Autophagy is well-known for delivering cargo materials to lysosomes for proteolytic digestion.Recently,autophagy has emerged as a key mechanism in unconventional protein secretion(UPS).This perspective introduces unco...Autophagy is well-known for delivering cargo materials to lysosomes for proteolytic digestion.Recently,autophagy has emerged as a key mechanism in unconventional protein secretion(UPS).This perspective introduces unconventional secretion pathways,focusing on secretory autophagy and its role in secreting protein aggregates associated with neurodegenerative disorders.We also explore additional neuronal functions of secretory autophagy beyond the release of protein aggregates.We propose autophagosomes as transport organelles that deliver cargo material directly from the endoplasmatic reticulum(ER)to the plasma membrane rather than solely to lysosomes.展开更多
Objectives:Ribosomal protein S6 kinase A2(RPS6KA2)has been identified as a potential prognostic biomarker in several cancers,including breast cancer,glioblastoma,and prostate cancer.However,its functional significance...Objectives:Ribosomal protein S6 kinase A2(RPS6KA2)has been identified as a potential prognostic biomarker in several cancers,including breast cancer,glioblastoma,and prostate cancer.However,its functional significance in ovarian cancer is not well characterized.This study was designed to explore the therapeutic relevance of modulating RPS6KA2 in the context of ovarian cancer,particularly in relation to cisplatin resistance.Methods:The expression levels of RPS6KA2 and key regulators involved in autophagy and ferroptosis were assessed using quantitative reverse transcription-PCR,immunofluorescence staining,immunohistochemistry,and western blotting.Prognostic associations were conducted using the Kaplan-Meier Plotter database.Autophagy flux assays and visualization of autophagosomes were performed to assess autophagy activity.Ferroptosis-related parameters,including intracellular iron content,glutathione(GSH)levels,reactive oxygen species(ROS)generation,and mitochondrial membrane potential,were measured to determine ferroptotic changes.In vivo experiments were carried out to determine the antitumor efficacy of RPS6KA2 modulation in combination with pathway-specific agents.Results:Using ovarian cancer cell lines and clinical tissue samples,we demonstrated that RPS6KA2 expression was significantly downregulated in cisplatin-resistant cells and tissues compared to their sensitive counterparts.Low RPS6KA2 expression correlated with unfavorable patient outcomes and enhanced chemoresistance.Mechanistically,RPS6KA2 inhibited autophagy by modulating the phosphatidylinositol 3-kinase-protein kinase B-mammalian target of rapamycin(PI3K-AKT-mTOR)signaling pathway,which in turn increased sensitivity to cisplatin.Additionally,RPS6KA2 facilitated ferroptosis,contributing to its tumor-suppressive function.miR-512-3p was identified as a negative regulator of RPS6KA2,driving cisplatin resistance through suppression of RPS6KA2 expression.In vivo validation confirmed that combining RPS6KA2 targeting with autophagy inhibitors or ferroptosis inducers significantly enhanced cisplatin sensitivity in ovarian cancer models.Conclusion:These results collectively indicate that targeting the miR-512-3p/RPS6KA2 regulatory axis may offer a novel and effective strategy for overcoming cisplatin resistance in ovarian cancer.展开更多
The endoplasmic reticulum,a key cellular organelle,regulates a wide variety of cellular activities.Endoplasmic reticulum autophagy,one of the quality control systems of the endoplasmic reticulum,plays a pivotal role i...The endoplasmic reticulum,a key cellular organelle,regulates a wide variety of cellular activities.Endoplasmic reticulum autophagy,one of the quality control systems of the endoplasmic reticulum,plays a pivotal role in maintaining endoplasmic reticulum homeostasis by controlling endoplasmic reticulum turnover,remodeling,and proteostasis.In this review,we briefly describe the endoplasmic reticulum quality control system,and subsequently focus on the role of endoplasmic reticulum autophagy,emphasizing the spatial and temporal mechanisms underlying the regulation of endoplasmic reticulum autophagy according to cellular requirements.We also summarize the evidence relating to how defective or abnormal endoplasmic reticulum autophagy contributes to the pathogenesis of neurodegenerative diseases.In summary,this review highlights the mechanisms associated with the regulation of endoplasmic reticulum autophagy and how they influence the pathophysiology of degenerative nerve disorders.This review would help researchers to understand the roles and regulatory mechanisms of endoplasmic reticulum-phagy in neurodegenerative disorders.展开更多
Nerve regeneration following traumatic peripheral nerve injuries and neuropathies is a complex process modulated by diverse factors and intricate molecular mechanisms.Past studies have focused on factors that stimulat...Nerve regeneration following traumatic peripheral nerve injuries and neuropathies is a complex process modulated by diverse factors and intricate molecular mechanisms.Past studies have focused on factors that stimulate axonal outgrowth and myelin regeneration.However,recent studies have highlighted the pivotal role of autophagy in peripheral nerve regeneration,particularly in the context of traumatic injuries.Consequently,autophagy-targeting modulation has emerged as a promising therapeutic approach to enhancing peripheral nerve regeneration.Our current understanding suggests that activating autophagy facilitates the rapid clearance of damaged axons and myelin sheaths,thereby enhancing neuronal survival and mitigating injury-induced oxidative stress and inflammation.These actions collectively contribute to creating a favorable microenvironment for structural and functional nerve regeneration.A range of autophagyinducing drugs and interventions have demonstrated beneficial effects in alleviating peripheral neuropathy and promoting nerve regeneration in preclinical models of traumatic peripheral nerve injuries.This review delves into the regulation of autophagy in cell types involved in peripheral nerve regeneration,summarizing the potential drugs and interventions that can be harnessed to promote this process.We hope that our review will offer novel insights and perspectives on the exploitation of autophagy pathways in the treatment of peripheral nerve injuries and neuropathies.展开更多
Ovarian cancer(OC),a common malignancy of the female reproductive system,has the highest mortality rate among gynecological cancers.A distinguishing feature of OC cells(OCCs)is their reduced autophagic flux compared w...Ovarian cancer(OC),a common malignancy of the female reproductive system,has the highest mortality rate among gynecological cancers.A distinguishing feature of OC cells(OCCs)is their reduced autophagic flux compared with normal cells.This phenomenon indicates that excessive autophagy activation or impaired autophagosome–lysosome fusion may lead to OCC death.This study investigated the anti-OC effects of dihydrotanshinone I(DHT),a tanshinone compound from Salvia miltiorrhiza.Proteomic analysis suggested that DHT suppressed OC growth via the autophagy–lysosome pathway,with sortilin 1(SORT1)identified as a critical target.In vitro,DHT promoted autophagosome formation mediated by microtubule-associated protein 1 light chain 3-II(LC3-II),while inhibiting autophagosome–lysosome fusion.The results of an orthotopic OC model corroborated these findings,showing that DHT induced autophagic cell death(ACD)and suppressed SORT1 expression in tumors.Further RNA interference experiments confirmed that SORT1 depletion caused autophagosomes to accumulate in OCCs.Notably,we found that SORT1 interacted with autophagy-related gene(ATG)-encoded proteins ATG5 and ATG16L1,and that depleting SORT1 increased the levels of these proteins.Co-immunoprecipitation,ubiquitination,and cellular thermal shift assay analyses revealed that DHT directly targeted and promoted ubiquitin-dependent degradation of SORT1.By degrading SORT1,ATG5 and ATG16L1 were released,which enhanced autophagosome formation and disrupted the autophagic flux.These findings identified DHT as a novel autophagosome inducer that induced ACD by targeting SORT1,making it a promising therapeutic candidate for OC.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
Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cereb...Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cerebral ischemia,but the relationship between ER stress and autophagy remains unclear.In this study,we established experimental models using oxygen-glucose deprivation/reoxygenation in PC12 cells and primary neurons to simulate cerebral ischemia/reperfusion injury.We found that prolongation of oxygen-glucose deprivation activated the ER stress pathway protein kinase-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2 subunit alpha(e IF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP),increased neuronal apoptosis,and induced autophagy.Furthermore,inhibition of ER stress using inhibitors or by si RNA knockdown of the PERK gene significantly attenuated excessive autophagy and neuronal apoptosis,indicating an interaction between autophagy and ER stress and suggesting PERK as an essential target for regulating autophagy.Blocking autophagy with chloroquine exacerbated ER stress-induced apoptosis,indicating that normal levels of autophagy play a protective role in neuronal injury following cerebral ischemia/reperfusion injury.Findings from this study indicate that cerebral ischemia/reperfusion injury can trigger neuronal ER stress and promote autophagy,and suggest that PERK is a possible target for inhibiting excessive autophagy in cerebral ischemia/reperfusion injury.展开更多
Salsolinol(1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline,Sal)is a catechol isoquinoline that causes neurotoxicity and shares structural similarity with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,an environme...Salsolinol(1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline,Sal)is a catechol isoquinoline that causes neurotoxicity and shares structural similarity with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,an environmental toxin that causes Parkinson's disease.However,the mechanism by which Sal mediates dopaminergic neuronal death remains unclear.In this study,we found that Sal significantly enhanced the global level of N~6-methyladenosine(m~6A)RNA methylation in PC12 cells,mainly by inducing the downregulation of the expression of m~6A demethylases fat mass and obesity-associated protein(FTO)and alk B homolog 5(ALKBH5).RNA sequencing analysis showed that Sal downregulated the Hippo signaling pathway.The m~6A reader YTH domain-containing family protein 2(YTHDF2)promoted the degradation of m~6A-containing Yes-associated protein 1(YAP1)mRNA,which is a downstream key effector in the Hippo signaling pathway.Additionally,downregulation of YAP1 promoted autophagy,indicating that the mutual regulation between YAP1 and autophagy can lead to neurotoxicity.These findings reveal the role of Sal on m~6A RNA methylation and suggest that Sal may act as an RNA methylation inducer mediating dopaminergic neuronal death through YAP1 and autophagy.Our results provide greater insights into the neurotoxic effects of catechol isoquinolines compared with other studies and may be a reference for assessing the involvement of RNA methylation in the pathogenesis of Parkinson's disease.展开更多
Parkinson’s disease is a common neurodegenerative disease with movement disorders associated with the intracytoplasmic deposition of aggregate proteins such asα-synuclein in neurons.As one of the major intracellular...Parkinson’s disease is a common neurodegenerative disease with movement disorders associated with the intracytoplasmic deposition of aggregate proteins such asα-synuclein in neurons.As one of the major intracellular degradation pathways,the autophagy-lysosome pathway plays an important role in eliminating these proteins.Accumulating evidence has shown that upregulation of the autophagy-lysosome pathway may contribute to the clearance ofα-synuclein aggregates and protect against degeneration of dopaminergic neurons in Parkinson’s disease.Moreover,multiple genes associated with the pathogenesis of Parkinson’s disease are intimately linked to alterations in the autophagy-lysosome pathway.Thus,this pathway appears to be a promising therapeutic target for treatment of Parkinson’s disease.In this review,we briefly introduce the machinery of autophagy.Then,we provide a description of the effects of Parkinson’s disease–related genes on the autophagy-lysosome pathway.Finally,we highlight the potential chemical and genetic therapeutic strategies targeting the autophagy–lysosome pathway and their applications in Parkinson’s disease.展开更多
BACKGROUND Runchangningshen paste(RCNSP)is a paste made of four medicinal and edible homologous Chinese medicine mixed with honey.It is known for its ability to nourish yin and blood as well as to loosen the bowel to ...BACKGROUND Runchangningshen paste(RCNSP)is a paste made of four medicinal and edible homologous Chinese medicine mixed with honey.It is known for its ability to nourish yin and blood as well as to loosen the bowel to relieve constipation.The pathophysiology of functional constipation(FC)is associated with a reduction in mucin-2(MUC2)secretion and microbial dysbiosis.AIM To investigate the underlying mechanism of RCNSP against FC through MUC2 and the gut mucosal microbiota.METHODS Ultra-performance liquid chromatography tandem mass spectrometry characterized RCNSP composition to elucidate the material basis of action.FC model was induced via loperamide gavage(16 mg/kg)twice daily for 7 days.Applying defecation function and gastrointestinal motility to assess constipation severity.Hematoxylin and eosin and Alcian blue-periodic acid-schiff staining analyzed colonic mucosal morphology.Transmission electron microscope was used to observe the ultrastructure of goblet cells(GCs).Immunofluorescence colocalization,quantitative PCR,and western blot assessed the impact of RCNSP on gene and protein expression within the NLRP6/autophagy pathway.16S rDNA was employed to sequence the gut mucosal microbiota.RESULTS RCNSP contained 12 components with potential laxative effects.It enhanced defecation function,accelerated gastrointestinal motility,and maintained colonic mucosal integrity.RCNSP treatment significantly increased GC abundance and MUC2 production while preserving GC ultrastructure.At the molecular level,RCNSP enhanced the colocalized expression of key regulatory proteins and modulated mRNA and protein expressions in the NLRP6/autophagy pathway.Through 16S rDNA sequencing analysis,RCNSP significantly altered the mucosal microbiota composition.Specifically,it increased beneficial bacterial strains while reducing harmful ones.Simultaneously,RCNSP reduced butyrate-producing bacteria like Proteobacteria,Enterobacteriaceae,Blautia,and Eubacterium and decreased hydrogen sulfide-producing species,such as Prevotellaceae.It also reduced bile acidinhibiting species,such as g_Eubacter_coprostanoligenes_group and Erysipelotrichaceae while increasing bile acidproducing species,such as Colidextribacter.CONCLUSION Our findings suggested that RCNSP ameliorated constipation through a dual mechanism:It stimulated colonic MUC2 secretion by activating NLRP6 inflammasome-mediated autophagy and modulated the composition of the mucosal microbiota.展开更多
The intestinal tract,a complex organ responsible for nutrient absorption and digestion,relies heavily on a balanced gut microbiome to maintain its integrity.Disruptions to this delicate microbial ecosystem can lead to...The intestinal tract,a complex organ responsible for nutrient absorption and digestion,relies heavily on a balanced gut microbiome to maintain its integrity.Disruptions to this delicate microbial ecosystem can lead to intestinal inflammation,a hallmark of inflammatory bowel disease(IBD).While the role of the gut microbiome in IBD is increasingly recognized,the underlying mechanisms,particularly those involving endoplasmic reticulum(ER)stress,autophagy,and cell death,remain incompletely understood.ER stress,a cellular response to various stressors,can trigger inflammation and cell death.Autophagy,a cellular degradation process,can either alleviate or exacerbate ER stress-induced inflammation,depending on the specific context.The gut microbiome can influence both ER stress and autophagy pathways,further complicating the interplay between these processes.This review delves into the intricate relationship between ER stress,autophagy,and the gut microbiome in the context of intestinal inflammation.By exploring the molecular mechanisms underlying these interactions,we aim to provide a comprehensive theoretical framework for developing novel therapeutic strategies for IBD.A deeper understanding of the ER stress-autophagy axis,the gut microbial-ER stress axis,and the gut microbial-autophagy axis may pave the way for targeted interventions to restore intestinal health and mitigate the impact of IBD.展开更多
Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,...Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,where it may play a role in either promoting or inhibiting tumor growth.This study aimed to investigate the expression level,biological functions,and molecular mechanisms of EMP2 in liver cancer.Methods:we analyzed the mRNA expression levels of EMPs family genes in hepatocellular carcinoma(HCC)tissues and normal liver tissues based on the TCGA database and immunohistochemical analysis of tissue microarrays.Subsequently,we constructed HCC cell lines with either knockdown or overexpression of EMP2 to examine the biological functions and molecular mechanisms of EMP2 in tumorigenesis in vivo and in vitro.Results:Bioinformatic and immunohistochemical analysis of tissue microarrays have confirmed the significant upregulation of EMP2 in HCC tissues.In vitro and in vivo studies have shown that downregulation of EMP2 results in a moderate reduction in the proliferation and invasive capacity of HCC cells.Conversely,overexpression of EMP2 enhances the invasive capacity of HCC cells and induces autophagy.Initial investigations into the molecular mechanisms underlying EMP2-mediated enhancement of HCC cell invasion have revealed the dual regulation of EMP2-induced autophagy and the integrin pathway,which synergistically influence the invasive and metastatic potential of HCC cells.Conclusion:EMP2 holds promise as a diagnostic marker for HCC metastasis and a potential target for targeted therapy.展开更多
Cells undergo metabolic reprogramming to adapt to changes in nutrient availability, cellular activity, and transitions in cell states. The balance between glycolysis and mitochondrial respiration is crucial for energy...Cells undergo metabolic reprogramming to adapt to changes in nutrient availability, cellular activity, and transitions in cell states. The balance between glycolysis and mitochondrial respiration is crucial for energy production, and metabolic reprogramming stipulates a shift in such balance to optimize both bioenergetic efficiency and anabolic requirements. Failure in switching bioenergetic dependence can lead to maladaptation and pathogenesis. While cellular degradation is known to recycle precursor molecules for anabolism, its potential role in regulating energy production remains less explored. The bioenergetic switch between glycolysis and mitochondrial respiration involves transcription factors and organelle homeostasis, which are both regulated by the cellular degradation pathways. A growing body of studies has demonstrated that both stem cells and differentiated cells exhibit bioenergetic switch upon perturbations of autophagic activity or endolysosomal processes. Here, we highlighted the current understanding of the interplay between degradation processes, specifically autophagy and endolysosomes, transcription factors, endolysosomal signaling, and mitochondrial homeostasis in shaping cellular bioenergetics. This review aims to summarize the relationship between degradation processes and bioenergetics, providing a foundation for future research to unveil deeper mechanistic insights into bioenergetic regulation.展开更多
Objective:Oxaliplatin(OXA)and 5-fluorouracil(5-FU)are 2 commonly used chemotherapeutic agents for colorectal cancer(CRC).MicroRNAs(miRNAs,miRs)play crucial roles in the development of chemoresistance in various cancer...Objective:Oxaliplatin(OXA)and 5-fluorouracil(5-FU)are 2 commonly used chemotherapeutic agents for colorectal cancer(CRC).MicroRNAs(miRNAs,miRs)play crucial roles in the development of chemoresistance in various cancers.However,the role and mechanism of miR-224-5p in regulating CRC chemoresistance remain unclear.This study aims to investigate the function of miR-224-5p in chemoresistant CRC cells and the underlying mechanisms.Methods:CRC datasets GSE28702 and GSE69657 were downloaded from the Gene Expression Omnibus(GEO)database.Differentially expressed miRNAs between drug sensitive and resistant groups(OXA or 5-FU)were analyzed,and miR-224-5p was identified as the target miRNA.Chemoresistant cell lines HCT15-OXR,HCT15-5-FU,SW480-OXR,and SW480-5-FU were established.Transient transfections were performed using miR-224-5p mimics,inhibitors,and their respective negative controls(control mimic,control inhibitor)in these cell lines.Cells were treated with different concentrations of OXA or 5-FU post-transfection,and the half-maximal inhibitory concentration(IC_(50))was determined using the cell counting kit-8(CCK-8)assay.Cell proliferation was assessed by CCK-8 and colony formation assays.The expression levels of miR-224-5p,LC3,and P62 were measured by real-time polymerase chain reaction(real-time PCR)and/or Western blotting.Autophagic flux was assessed using a tandem fluorescent-tagged LC3 reporter assay.TargetScan 8.0,miRTarBase,miRPathDB,and HADb were used to predict B-cell lymphoma-2(Bcl-2)as a potential miR-244-5p target,which was further validated by dual luciferase reporter assays.Results:Chemoresistant CRC cells exhibited down-regulated miR-224-5p expression,whereas up-regulation of miR-224-5p enhanced chemotherapy sensitivity.Exposure to OXA or 5-FU significantly increased autophagic activity in chemoresistant CRC cells,which was reversed by miR-224-5p overexpression.Dual-luciferase assays verified Bcl-2 as a direct target of miR-224-5p.Conclusion:MiR-224-5p regulates chemoresistance in CRC by modulating autophagy through direct targeting of Bcl-2.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
Oligodendrocyte lineage cells, including oligodendrocyte precursor cells (OPCs) and oligodendrocytes (OLs), are essential in establishing and maintaining brain circuits. Autophagy is a conserved process that keeps the...Oligodendrocyte lineage cells, including oligodendrocyte precursor cells (OPCs) and oligodendrocytes (OLs), are essential in establishing and maintaining brain circuits. Autophagy is a conserved process that keeps the quality of organelles and proteostasis. The role of autophagy in oligodendrocyte lineage cells remains unclear. The present study shows that autophagy is required to maintain the number of OPCs/OLs and myelin integrity during brain aging. Inactivation of autophagy in oligodendrocyte lineage cells increases the number of OPCs/OLs in the developing brain while exaggerating the loss of OPCs/OLs with brain aging. Inactivation of autophagy in oligodendrocyte lineage cells impairs the turnover of myelin basic protein (MBP). It causes MBP to accumulate in the cytoplasm as multimeric aggregates and fails to be incorporated into integral myelin, which is associated with attenuated endocytic recycling. Inactivation of autophagy in oligodendrocyte lineage cells impairs myelin integrity and causes demyelination. Thus, this study shows autophagy is required to maintain myelin quality during aging by controlling the turnover of myelin components.展开更多
Objective Electronic cigarettes(ECs)differ from traditional tobacco smoke but may contribute to cardiopulmonary remodeling.Pulmonary hypertension(PH),characterized by pulmonary artery and right ventricle remodeling,po...Objective Electronic cigarettes(ECs)differ from traditional tobacco smoke but may contribute to cardiopulmonary remodeling.Pulmonary hypertension(PH),characterized by pulmonary artery and right ventricle remodeling,poses a significant risk of mortality in infants,children,and adolescents.However,the impact of maternal EC exposure on PH development in offspring remains unclear.To address this,we established a PH rat model with maternal EC exposure.Methods Maternal EC exposure was initiated on gestation day 12 via electronic nicotine delivery systems.Offspring were administered monocrotaline(MCT)at 6 weeks of age(6-wo)to induce PH.Mechanistic experiments were conducted at 10-week-old(10-wo).Protein expression of NADPH oxidases,DNA methyltransferases,and autophagy-related markers was analyzed by Western blot.Morphological changes and the severity of PH were evaluated via hematoxylin and eosin(HE)staining and echocardiography,respectively.Furthermore,the involvement of the oxidative stress/DNA methylation/autophagy axis in response to maternal EC exposure was confirmed through a combination of ELISA,Western blot,HE staining,and echocardiography.Additionally,ATG5 mRNA expression was measured by qRT-PCR.Results Compared with control conditions,maternal EC exposure significantly worsened MCT-induced PH in male offspring.This was associated with increased oxidative stress,DNA hypomethylation,and anomalous autophagy in the offspring.In vivo treatment with chloroquine inhibited autophagy and ameliorated PH development in offspring exposed to maternal EC.Furthermore,N-acetylcysteine(NAC),an antioxidant,attenuated maternal EC exposure-induced oxidative stress,DNA hypomethylation,and excessive autophagy,thereby improving PH.DNA hypermethylation also reversed PH development,accompanied by reduced oxidative stress and suppressed autophagy.ATG5,a key regulator of autophagy,was identified as a potential therapeutic target,as its repression mitigated PH in maternal EC-exposed offspring.Conclusion Maternal EC exposure induces oxidative stress and DNA hypomethylation in offspring,leading to anomalous autophagy and exacerbation of PH development.Targeting ATG5-mediated autophagy may represent a novel therapeutic approach for improving PH outcomes in offspring exposed to maternal EC.Graphical Abstract Pregnant rats were exposed to either EC vapor or standard air from gestation day 12 until 2 days before delivery,with all offspring undergoing PH induction at 6-wo.Offspring exposed to maternal EC presented increased oxidative stress,which in turn affected DNA methylation patterns.The decreased DNA methylation in male offspring led to the activation of autophagy,exacerbating the development of PH.Treatment with ATG5 siRNA inhibited autophagy and alleviated heightened PH in male offspring with maternal EC exposure.展开更多
Mycobacterium tuberculosis(Mtb)employs multiple mechanisms,such as phagocytosis and autophagy,to evade innate immune clearance and establish infection.In the present study,we identified the ESX-1 secretion-associated ...Mycobacterium tuberculosis(Mtb)employs multiple mechanisms,such as phagocytosis and autophagy,to evade innate immune clearance and establish infection.In the present study,we identified the ESX-1 secretion-associated protein EspL,which promotes Mtb survival by inhibiting phagosome maturation and autophagy initiation.EspL knockout decreased Mtb intracellular survival,while EspL overexpression increased bacterial survival by interfering with phagocytosis and autophagy.EspL interacts with ULK1 and promotes its phosphorylation at Ser^(757),leading to the inhibition of autophagy initiation.Additionally,overexpression of EspL reduced antigen presentation and T-cell responses both in vitro and in vivo.Our findings revealed that EspL interferes with autophagy and antigen presenta-tion by suppressing ULK1 activation.These insights provide a novel understanding of Mtb pathogenicity.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
Autophagy is a universal cellular process in eukaryotes that plays a critical role in plant growth and stress response.However,the role of autophagy in fruit ripening is largely unknown.Here,we demonstrated that most ...Autophagy is a universal cellular process in eukaryotes that plays a critical role in plant growth and stress response.However,the role of autophagy in fruit ripening is largely unknown.Here,we demonstrated that most autophagy-related genes(ATGs)were up-regulated during tomato(Solanum lycopersicum L.)fruit ripening.By using mutants of different autophagy pathway genes(ATG6,ATG10,ATG18a),we revealed that the deficiency of autophagy delayed the ripening of fruit.Compared with wild-type(WT),the production of ethylene was significantly reduced and the accumulation of lycopene was delayed in atg mutants during fruit ripening.We also observed the contents of glucose and fructose were both significantly decreased in atg mutants compared with WT,while the content of organic acids showed the opposite trend.Additionally,the negative regulator of ethylene production,APETALA2a(AP2a),interacted with ATG8 through a specific ATG8-interacting motif(AIM)and could be degraded through the autophagy pathway.These results demonstrate that autophagy plays a critical role in fruit ripening by regulating ethylene production and the accumulation of pigments,sugars and organic acids in tomato.展开更多
基金supported by the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)grant LU 2347/3-1(to PL).
文摘Autophagy is well-known for delivering cargo materials to lysosomes for proteolytic digestion.Recently,autophagy has emerged as a key mechanism in unconventional protein secretion(UPS).This perspective introduces unconventional secretion pathways,focusing on secretory autophagy and its role in secreting protein aggregates associated with neurodegenerative disorders.We also explore additional neuronal functions of secretory autophagy beyond the release of protein aggregates.We propose autophagosomes as transport organelles that deliver cargo material directly from the endoplasmatic reticulum(ER)to the plasma membrane rather than solely to lysosomes.
基金supported by the Academic Leader Training Programof Pudong New Area Health System in Shanghai(Grant No.PWRd2021-13)Shanghai Municipal Health Commission(Grant No.202340094).
文摘Objectives:Ribosomal protein S6 kinase A2(RPS6KA2)has been identified as a potential prognostic biomarker in several cancers,including breast cancer,glioblastoma,and prostate cancer.However,its functional significance in ovarian cancer is not well characterized.This study was designed to explore the therapeutic relevance of modulating RPS6KA2 in the context of ovarian cancer,particularly in relation to cisplatin resistance.Methods:The expression levels of RPS6KA2 and key regulators involved in autophagy and ferroptosis were assessed using quantitative reverse transcription-PCR,immunofluorescence staining,immunohistochemistry,and western blotting.Prognostic associations were conducted using the Kaplan-Meier Plotter database.Autophagy flux assays and visualization of autophagosomes were performed to assess autophagy activity.Ferroptosis-related parameters,including intracellular iron content,glutathione(GSH)levels,reactive oxygen species(ROS)generation,and mitochondrial membrane potential,were measured to determine ferroptotic changes.In vivo experiments were carried out to determine the antitumor efficacy of RPS6KA2 modulation in combination with pathway-specific agents.Results:Using ovarian cancer cell lines and clinical tissue samples,we demonstrated that RPS6KA2 expression was significantly downregulated in cisplatin-resistant cells and tissues compared to their sensitive counterparts.Low RPS6KA2 expression correlated with unfavorable patient outcomes and enhanced chemoresistance.Mechanistically,RPS6KA2 inhibited autophagy by modulating the phosphatidylinositol 3-kinase-protein kinase B-mammalian target of rapamycin(PI3K-AKT-mTOR)signaling pathway,which in turn increased sensitivity to cisplatin.Additionally,RPS6KA2 facilitated ferroptosis,contributing to its tumor-suppressive function.miR-512-3p was identified as a negative regulator of RPS6KA2,driving cisplatin resistance through suppression of RPS6KA2 expression.In vivo validation confirmed that combining RPS6KA2 targeting with autophagy inhibitors or ferroptosis inducers significantly enhanced cisplatin sensitivity in ovarian cancer models.Conclusion:These results collectively indicate that targeting the miR-512-3p/RPS6KA2 regulatory axis may offer a novel and effective strategy for overcoming cisplatin resistance in ovarian cancer.
基金supported by the National Natural Science Foundation of China,Nos.92049120 and 81870897STI2030-Major Projects,No.2021ZD0204001+6 种基金Guangdong Key Project for Development of New Tools for the Diagnosis and Treatment of Autism,No.2018B030335001the Natural Science Foundation of Jiangsu Province,No.BK20181436the National Major Scientific and Technological Special Project for Significant New Drug Development,No.2019ZX09301102the Discipline Construction Program of the Second Affiliated Hospital of Soochow University,No.XKTJ-TD202003Sino-German Cooperation Mobility Programme,No.M-0679the Science and Technology Project of Suzhou,No.SKY2022161Research Project of Neurological Diseases of the Second Affiliated Hospital of Soochow University Medical Center,No.ND2023A01(all to QHM)。
文摘The endoplasmic reticulum,a key cellular organelle,regulates a wide variety of cellular activities.Endoplasmic reticulum autophagy,one of the quality control systems of the endoplasmic reticulum,plays a pivotal role in maintaining endoplasmic reticulum homeostasis by controlling endoplasmic reticulum turnover,remodeling,and proteostasis.In this review,we briefly describe the endoplasmic reticulum quality control system,and subsequently focus on the role of endoplasmic reticulum autophagy,emphasizing the spatial and temporal mechanisms underlying the regulation of endoplasmic reticulum autophagy according to cellular requirements.We also summarize the evidence relating to how defective or abnormal endoplasmic reticulum autophagy contributes to the pathogenesis of neurodegenerative diseases.In summary,this review highlights the mechanisms associated with the regulation of endoplasmic reticulum autophagy and how they influence the pathophysiology of degenerative nerve disorders.This review would help researchers to understand the roles and regulatory mechanisms of endoplasmic reticulum-phagy in neurodegenerative disorders.
基金supported by the National Natural Science Foundation of China,Nos.82271411(to RG),51803072(to WLiu)grants from the Department of Finance of Jilin Province,Nos.2022SCZ25(to RG),2022SCZ10(to WLiu),2021SCZ07(to RG)+2 种基金Jilin Provincial Science and Technology Program,No.YDZJ202201ZYTS038(to WLiu)The Youth Support Programmed Project of China-Japan Union Hospital of Jilin University,No.2022qnpy11(to WLuo)The Project of China-Japan Union Hospital of Jilin University,No.XHQMX20233(to RG)。
文摘Nerve regeneration following traumatic peripheral nerve injuries and neuropathies is a complex process modulated by diverse factors and intricate molecular mechanisms.Past studies have focused on factors that stimulate axonal outgrowth and myelin regeneration.However,recent studies have highlighted the pivotal role of autophagy in peripheral nerve regeneration,particularly in the context of traumatic injuries.Consequently,autophagy-targeting modulation has emerged as a promising therapeutic approach to enhancing peripheral nerve regeneration.Our current understanding suggests that activating autophagy facilitates the rapid clearance of damaged axons and myelin sheaths,thereby enhancing neuronal survival and mitigating injury-induced oxidative stress and inflammation.These actions collectively contribute to creating a favorable microenvironment for structural and functional nerve regeneration.A range of autophagyinducing drugs and interventions have demonstrated beneficial effects in alleviating peripheral neuropathy and promoting nerve regeneration in preclinical models of traumatic peripheral nerve injuries.This review delves into the regulation of autophagy in cell types involved in peripheral nerve regeneration,summarizing the potential drugs and interventions that can be harnessed to promote this process.We hope that our review will offer novel insights and perspectives on the exploitation of autophagy pathways in the treatment of peripheral nerve injuries and neuropathies.
基金supported by the National Key Research and Development Program of China(2023YFC3503900)the National Natural Science Foundation of China(82305001)+3 种基金the Zhejiang Provincial Natural Science Foundation of China(LQ24H280011)the Science Research Fund of Administration of Traditional Chinese Medicine of Zhejiang Province(2023ZR014)the National Young Qihuang Scholars Training Programthe Research Project of Zhejiang Chinese Medical University(2022RCZXZK18,2023JKZKTS17)。
文摘Ovarian cancer(OC),a common malignancy of the female reproductive system,has the highest mortality rate among gynecological cancers.A distinguishing feature of OC cells(OCCs)is their reduced autophagic flux compared with normal cells.This phenomenon indicates that excessive autophagy activation or impaired autophagosome–lysosome fusion may lead to OCC death.This study investigated the anti-OC effects of dihydrotanshinone I(DHT),a tanshinone compound from Salvia miltiorrhiza.Proteomic analysis suggested that DHT suppressed OC growth via the autophagy–lysosome pathway,with sortilin 1(SORT1)identified as a critical target.In vitro,DHT promoted autophagosome formation mediated by microtubule-associated protein 1 light chain 3-II(LC3-II),while inhibiting autophagosome–lysosome fusion.The results of an orthotopic OC model corroborated these findings,showing that DHT induced autophagic cell death(ACD)and suppressed SORT1 expression in tumors.Further RNA interference experiments confirmed that SORT1 depletion caused autophagosomes to accumulate in OCCs.Notably,we found that SORT1 interacted with autophagy-related gene(ATG)-encoded proteins ATG5 and ATG16L1,and that depleting SORT1 increased the levels of these proteins.Co-immunoprecipitation,ubiquitination,and cellular thermal shift assay analyses revealed that DHT directly targeted and promoted ubiquitin-dependent degradation of SORT1.By degrading SORT1,ATG5 and ATG16L1 were released,which enhanced autophagosome formation and disrupted the autophagic flux.These findings identified DHT as a novel autophagosome inducer that induced ACD by targeting SORT1,making it a promising therapeutic candidate for OC.
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 National Natural Science Foundation of China,Nos.82260245(to YX),81660207(to YX),81960253(to YL),82160268(to YL),U1812403(to ZG)Science and Technology Projects of Guizhou Province,Nos.[2019]1440(to YX),[2020]1Z067(to WH)+1 种基金Cultivation Foundation of Guizhou Medical University,No.[20NSP069](to YX)Excellent Young Talents Plan of Guizhou Medical University,No.(2022)101(to WH)。
文摘Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cerebral ischemia,but the relationship between ER stress and autophagy remains unclear.In this study,we established experimental models using oxygen-glucose deprivation/reoxygenation in PC12 cells and primary neurons to simulate cerebral ischemia/reperfusion injury.We found that prolongation of oxygen-glucose deprivation activated the ER stress pathway protein kinase-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2 subunit alpha(e IF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP),increased neuronal apoptosis,and induced autophagy.Furthermore,inhibition of ER stress using inhibitors or by si RNA knockdown of the PERK gene significantly attenuated excessive autophagy and neuronal apoptosis,indicating an interaction between autophagy and ER stress and suggesting PERK as an essential target for regulating autophagy.Blocking autophagy with chloroquine exacerbated ER stress-induced apoptosis,indicating that normal levels of autophagy play a protective role in neuronal injury following cerebral ischemia/reperfusion injury.Findings from this study indicate that cerebral ischemia/reperfusion injury can trigger neuronal ER stress and promote autophagy,and suggest that PERK is a possible target for inhibiting excessive autophagy in cerebral ischemia/reperfusion injury.
基金supported by the National Natural Science Foundation of China,Nos.82271283(to XC),91854115(to JW),31970044(to JW)the Natural Science Foundation of Beijing,No.7202001(to XC)the Scientific Research Project of Beijing Educational Committee,No.KM202010005022(to XC)。
文摘Salsolinol(1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline,Sal)is a catechol isoquinoline that causes neurotoxicity and shares structural similarity with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,an environmental toxin that causes Parkinson's disease.However,the mechanism by which Sal mediates dopaminergic neuronal death remains unclear.In this study,we found that Sal significantly enhanced the global level of N~6-methyladenosine(m~6A)RNA methylation in PC12 cells,mainly by inducing the downregulation of the expression of m~6A demethylases fat mass and obesity-associated protein(FTO)and alk B homolog 5(ALKBH5).RNA sequencing analysis showed that Sal downregulated the Hippo signaling pathway.The m~6A reader YTH domain-containing family protein 2(YTHDF2)promoted the degradation of m~6A-containing Yes-associated protein 1(YAP1)mRNA,which is a downstream key effector in the Hippo signaling pathway.Additionally,downregulation of YAP1 promoted autophagy,indicating that the mutual regulation between YAP1 and autophagy can lead to neurotoxicity.These findings reveal the role of Sal on m~6A RNA methylation and suggest that Sal may act as an RNA methylation inducer mediating dopaminergic neuronal death through YAP1 and autophagy.Our results provide greater insights into the neurotoxic effects of catechol isoquinolines compared with other studies and may be a reference for assessing the involvement of RNA methylation in the pathogenesis of Parkinson's disease.
基金supported by the National Natural Science Foundation of China,No.82101340(to FJ).
文摘Parkinson’s disease is a common neurodegenerative disease with movement disorders associated with the intracytoplasmic deposition of aggregate proteins such asα-synuclein in neurons.As one of the major intracellular degradation pathways,the autophagy-lysosome pathway plays an important role in eliminating these proteins.Accumulating evidence has shown that upregulation of the autophagy-lysosome pathway may contribute to the clearance ofα-synuclein aggregates and protect against degeneration of dopaminergic neurons in Parkinson’s disease.Moreover,multiple genes associated with the pathogenesis of Parkinson’s disease are intimately linked to alterations in the autophagy-lysosome pathway.Thus,this pathway appears to be a promising therapeutic target for treatment of Parkinson’s disease.In this review,we briefly introduce the machinery of autophagy.Then,we provide a description of the effects of Parkinson’s disease–related genes on the autophagy-lysosome pathway.Finally,we highlight the potential chemical and genetic therapeutic strategies targeting the autophagy–lysosome pathway and their applications in Parkinson’s disease.
基金Supported by National Natural Science Foundation of China,No.82174309and Shanghai Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine Siming Fund Special Project for Scientific Research,No.SGKJ-202304.
文摘BACKGROUND Runchangningshen paste(RCNSP)is a paste made of four medicinal and edible homologous Chinese medicine mixed with honey.It is known for its ability to nourish yin and blood as well as to loosen the bowel to relieve constipation.The pathophysiology of functional constipation(FC)is associated with a reduction in mucin-2(MUC2)secretion and microbial dysbiosis.AIM To investigate the underlying mechanism of RCNSP against FC through MUC2 and the gut mucosal microbiota.METHODS Ultra-performance liquid chromatography tandem mass spectrometry characterized RCNSP composition to elucidate the material basis of action.FC model was induced via loperamide gavage(16 mg/kg)twice daily for 7 days.Applying defecation function and gastrointestinal motility to assess constipation severity.Hematoxylin and eosin and Alcian blue-periodic acid-schiff staining analyzed colonic mucosal morphology.Transmission electron microscope was used to observe the ultrastructure of goblet cells(GCs).Immunofluorescence colocalization,quantitative PCR,and western blot assessed the impact of RCNSP on gene and protein expression within the NLRP6/autophagy pathway.16S rDNA was employed to sequence the gut mucosal microbiota.RESULTS RCNSP contained 12 components with potential laxative effects.It enhanced defecation function,accelerated gastrointestinal motility,and maintained colonic mucosal integrity.RCNSP treatment significantly increased GC abundance and MUC2 production while preserving GC ultrastructure.At the molecular level,RCNSP enhanced the colocalized expression of key regulatory proteins and modulated mRNA and protein expressions in the NLRP6/autophagy pathway.Through 16S rDNA sequencing analysis,RCNSP significantly altered the mucosal microbiota composition.Specifically,it increased beneficial bacterial strains while reducing harmful ones.Simultaneously,RCNSP reduced butyrate-producing bacteria like Proteobacteria,Enterobacteriaceae,Blautia,and Eubacterium and decreased hydrogen sulfide-producing species,such as Prevotellaceae.It also reduced bile acidinhibiting species,such as g_Eubacter_coprostanoligenes_group and Erysipelotrichaceae while increasing bile acidproducing species,such as Colidextribacter.CONCLUSION Our findings suggested that RCNSP ameliorated constipation through a dual mechanism:It stimulated colonic MUC2 secretion by activating NLRP6 inflammasome-mediated autophagy and modulated the composition of the mucosal microbiota.
基金supported by the fund for the Project of the National Key Research and Development Program of China(2024YFD1300203)Project support was provided by the Fund opened from Key Laboratory of Fujian Universities Preventive Veterinary Medicine and Biotechnology,Longyan University(grant No.2021KF01)the Cyanine Project of Yangzhou University(2020)。
文摘The intestinal tract,a complex organ responsible for nutrient absorption and digestion,relies heavily on a balanced gut microbiome to maintain its integrity.Disruptions to this delicate microbial ecosystem can lead to intestinal inflammation,a hallmark of inflammatory bowel disease(IBD).While the role of the gut microbiome in IBD is increasingly recognized,the underlying mechanisms,particularly those involving endoplasmic reticulum(ER)stress,autophagy,and cell death,remain incompletely understood.ER stress,a cellular response to various stressors,can trigger inflammation and cell death.Autophagy,a cellular degradation process,can either alleviate or exacerbate ER stress-induced inflammation,depending on the specific context.The gut microbiome can influence both ER stress and autophagy pathways,further complicating the interplay between these processes.This review delves into the intricate relationship between ER stress,autophagy,and the gut microbiome in the context of intestinal inflammation.By exploring the molecular mechanisms underlying these interactions,we aim to provide a comprehensive theoretical framework for developing novel therapeutic strategies for IBD.A deeper understanding of the ER stress-autophagy axis,the gut microbial-ER stress axis,and the gut microbial-autophagy axis may pave the way for targeted interventions to restore intestinal health and mitigate the impact of IBD.
基金supported by the Fundamental Research Funds for the National Natural Science Foundation of China(Nos.22177084 and 82104373)the Fundamental Research Funds of Science&Technology Department of Sichuan Province(No.2022YFQ0054).
文摘Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,where it may play a role in either promoting or inhibiting tumor growth.This study aimed to investigate the expression level,biological functions,and molecular mechanisms of EMP2 in liver cancer.Methods:we analyzed the mRNA expression levels of EMPs family genes in hepatocellular carcinoma(HCC)tissues and normal liver tissues based on the TCGA database and immunohistochemical analysis of tissue microarrays.Subsequently,we constructed HCC cell lines with either knockdown or overexpression of EMP2 to examine the biological functions and molecular mechanisms of EMP2 in tumorigenesis in vivo and in vitro.Results:Bioinformatic and immunohistochemical analysis of tissue microarrays have confirmed the significant upregulation of EMP2 in HCC tissues.In vitro and in vivo studies have shown that downregulation of EMP2 results in a moderate reduction in the proliferation and invasive capacity of HCC cells.Conversely,overexpression of EMP2 enhances the invasive capacity of HCC cells and induces autophagy.Initial investigations into the molecular mechanisms underlying EMP2-mediated enhancement of HCC cell invasion have revealed the dual regulation of EMP2-induced autophagy and the integrin pathway,which synergistically influence the invasive and metastatic potential of HCC cells.Conclusion:EMP2 holds promise as a diagnostic marker for HCC metastasis and a potential target for targeted therapy.
文摘Cells undergo metabolic reprogramming to adapt to changes in nutrient availability, cellular activity, and transitions in cell states. The balance between glycolysis and mitochondrial respiration is crucial for energy production, and metabolic reprogramming stipulates a shift in such balance to optimize both bioenergetic efficiency and anabolic requirements. Failure in switching bioenergetic dependence can lead to maladaptation and pathogenesis. While cellular degradation is known to recycle precursor molecules for anabolism, its potential role in regulating energy production remains less explored. The bioenergetic switch between glycolysis and mitochondrial respiration involves transcription factors and organelle homeostasis, which are both regulated by the cellular degradation pathways. A growing body of studies has demonstrated that both stem cells and differentiated cells exhibit bioenergetic switch upon perturbations of autophagic activity or endolysosomal processes. Here, we highlighted the current understanding of the interplay between degradation processes, specifically autophagy and endolysosomes, transcription factors, endolysosomal signaling, and mitochondrial homeostasis in shaping cellular bioenergetics. This review aims to summarize the relationship between degradation processes and bioenergetics, providing a foundation for future research to unveil deeper mechanistic insights into bioenergetic regulation.
基金supported by the National Natural Science Foundation(82072729)Wuhan Municipal Health Commission Medical Research Project(WX19Q30),China。
文摘Objective:Oxaliplatin(OXA)and 5-fluorouracil(5-FU)are 2 commonly used chemotherapeutic agents for colorectal cancer(CRC).MicroRNAs(miRNAs,miRs)play crucial roles in the development of chemoresistance in various cancers.However,the role and mechanism of miR-224-5p in regulating CRC chemoresistance remain unclear.This study aims to investigate the function of miR-224-5p in chemoresistant CRC cells and the underlying mechanisms.Methods:CRC datasets GSE28702 and GSE69657 were downloaded from the Gene Expression Omnibus(GEO)database.Differentially expressed miRNAs between drug sensitive and resistant groups(OXA or 5-FU)were analyzed,and miR-224-5p was identified as the target miRNA.Chemoresistant cell lines HCT15-OXR,HCT15-5-FU,SW480-OXR,and SW480-5-FU were established.Transient transfections were performed using miR-224-5p mimics,inhibitors,and their respective negative controls(control mimic,control inhibitor)in these cell lines.Cells were treated with different concentrations of OXA or 5-FU post-transfection,and the half-maximal inhibitory concentration(IC_(50))was determined using the cell counting kit-8(CCK-8)assay.Cell proliferation was assessed by CCK-8 and colony formation assays.The expression levels of miR-224-5p,LC3,and P62 were measured by real-time polymerase chain reaction(real-time PCR)and/or Western blotting.Autophagic flux was assessed using a tandem fluorescent-tagged LC3 reporter assay.TargetScan 8.0,miRTarBase,miRPathDB,and HADb were used to predict B-cell lymphoma-2(Bcl-2)as a potential miR-244-5p target,which was further validated by dual luciferase reporter assays.Results:Chemoresistant CRC cells exhibited down-regulated miR-224-5p expression,whereas up-regulation of miR-224-5p enhanced chemotherapy sensitivity.Exposure to OXA or 5-FU significantly increased autophagic activity in chemoresistant CRC cells,which was reversed by miR-224-5p overexpression.Dual-luciferase assays verified Bcl-2 as a direct target of miR-224-5p.Conclusion:MiR-224-5p regulates chemoresistance in CRC by modulating autophagy through direct targeting of Bcl-2.
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 the STI2030-Major Projects(2021ZD0204001)the National Natural Science Foundation of China(92049120,81870897,81271424,81671111,and 62475179)+8 种基金the Sino German Cooperation and Exchange Project(M-0679)the Guangdong Key Project in the Development of New Tools for the Diagnosis and Treatment of Autism(2018B030335001)the Natural Science Foundation of Jiangsu Province(BK20181436)the Priority Academic Program Development of Jiangsu Higher Education Institutions,the Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases(BM2013003)Suzhou International Joint Laboratory for Diagnosis and Treatment of Brain Diseases,Suzhou Science and Technology Plan Medical and Health Care Science and Technology Innovation Applied Basic Research(SKY2022161)the Research Project of Neurological Diseases in the Second Affiliated Hospital of Soochow University Research Center(ND2023A01)Boxi clinical research project of The First Affiliated Hospital of Soochow University(BXQN202204)Suzhou Science&Technology Projects for People's Livelihood(SKY2021065)Wuxi Municipal Health Commission(M202204).
文摘Oligodendrocyte lineage cells, including oligodendrocyte precursor cells (OPCs) and oligodendrocytes (OLs), are essential in establishing and maintaining brain circuits. Autophagy is a conserved process that keeps the quality of organelles and proteostasis. The role of autophagy in oligodendrocyte lineage cells remains unclear. The present study shows that autophagy is required to maintain the number of OPCs/OLs and myelin integrity during brain aging. Inactivation of autophagy in oligodendrocyte lineage cells increases the number of OPCs/OLs in the developing brain while exaggerating the loss of OPCs/OLs with brain aging. Inactivation of autophagy in oligodendrocyte lineage cells impairs the turnover of myelin basic protein (MBP). It causes MBP to accumulate in the cytoplasm as multimeric aggregates and fails to be incorporated into integral myelin, which is associated with attenuated endocytic recycling. Inactivation of autophagy in oligodendrocyte lineage cells impairs myelin integrity and causes demyelination. Thus, this study shows autophagy is required to maintain myelin quality during aging by controlling the turnover of myelin components.
基金supported by National Natural Science Foundation of China(No.82300268)Guangzhou Municipal Science and Technology Project(No.2023B03J1255).
文摘Objective Electronic cigarettes(ECs)differ from traditional tobacco smoke but may contribute to cardiopulmonary remodeling.Pulmonary hypertension(PH),characterized by pulmonary artery and right ventricle remodeling,poses a significant risk of mortality in infants,children,and adolescents.However,the impact of maternal EC exposure on PH development in offspring remains unclear.To address this,we established a PH rat model with maternal EC exposure.Methods Maternal EC exposure was initiated on gestation day 12 via electronic nicotine delivery systems.Offspring were administered monocrotaline(MCT)at 6 weeks of age(6-wo)to induce PH.Mechanistic experiments were conducted at 10-week-old(10-wo).Protein expression of NADPH oxidases,DNA methyltransferases,and autophagy-related markers was analyzed by Western blot.Morphological changes and the severity of PH were evaluated via hematoxylin and eosin(HE)staining and echocardiography,respectively.Furthermore,the involvement of the oxidative stress/DNA methylation/autophagy axis in response to maternal EC exposure was confirmed through a combination of ELISA,Western blot,HE staining,and echocardiography.Additionally,ATG5 mRNA expression was measured by qRT-PCR.Results Compared with control conditions,maternal EC exposure significantly worsened MCT-induced PH in male offspring.This was associated with increased oxidative stress,DNA hypomethylation,and anomalous autophagy in the offspring.In vivo treatment with chloroquine inhibited autophagy and ameliorated PH development in offspring exposed to maternal EC.Furthermore,N-acetylcysteine(NAC),an antioxidant,attenuated maternal EC exposure-induced oxidative stress,DNA hypomethylation,and excessive autophagy,thereby improving PH.DNA hypermethylation also reversed PH development,accompanied by reduced oxidative stress and suppressed autophagy.ATG5,a key regulator of autophagy,was identified as a potential therapeutic target,as its repression mitigated PH in maternal EC-exposed offspring.Conclusion Maternal EC exposure induces oxidative stress and DNA hypomethylation in offspring,leading to anomalous autophagy and exacerbation of PH development.Targeting ATG5-mediated autophagy may represent a novel therapeutic approach for improving PH outcomes in offspring exposed to maternal EC.Graphical Abstract Pregnant rats were exposed to either EC vapor or standard air from gestation day 12 until 2 days before delivery,with all offspring undergoing PH induction at 6-wo.Offspring exposed to maternal EC presented increased oxidative stress,which in turn affected DNA methylation patterns.The decreased DNA methylation in male offspring led to the activation of autophagy,exacerbating the development of PH.Treatment with ATG5 siRNA inhibited autophagy and alleviated heightened PH in male offspring with maternal EC exposure.
基金supported by the National Natural Science Foundation of China under grant number U21A20259the National Key Research and Development Program of China under grant number 2021YFD1800401.
文摘Mycobacterium tuberculosis(Mtb)employs multiple mechanisms,such as phagocytosis and autophagy,to evade innate immune clearance and establish infection.In the present study,we identified the ESX-1 secretion-associated protein EspL,which promotes Mtb survival by inhibiting phagosome maturation and autophagy initiation.EspL knockout decreased Mtb intracellular survival,while EspL overexpression increased bacterial survival by interfering with phagocytosis and autophagy.EspL interacts with ULK1 and promotes its phosphorylation at Ser^(757),leading to the inhibition of autophagy initiation.Additionally,overexpression of EspL reduced antigen presentation and T-cell responses both in vitro and in vivo.Our findings revealed that EspL interferes with autophagy and antigen presenta-tion by suppressing ULK1 activation.These insights provide a novel understanding of Mtb pathogenicity.
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(Grant Nos.32302642,32272790)the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study(Grant No.SN-ZJU-SIAS-0011)+1 种基金Collaborative Promotion Program of Zhejiang Provincial Agricultural Technology of China(Grant No.2023ZDXT05)the Fundamental Research Funds for the Central Universities(Grant No.226-2022-00122).
文摘Autophagy is a universal cellular process in eukaryotes that plays a critical role in plant growth and stress response.However,the role of autophagy in fruit ripening is largely unknown.Here,we demonstrated that most autophagy-related genes(ATGs)were up-regulated during tomato(Solanum lycopersicum L.)fruit ripening.By using mutants of different autophagy pathway genes(ATG6,ATG10,ATG18a),we revealed that the deficiency of autophagy delayed the ripening of fruit.Compared with wild-type(WT),the production of ethylene was significantly reduced and the accumulation of lycopene was delayed in atg mutants during fruit ripening.We also observed the contents of glucose and fructose were both significantly decreased in atg mutants compared with WT,while the content of organic acids showed the opposite trend.Additionally,the negative regulator of ethylene production,APETALA2a(AP2a),interacted with ATG8 through a specific ATG8-interacting motif(AIM)and could be degraded through the autophagy pathway.These results demonstrate that autophagy plays a critical role in fruit ripening by regulating ethylene production and the accumulation of pigments,sugars and organic acids in tomato.
