laboratory of Traditional Chinese Medicine for Cardiovascular Diseases(20242BCC32096)NATCM’s Project of High-level Construction of Key TCM Disciplines(zyyzdxk-2023113)Project of Key Discipline Construction Fund of Jiangxi University of Chinese Medicine(2023jzzdxk032)Science and Technology Innovation Team Development Program of Jiangxi University of Chinese Medicine(CXTD22011)National Traditional Chinese Medicine Inheritance and Innovation Center Construction Project.
文摘Background:Atherosclerosis(AS),the primary pathological foundation of cardiovascular diseases,is characterized by intricate processes including inflammation,lipid metabolism disorders,and pyroptosis.While the traditional Chinese medicine compound Dingxin Recipe(DXR)has demonstrated definitive clinical efficacy in treating AS,its therapeutic mechanisms remain unclear.This study employed an integrated approach combining network pharmacology,molecular docking,and molecular dynamics simulations(MDS)to investigate DXR’s anti-AS mechanisms.Methods:Active ingredients and targets of DXR were identified and screened using databases such as GeneCards,OMIM,and TCMSP.An“ingredient-target-disease”network was constructed to visualize these interactions.Molecular docking was utilized to assess the binding affinity between key ingredients and their respective targets.Additionally,MDS were conducted to analyze the stability of these complexes,providing robust evidence for further clinical applications and in-depth research.Results:Through network pharmacology analysis,we identified 99 active drug components,934 gene targets,and 1463 disease targets associated with DXR.Protein-protein interaction analysis revealed central regulatory nodes.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that these components primarily modulate processes such as inflammatory response and transcription factor activation,and are closely linked to the AGERAGE signaling pathway,lipid metabolism,and atherosclerosis pathways.Molecular docking confirmed strong binding potential between the components and their targets,while MDS further validated the stability of these interactions.Conclusion:This study elucidates that the active ingredients in DXR alleviate AS by mitigating inflammatory responses and inhibiting pyroptosis through the suppression of inflammatory factor release.These findings provide a scientific foundation for the clinical application of DXR in AS treatment.
基金supported by grants from the National Key Research and Development Program of China(2023YFA1801804,2022YFA0807000,2021YFA1101300,2021YFA1101800,and 2020YFA0112503)the National Natural Science Foundation of China(82171149,82371166,81970892,82330033,82030029,92149304,82071053,and 82171144)+5 种基金the Shenzhen Science and Technology Program(JCYJ20230807114700001,JCYJ20210324125608022,and JCYJ20190814093401920)the Guangdong Basic and Applied Basic Research Foundation(2024A1515010548)the Science and Technology Department of Sichuan Province(2021YFS0371)the Jiangsu Provincial Scientific Research Center of Applied Mathematics(BK20233002)the Open Research Fund of Guangdong Academy of Medical Sciences(YKY‐KF202201)the Taishan Scholars Project-Young Experts Program of Shandong Province(tsqn202211357).
文摘Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiological factors have been shown to cause acquired SNHL,such as ototoxic drugs,noise exposure,aging,infections,and diseases.Several programmed cell death(PCD)pathways have been reported to be involved in SNHL,especially some novel PCD pathways that have only recently been reported,such as ferroptosis,necroptosis,and pyroptosis.Here we summarize these PCD pathways and their roles and mechanisms in SNHL,aiming to provide new insights and potential therapeutic strategies for SNHL by targeting these PCD pathways.
基金supported by a fund from the Hainan Provincial Natural Science Foundation of China(No.821MS156).
文摘Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI.However,the mechanism remains unclear.This study aimed to investigate whether c-Myc affects CIRI by regulating Serine/Arginine-rich Splicing Factor 1(SRSF1)/NLRP1-mediated pyroptosis.Methods:Oxygen-glucose deprivation/reperfusion(OGD/R)induced neuroblastoma cells for the establishment of an in vitro CIRI model.The levels of c-Myc and SRSF1,cell viability,the expression of pyroptosis-related factors,and the interaction between SRSF1 and NLRP1 were evaluated.Results:The expression of c-Myc and SRSF1 was decreased in OGD/R-induced neuroblastoma cells.c-Myc overexpression increased c-Myc and SRSF1 expression and cell viability in OGD/Rinduced neuroblastoma cells while inhibiting NLRP1,Caspase1,apoptosis-associated speck-like protein containing a CARD(ASC),interleukin-1beta(IL-1β),IL-18,and lactate dehydrogenase levels and pyroptosis.C-Myc was positively correlated with SRSF1.SRSF1 low expression reversed the effects of c-Myc on the above indicators in OGD/Rinduced neuroblastoma cells.Mechanically,SRSF1 interacted with NLRP1.SRSF1 was negatively correlated with NLRP1.The NLRP1 activator muramyl dipeptide(MDP)reversed the SRSF1 effect on OGD/R-induced neuroblastoma cells.Conclusion:Our results indicated that c-Myc reduced OGD/R-induced neuroblastoma cell pyroptosis by inhibiting NLRP1 activation by positive feedback SRSF1 signal.Our findings suggested that the c-Myc/SRSF1 axis might be a new strategy for treating CIRI in the clinic.
基金supported by Chung Shan Medical University and Changhua Christian Hospital(CSMU-CCH-111-08)supported by research grants from the Chung Shan Medical University Hospital,Taichung,Taiwan(CSH-2023-C-023)the National Science and Technology Council,Taiwan,for financially supporting this research under Contract Nos.NSTC 112-2320-B-040-017,NSTC112-2314-B-040-009,and NSTC 112-2320-B-040-011.
文摘Background:Acute respiratory distress syndrome(ARDS)is the major therapeutic dilemma associated with significant inflammation and severe pulmonary dysfunction.Liriodendrin is a bioactive compound extract from traditional Chinesemedicine,historically utilized formodulating inflammatory responses and alleviating symptoms in multiple diseasemodels.Methods:At present,BALB/c mice to explore the effects of liriodendrin on lipopolysaccharide(LPS)-induced ARDS.Before LPS was administered,the mice were treated with either liriodendrin or dexamethasone.Leukocyte infiltration,lung edema,and alveolar-capillary barrier integrity were evaluated in the bronchoalveolar lavage fluid(BALF)and pulmonary parenchyma.The expression of adhesion molecules and proinflammatory cytokines in BALF was evaluated by enzyme-linked immunosorbent assay.Western blotting assay facilitated the analysis of the expression or phosphorylation of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),NOD-like receptor family pyrin domain-containing 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),cleaved caspase-1(CL-csapase-1),nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB),inhibitor of kappa B(IκB),mitogen-activated protein kinase(MAPK),and protein kinase B(Akt)in the lungs.In addition,the anti-inflammatory effects of liriodendrin were evaluated in LPS-stimulated RAW264.7 macrophages.Before LPS was administered,the RAW264.7 macrophages were treated with either liriodendrin or dexamethasone.Nitric Oxide(NO)production was measured using the Griess reaction assay,while ELISA assessed IL-1β,IL-6,and TNF-αlevels.Western blot analysis evaluated NF-κB phosphorylation and the expression of NLRP3,ASC,and CLcaspase-1.Results:These outcomes revealed that liriodendrin intervention markedly ameliorated the pathological features of LPS-induced ARDS,including leukocyte infiltration,lung edema,and alveolar-capillary barrier disruption.Liriodendrin also reduced the LPS-induced secretion of intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesionmolecule-1(VCAM-1),expression of iNOS and COX2,and production of proinflammatory cytokines.Finally,we further discovered that the concentration trend of liriodendron amelioration ofARDSwas similar to those ofNLRP3 formation,NF-κB pathway activation,and p38 MAPK,c-Jun N-terminal kinase(JNK),and Akt phosphorylation but not to that of extracellular signal-regulated kinase(ERK)phosphorylation.Liriodendrin inhibited LPS-induced inflammatory responses in RAW264.7 macrophages.It markedly reduced NO production,propro-inflammatorytokines,NF-κB phosphorylation,and NLRP3 formation.Conclusions:In summary,liriodendrin effectively ameliorated the pathological features of LPS-induced ARDS inmice,demonstrating significant anti-inflammatory properties attributed to NLRP3 formation through NF-κB pathway activation by p38MAPK,JNK,and Akt phosphorylation.In LPS-treated RAW264.7 macrophages,liriodendrin reduced NO production,pro-inflammatory cytokines,and NLRP3 formation,suggesting its potential as an agent for ARDS and relative inflammation.
基金supported by the National Natural Science Foundation of China(32373062)the Natural Science Foundation of Shandong Province(ZR2023MC144)Funds of Shandong Province Modern Agricultural Technology System Innovation Team Program(SDAIT-21-10).
文摘Background Deoxynivalenol(DON)is a mycotoxin that severely pollutes feed ingredients,and methods for reducing DON toxicity have become a significant research direction.Chlorogenic acid(CGA)is an active polyphenol found in some plants,which has anti-inflammatory and antioxidant properties and a protective effect on animal intestinal health.The effects of CGA on DON-induced pyroptosis in the intestinal porcine epithelial cell line-J2(IPEC-J2)and its potential mechanism were explored in this study.Results IPEC-J2 cells viability and membrane integrity were inversely correlated with DON concentration.Compared to those in the group treated with DON alone at 2,500 ng/mL,pretreatment with 80μmol/L CGA for 4 h significantly improved cell viability(P<0.01),and the alleviation of typical pyroptotic symptoms induced by DON were observed,including reduced cellular DNA fragmentation,decreased release of lactate dehydrogenase(LDH),normalized ROS levels,restoration of extracellularCa2+andK+contents to normal levels(P<0.01),as well as suppressed the enzyme activities of caspase-1 and caspase-4(P<0.01).Additionally,the mRNA expression levels of TNF,MDP,NOD2,TLR4,ASC and GSDMD were significantly improved(P<0.01),while both mRNA and protein expression levels of NF-κB,NLRP3,caspase-1,IL-1βand IL-18 were significantly upregulated(P<0.01)in the CGA+DON group,compare to those in the DON group.Conclusion Pretreatment with 80μmol/L CGA for 4 h effectively alleviated pyroptosis in IPEC-J2 cells induced by 2,500 ng/mL of DON through inhibiting activation of the NF-κB/NLRP3/capase-1 pathway.
基金Supported by Natural Science Foundation of Zhejiang Province(No.LY18H120009)Wenzhou Basic Scientific Research Project(No.2025K0279)+1 种基金Xi’an Health and Wellness Committee General Cultivation Project(No.2023ms08)Shaanxi Province Health and Health High-Level Talents(Team)Training Program Young Talents Project.
文摘AIM:To investigate whether pyroptosis contributes to retinal ganglion cell(RGC)degeneration in aged TgAPPswePS1 transgenic mice and to explore the relationship between amyloid-beta(Aβ)accumulation and activation of the pyroptotic pathway in the retina.METHODS:The twelve 18-month-old TgAPPswePS1 transgenic mice and twelve 18-month-old wild-type C57BL/6J mice were used to investigate amyloid precursor protein(APP)and Aβexpression,retinal structural changes,and activation of pyroptosis in RGCs.Immunohistochemical analyses were performed to detect APP,Aβ,and pyroptosisrelated proteins[NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1,gasdermin D(GSDMD),interleukin(IL)-1β,and IL-18].Quantitative assessments of retinal nerve fiber layer(RNFL)thickness were conducted to evaluate retinal integrity.RESULTS:Compared to age-matched wild-type controls,TgAPPswePS1 transgenic mice exhibited significant upregulation of APP and Aβwithin RGCs.Histological analysis revealed reduced RNFL thickness,indicating structural degeneration.Notably,RGCs in transgenic mice showed robust immunoreactivity for NLRP3,caspase-1,and GSDMD,alongside elevated levels of IL-1βand IL-18,supporting the activation of pyroptosis.CONCLUSION:Aβaccumulation in RGCs is associated with retinal degeneration and activation of the pyroptosis pathway in aged TgAPPswePS1 mice.This study provides new insights into the inflammatory mechanisms underlying Aβ-related retinal neurodegeneration and suggests that targeting pyroptosis may represent a promising therapeutic strategy for retinal disorders linked to amyloid pathology.
