Inflammation plays a key role in driving the secondary brain injury that follows ischemic stroke.Melatonin is an endogenous neuroendocrine hormone that regulates mitochondrial homeostasis.However,the role and mechanis...Inflammation plays a key role in driving the secondary brain injury that follows ischemic stroke.Melatonin is an endogenous neuroendocrine hormone that regulates mitochondrial homeostasis.However,the role and mechanisms by which melatonin regulates microglial pyroptosis and the inflammatory cascade through double-stranded DNA(dsDNA)-sensing cyclic GMP-AMP synthase(cGAS)signaling warrant further study.Using middle cerebral artery occlusion mice,we investigated the effects of melatonin on cGAS-mediated pyroptosis and neuroinflammation.Middle cerebral artery occlusion model mice exhibited significantly increased DNA damage and cytoplasmic dsDNA release,as reflected byγH2AX staining,as well as heightened activation of the cytosolic dsDNA-sensing cGAS-STING pathway,both of which were notably suppressed by melatonin treatment.Melatonin also mitigated NOD-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome activation and nuclear factor(NF)-κB/gasdermin D-mediated pyroptosis in microglia following ischemic stroke,while exhibiting the capacity to attenuate the immune response to ischemia in mice.This led to reduced infiltration of peripheral neutrophils and monocytes/macrophages in the ischemic brain.Specifically,melatonin administration resulted in reductions in the numbers of ionized calcium-binding adapter molecule 1-positive cells and production of interleukin-6 and tumor necrosis factor-αby microglia.Regarding neurological outcomes,melatonin significantly reduced cerebral infarct volume and ameliorated neurological deficits in mice.Notably,the neuroprotective effect of melatonin was correlated with the inhibition of cGAS activity.We also developed and tested melatonin co-loaded macrophage membrane-biomimetic reactive oxygen species-responsive nanoparticles(Mф-MLT@FNGs),which exhibited therapeutic properties in middle cerebral artery occlusion mice.Our findings suggest that melatonin acts on microglial pyroptosis to inhibit neuroinflammation and reshape the immune microenvironment through regulation of the cGAS-STING-NF-κB signaling pathway.By doing so,melatonin rescues damaged brain tissue and protects neurological function,highlighting its potential as a neuroprotective treatment for ischemic stroke.展开更多
Colorectal cancer(CRC)remains a major global health challenge,with high recurrence and mortality despite advances in surgery,chemotherapy,and immunotherapy.The study by He et al identifies a novel mechanism by which p...Colorectal cancer(CRC)remains a major global health challenge,with high recurrence and mortality despite advances in surgery,chemotherapy,and immunotherapy.The study by He et al identifies a novel mechanism by which peroxiredoxin 1(Prdx1)inhibits CRC progression through induction of pyroptosis,a pro-inflammatory form of programmed cell death.Traditionally viewed as an intracellular antioxidant that protects tumors from oxidative stress,Prdx1 assu-mes a paradoxical immunogenic role when released extracellularly as a damageassociated molecular pattern.Using patient samples,recombinant protein assays,and murine xenograft models,the authors demonstrate that Prdx1 activates the NOD-,LRR-and pyrin domain-containing protein 3 inflammasome/caspase-1/gasdermin D pathway,triggering membrane pore formation,tumor cell lysis,and release of interleukin-1β/interleukin-18.This cascade not only halts tumor proliferation,invasion,and migration but may also enhance anti-tumor immune surveillance.The study’s strengths include rigorous mechanistic validation,clinical cohort data,inhibitor-based causal proof,and in vivo confirmation.However,questions remain regarding the upstream receptor for Prdx1,heterogeneity across CRC subtypes,and the balance between therapeutic benefit and inflammatory toxicity.By establishing Prdx1-induced pyroptosis as a driver of tumor suppression,this work advances a promising paradigm in CRC therapy,linking cell death to immune activation and pointing toward future biomarker-driven,pyroptosis-based interventions.展开更多
Phosphodiesterase 4 is a key enzyme involved in the regulation of cell signal transduction,but its role in subarachnoid hemorrhage remains unclear.Neuronal pyroptosis has been reported to be involved in early brain in...Phosphodiesterase 4 is a key enzyme involved in the regulation of cell signal transduction,but its role in subarachnoid hemorrhage remains unclear.Neuronal pyroptosis has been reported to be involved in early brain injury after subarachnoid hemorrhage.This study aimed to investigate whether phosphodiesterase 4 contributes to early brain injury after subarachnoid hemorrhage by mediating neuronal pyroptosis and its related mechanisms.Endovascular perforation of male C57BL/6J mice was performed to model subarachnoid hemorrhage in vivo,and oxyhemoglobin was added to the culture medium of primary neurons to model subarachnoid hemorrhage in vitro.A phosphodiesterase 4-specific inhibitor,etazolate,was intraperitoneally injected 30 minutes after subarachnoid hemorrhage induction.Small interfering RNA(siRNA)was administered intracerebroventricularly 72 hours before subarachnoid hemorrhage to achieve genetic knockdown of phosphodiesterase 4.To investigate the mechanism,a nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3(NLRP3)-specific agonist,nigericin,was intracerebroventricularly injected 60 minutes before subarachnoid hemorrhage.Neuronal phosphodiesterase 4 expression increased after subarachnoid hemorrhage and reached the highest point at 24 hours.Etazolate treatment reduced neurological deficits and brain edema in mice,alleviated neuronal pyroptosis and inflammatory response,and improved neuronal injury.Treatment with phosphodiesterase 4 siRNA had the same neuroprotective effects as etazolate.Mechanistically,phosphodiesterase 4 triggered the nuclear factor kappa-B pathway,and simultaneously caused lysosomal and mitochondrial dysfunction after subarachnoid hemorrhage,which promoted NLRP3 inflammasome activation and induced neuronal pyroptosis.Blocking of phosphodiesterase 4 inhibited the nuclear factor kappa-B pathway,and improved lysosome and mitochondrial function.Activation of NLRP3 reversed the neuroprotective effects of etazolate without affecting phosphodiesterase 4 expression.Together,the results indicate that phosphodiesterase 4 regulates NLRP3-mediated neuronal pyroptosis in early brain injury after subarachnoid hemorrhage.Phosphodiesterase 4 may be a potential therapeutic molecular target for subarachnoid hemorrhage.展开更多
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
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 The pathophysiology of diabetic kidney disease(DKD)is complex.Interfering with the processes of pyroptosis and fibrosis is an effective strategy for slowing DKD progression.Previous studies have revealed th...BACKGROUND The pathophysiology of diabetic kidney disease(DKD)is complex.Interfering with the processes of pyroptosis and fibrosis is an effective strategy for slowing DKD progression.Previous studies have revealed that nuclear receptor subfamily 4 group A member 1(NR4A1)may serve as a novel pathogenic element in DKD;however,the specific mechanism by which it contributes to pyroptosis and fibrosis in DKD is unknown.AIM To investigate the role of NR4A1 in renal pyroptosis and fibrosis in DKD and possible molecular mechanisms.METHODS Streptozotocin 60 mg/kg was injected intraperitoneally to establish a rat model of DKD.Typically,45 mmol/L glucose[high glucose(HG)]was used to activate HK-2 cells to mimic the DKD model in vitro.HK-2 cells were transfected with NR4A1 siRNA to silence NR4A1.RESULTS NR4A1 was elevated in renal tissues of DKD rats and HG-stimulated HK-2 cells.Concurrently,NOD-like receptor protein 3(NLRP3)and phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathways were triggered,and pyroptosis and expression of fibrosis-linked elements was increased in vivo and in vitro.These alterations were significantly reversed via NR4A1 silencing.CONCLUSION Inhibition of NR4A1 mitigated pyroptosis and fibrosis via suppressing NLRP3 activation and the PI3K/AKT pathway in HG-activated HK-2 cells.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
Microglial pyroptosis and neuroinflammation have been implicated in the pathogenesis of sepsis-associated encephalopathy(SAE).OGT-mediated O-GlcNAcylation is involved in neurodevelopment and injury.However,its regulat...Microglial pyroptosis and neuroinflammation have been implicated in the pathogenesis of sepsis-associated encephalopathy(SAE).OGT-mediated O-GlcNAcylation is involved in neurodevelopment and injury.However,its regulatory function in microglial pyroptosis and involvement in SAE remains unclear.In this study,we demonstrated that OGT deficiency augmented microglial pyroptosis and exacerbated secondary neuronal injury.Furthermore,OGT inhibition impaired cognitive function in healthy mice and accelerated the progression in SAE mice.Mechanistically,OGT-mediated O-GlcNAcylation of ATF2 at Ser44 inhibited its phosphorylation and nuclear translocation,thereby amplifying NLRP3 inflammasome activation and promoting inflammatory cytokine production in microglia in response to LPS/Nigericin stimulation.In conclusion,this study uncovers the critical role of OGT-mediated O-GlcNAcylation in modulating microglial activity through the regulation of ATF2 and thus protects against SAE progression.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: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
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 Inflammatory bowel disease(IBD)is closely associated with intestinal microbiota dysbiosis and metabolic dysfunction.The aim of this study was to explore the protective effects and mechanisms of the probioti...Background Inflammatory bowel disease(IBD)is closely associated with intestinal microbiota dysbiosis and metabolic dysfunction.The aim of this study was to explore the protective effects and mechanisms of the probiotic Bacillus velezensis MZ09,which produces branched-chain short-chain fatty acids(BSCFAs),against the dextran sulfate sodium(DSS)-induced colitis in piglets.Results In this study,a DSS-induced piglet colitis model was established to explore the impact of MZ09.Pretreatment with MZ09 significantly alleviated the symptoms of colitis in piglets.For example,the disease activity index(DAI)score decreased,the length of the colon was restored,and splenomegaly was alleviated.MZ09 enhanced intestinal barrier integrity by upregulating the expression of tight junction proteins such as Claudin-1,Occludin,and ZO-1.Using 16S rRNA analysis,we found that MZ09 could remodel the intestinal microbiota.MZ09 increased the abundance of beneficial bacteria such as Firmicutes and Lactobacillus while suppressing the growth of harmful bacteria such as Proteobacteria and Escherichia-Shigella.MZ09 also increased the levels of short-chain fatty acids(SCFAs)in the colon.The increased SCFA content activated G-protein-coupled receptor 43(GPR43),which increased the phosphorylation of signal transducer and activator of transcription 3(STAT3)and promoted the production of the antiinflammatory cytokine interleukin-10(IL-10).Mechanistically,MZ09 mitigated mitochondrial damage via the STAT3/hypoxia-inducible factor 1α(HIF-1α)axis.This action inhibits nucleotide-binding oligomerization domain,leucinerich repeat and pyrin domain-containing 3(NLRP3)inflammasome-mediated pyroptosis,thus reducing the release of the proinflammatory cytokines IL-1βand IL-18.Conclusions B.velezensis MZ09 alleviates DSS-induced colitis in piglets through multiple pathways,including gut microbiota remodeling,SCFAs–GPR43–STAT3 axis activation,and NLRP3 inflammasome-mediated pyroptosis suppression.These findings provide a new theoretical basis for the development of targeted intervention strategies for IBD,suggesting that MZ09 represents a potentially promising therapeutic agent for IBD treatment.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
基金supported by the Natural Science Foundation of Heilongjiang Province,No.YQ2021H011(to QL)China Postdoctoral Science Foundation,Nos.2020M670925,2022T150172(to QL)+2 种基金Postdoctoral Foundation of Heilongjiang Province,Nos.LBH‐Z19027,LBH‐TZ2019(to QL)Institute Cultivation Fund,No.PYMS2023-1(to QL)Natural Science Foundation of Jiangsu Province,No.BK20241233(to YL).
文摘Inflammation plays a key role in driving the secondary brain injury that follows ischemic stroke.Melatonin is an endogenous neuroendocrine hormone that regulates mitochondrial homeostasis.However,the role and mechanisms by which melatonin regulates microglial pyroptosis and the inflammatory cascade through double-stranded DNA(dsDNA)-sensing cyclic GMP-AMP synthase(cGAS)signaling warrant further study.Using middle cerebral artery occlusion mice,we investigated the effects of melatonin on cGAS-mediated pyroptosis and neuroinflammation.Middle cerebral artery occlusion model mice exhibited significantly increased DNA damage and cytoplasmic dsDNA release,as reflected byγH2AX staining,as well as heightened activation of the cytosolic dsDNA-sensing cGAS-STING pathway,both of which were notably suppressed by melatonin treatment.Melatonin also mitigated NOD-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome activation and nuclear factor(NF)-κB/gasdermin D-mediated pyroptosis in microglia following ischemic stroke,while exhibiting the capacity to attenuate the immune response to ischemia in mice.This led to reduced infiltration of peripheral neutrophils and monocytes/macrophages in the ischemic brain.Specifically,melatonin administration resulted in reductions in the numbers of ionized calcium-binding adapter molecule 1-positive cells and production of interleukin-6 and tumor necrosis factor-αby microglia.Regarding neurological outcomes,melatonin significantly reduced cerebral infarct volume and ameliorated neurological deficits in mice.Notably,the neuroprotective effect of melatonin was correlated with the inhibition of cGAS activity.We also developed and tested melatonin co-loaded macrophage membrane-biomimetic reactive oxygen species-responsive nanoparticles(Mф-MLT@FNGs),which exhibited therapeutic properties in middle cerebral artery occlusion mice.Our findings suggest that melatonin acts on microglial pyroptosis to inhibit neuroinflammation and reshape the immune microenvironment through regulation of the cGAS-STING-NF-κB signaling pathway.By doing so,melatonin rescues damaged brain tissue and protects neurological function,highlighting its potential as a neuroprotective treatment for ischemic stroke.
文摘Colorectal cancer(CRC)remains a major global health challenge,with high recurrence and mortality despite advances in surgery,chemotherapy,and immunotherapy.The study by He et al identifies a novel mechanism by which peroxiredoxin 1(Prdx1)inhibits CRC progression through induction of pyroptosis,a pro-inflammatory form of programmed cell death.Traditionally viewed as an intracellular antioxidant that protects tumors from oxidative stress,Prdx1 assu-mes a paradoxical immunogenic role when released extracellularly as a damageassociated molecular pattern.Using patient samples,recombinant protein assays,and murine xenograft models,the authors demonstrate that Prdx1 activates the NOD-,LRR-and pyrin domain-containing protein 3 inflammasome/caspase-1/gasdermin D pathway,triggering membrane pore formation,tumor cell lysis,and release of interleukin-1β/interleukin-18.This cascade not only halts tumor proliferation,invasion,and migration but may also enhance anti-tumor immune surveillance.The study’s strengths include rigorous mechanistic validation,clinical cohort data,inhibitor-based causal proof,and in vivo confirmation.However,questions remain regarding the upstream receptor for Prdx1,heterogeneity across CRC subtypes,and the balance between therapeutic benefit and inflammatory toxicity.By establishing Prdx1-induced pyroptosis as a driver of tumor suppression,this work advances a promising paradigm in CRC therapy,linking cell death to immune activation and pointing toward future biomarker-driven,pyroptosis-based interventions.
基金supported by the National Natural Science Foundation of China,No.81870927(to ZH)the Natural Science Foundation Project ofChongqing Science and Technology Commission,No.CSTB2023NSCQ-MSX0112(to ZH).
文摘Phosphodiesterase 4 is a key enzyme involved in the regulation of cell signal transduction,but its role in subarachnoid hemorrhage remains unclear.Neuronal pyroptosis has been reported to be involved in early brain injury after subarachnoid hemorrhage.This study aimed to investigate whether phosphodiesterase 4 contributes to early brain injury after subarachnoid hemorrhage by mediating neuronal pyroptosis and its related mechanisms.Endovascular perforation of male C57BL/6J mice was performed to model subarachnoid hemorrhage in vivo,and oxyhemoglobin was added to the culture medium of primary neurons to model subarachnoid hemorrhage in vitro.A phosphodiesterase 4-specific inhibitor,etazolate,was intraperitoneally injected 30 minutes after subarachnoid hemorrhage induction.Small interfering RNA(siRNA)was administered intracerebroventricularly 72 hours before subarachnoid hemorrhage to achieve genetic knockdown of phosphodiesterase 4.To investigate the mechanism,a nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3(NLRP3)-specific agonist,nigericin,was intracerebroventricularly injected 60 minutes before subarachnoid hemorrhage.Neuronal phosphodiesterase 4 expression increased after subarachnoid hemorrhage and reached the highest point at 24 hours.Etazolate treatment reduced neurological deficits and brain edema in mice,alleviated neuronal pyroptosis and inflammatory response,and improved neuronal injury.Treatment with phosphodiesterase 4 siRNA had the same neuroprotective effects as etazolate.Mechanistically,phosphodiesterase 4 triggered the nuclear factor kappa-B pathway,and simultaneously caused lysosomal and mitochondrial dysfunction after subarachnoid hemorrhage,which promoted NLRP3 inflammasome activation and induced neuronal pyroptosis.Blocking of phosphodiesterase 4 inhibited the nuclear factor kappa-B pathway,and improved lysosome and mitochondrial function.Activation of NLRP3 reversed the neuroprotective effects of etazolate without affecting phosphodiesterase 4 expression.Together,the results indicate that phosphodiesterase 4 regulates NLRP3-mediated neuronal pyroptosis in early brain injury after subarachnoid hemorrhage.Phosphodiesterase 4 may be a potential therapeutic molecular target for subarachnoid hemorrhage.
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 Research Fund for Academician Lin He New Medicine,No.JYHL2022FMS02.
文摘BACKGROUND The pathophysiology of diabetic kidney disease(DKD)is complex.Interfering with the processes of pyroptosis and fibrosis is an effective strategy for slowing DKD progression.Previous studies have revealed that nuclear receptor subfamily 4 group A member 1(NR4A1)may serve as a novel pathogenic element in DKD;however,the specific mechanism by which it contributes to pyroptosis and fibrosis in DKD is unknown.AIM To investigate the role of NR4A1 in renal pyroptosis and fibrosis in DKD and possible molecular mechanisms.METHODS Streptozotocin 60 mg/kg was injected intraperitoneally to establish a rat model of DKD.Typically,45 mmol/L glucose[high glucose(HG)]was used to activate HK-2 cells to mimic the DKD model in vitro.HK-2 cells were transfected with NR4A1 siRNA to silence NR4A1.RESULTS NR4A1 was elevated in renal tissues of DKD rats and HG-stimulated HK-2 cells.Concurrently,NOD-like receptor protein 3(NLRP3)and phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathways were triggered,and pyroptosis and expression of fibrosis-linked elements was increased in vivo and in vitro.These alterations were significantly reversed via NR4A1 silencing.CONCLUSION Inhibition of NR4A1 mitigated pyroptosis and fibrosis via suppressing NLRP3 activation and the PI3K/AKT pathway in HG-activated HK-2 cells.
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.
基金supported by the Jiangsu Provincial Medical Key Discipline(Laboratory)Cultivation Unit(JSDW202249)the Natural Science Foundation of Jiangsu Province(BK20211108)+4 种基金a Scientific Research Project of the Health Commission of Nantong(MS2023035)Nantong Natural Science Foundation(JC2023114)the Scientific Research Innovation Team of Kangda College of Nanjing Medical University(KD2022KYCXTD005)Nantong University Clinical Medicine Special Project(2022JY005)the Postgraduate Research&Practice Innovation Program of Jiangsu province(KYCX23_3416).
文摘Microglial pyroptosis and neuroinflammation have been implicated in the pathogenesis of sepsis-associated encephalopathy(SAE).OGT-mediated O-GlcNAcylation is involved in neurodevelopment and injury.However,its regulatory function in microglial pyroptosis and involvement in SAE remains unclear.In this study,we demonstrated that OGT deficiency augmented microglial pyroptosis and exacerbated secondary neuronal injury.Furthermore,OGT inhibition impaired cognitive function in healthy mice and accelerated the progression in SAE mice.Mechanistically,OGT-mediated O-GlcNAcylation of ATF2 at Ser44 inhibited its phosphorylation and nuclear translocation,thereby amplifying NLRP3 inflammasome activation and promoting inflammatory cytokine production in microglia in response to LPS/Nigericin stimulation.In conclusion,this study uncovers the critical role of OGT-mediated O-GlcNAcylation in modulating microglial activity through the regulation of ATF2 and thus protects against SAE progression.
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 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.
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 the National Natural Science Foundation of China(32372924)the Major Industrialisation of Scientific and Technological Achievements in Heilongjiang Province(CG24019).
文摘Background Inflammatory bowel disease(IBD)is closely associated with intestinal microbiota dysbiosis and metabolic dysfunction.The aim of this study was to explore the protective effects and mechanisms of the probiotic Bacillus velezensis MZ09,which produces branched-chain short-chain fatty acids(BSCFAs),against the dextran sulfate sodium(DSS)-induced colitis in piglets.Results In this study,a DSS-induced piglet colitis model was established to explore the impact of MZ09.Pretreatment with MZ09 significantly alleviated the symptoms of colitis in piglets.For example,the disease activity index(DAI)score decreased,the length of the colon was restored,and splenomegaly was alleviated.MZ09 enhanced intestinal barrier integrity by upregulating the expression of tight junction proteins such as Claudin-1,Occludin,and ZO-1.Using 16S rRNA analysis,we found that MZ09 could remodel the intestinal microbiota.MZ09 increased the abundance of beneficial bacteria such as Firmicutes and Lactobacillus while suppressing the growth of harmful bacteria such as Proteobacteria and Escherichia-Shigella.MZ09 also increased the levels of short-chain fatty acids(SCFAs)in the colon.The increased SCFA content activated G-protein-coupled receptor 43(GPR43),which increased the phosphorylation of signal transducer and activator of transcription 3(STAT3)and promoted the production of the antiinflammatory cytokine interleukin-10(IL-10).Mechanistically,MZ09 mitigated mitochondrial damage via the STAT3/hypoxia-inducible factor 1α(HIF-1α)axis.This action inhibits nucleotide-binding oligomerization domain,leucinerich repeat and pyrin domain-containing 3(NLRP3)inflammasome-mediated pyroptosis,thus reducing the release of the proinflammatory cytokines IL-1βand IL-18.Conclusions B.velezensis MZ09 alleviates DSS-induced colitis in piglets through multiple pathways,including gut microbiota remodeling,SCFAs–GPR43–STAT3 axis activation,and NLRP3 inflammasome-mediated pyroptosis suppression.These findings provide a new theoretical basis for the development of targeted intervention strategies for IBD,suggesting that MZ09 represents a potentially promising therapeutic agent for IBD treatment.
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.
