Elaidic acid(EA)is a typical trans fatty acid(TFA)that emerges during the processing of various fatty foods.In this study,we found that EA induced renal injury with necroptosis.Pretreatment with a reactive oxygen spec...Elaidic acid(EA)is a typical trans fatty acid(TFA)that emerges during the processing of various fatty foods.In this study,we found that EA induced renal injury with necroptosis.Pretreatment with a reactive oxygen species(ROS)inhibitor and a RIPK3 inhibitor alleviated EA-induced necroptosis.The data indicated that EA induced renal necroptosis through ROS/RIPK3/MLKL pathway.In mechanistic studies,we explored how EA induced ROS production.Results indicated that EA caused mitochondrial damage by testing MMP,MFN1,VDAC,and FIS1.Further,EA suppressed mitophagy by testing the levels of LC3,p62,PINK1,Parkin,colocalization of LC3 and Mito-Tracker Red.Mitophagy is a process of selective degradation of damaged mitochondria.A large number of damaged mitochondria couldn't be cleared by mitophagy in time,which increased ROS levels in renal cells.Pretreatment with a mitophagy activator decreased EA-induced ROS levels and mitochondrial damage.Taken together,our data identified that EA induced renal necroptosis by destroying mitochondria and inhibiting mitophagy,thereby activating the ROS/RIPK3/MLKL pathway.展开更多
Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups ...Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups including pigs fed basal diet and pigs fed diet contaminated with 4 mg/kg DON for 21 d.In Exp.2,12 weaned piglets were divided into 2 groups including con-trol piglets and piglets given a gavage of 2 mg/kg body weight(BW)DON.In Exp.3,24 weaned piglets were used in a 2×2 factorial design and the main factors including necrostatin-1(Nec-1)(DMSO or 0.5 mg/kg BW Nec-1)and DON challenge(saline or 2 mg/kg BW DON gavage).On 21 d in Exp.1,or at 6 h post DON gavage in Exp.2 and 3,pigs were killed for blood samples and liver tissues.Liver histology,blood biochemical indicators,and liver inflamma-tion and necroptosis signals were tested.Results Dietary or oral gavage with DON caused liver morphological damage in piglets.Dietary DON led to hepato-cyte damage indicated by increased aspartate transaminase(AST)activity and AST/alanine aminotransferase(ALT)ratio,and DON gavage also caused hepatocyte damage and cholestasis indicated by increased AST and alkaline phosphatase(AKP)activities.Dietary DON caused liver necroptosis indicated by increased protein abundance of total receptor interacting protein kinase 3(t-RIP3)and total mixed lineage kinase domain-like protein(t-MLKL).Moreover,DON gavage increased mRNA expression of interleukin(IL)-6 and IL-1βin liver.DON gavage also induced liver necroptosis demonstrated by increased protein abundance of t-RIP3,phosphorylated-RIP3(p-RIP3),t-MLKL and p-MLKL.However,pretreatment with Nec-1,a specific inhibitor of necroptosis,inhibited liver necroptosis indi-cated by decreased protein expression of t-RIP3,p-RIP3,t-MLKL and p-MLKL.Nec-1 pretreatment reduced liver morphological damage after DON gavage.Pretreatment with Nec-1 also attenuated liver damage induced by DON indicated by decreased activities of AST and AKP.Furthermore,Nec-1 pretreatment inhibited liver mRNA expression of IL-6 and IL-1βafter DON challenge.Conclusions Our data demonstrate for the first time that necroptosis contributes to DON-induced liver injury and inflammation in piglets.展开更多
We previously demonstrated that inhibiting neural stem cells necroptosis enhances functional recovery after spinal cord injury.While exosomes are recognized as playing a pivotal role in neural stem cells exocrine func...We previously demonstrated that inhibiting neural stem cells necroptosis enhances functional recovery after spinal cord injury.While exosomes are recognized as playing a pivotal role in neural stem cells exocrine function,their precise function in spinal cord injury remains unclear.To investigate the role of exosomes generated following neural stem cells necroptosis after spinal cord injury,we conducted singlecell RNA sequencing and validated that neural stem cells originate from ependymal cells and undergo necroptosis in response to spinal cord injury.Subsequently,we established an in vitro necroptosis model using neural stem cells isolated from embryonic mice aged 16-17 days and extracted exosomes.The results showed that necroptosis did not significantly impact the fundamental characteristics or number of exosomes.Transcriptome sequencing of exosomes in necroptosis group identified 108 differentially expressed messenger RNAs,104 long non-coding RNAs,720 circular RNAs,and 14 microRNAs compared with the control group.Construction of a competing endogenous RNA network identified the following hub genes:tuberous sclerosis 2(Tsc2),solute carrier family 16 member 3(Slc16a3),and forkhead box protein P1(Foxp1).Notably,a significant elevation in TSC2 expression was observed in spinal cord tissues following spinal cord injury.TSC2-positive cells were localized around SRY-box transcription factor 2-positive cells within the injury zone.Furthermore,in vitro analysis revealed increased TSC2 expression in exosomal receptor cells compared with other cells.Further assessment of cellular communication following spinal cord injury showed that Tsc2 was involved in ependymal cellular communication at 1 and 3 days post-injury through the epidermal growth factor and midkine signaling pathways.In addition,Slc16a3 participated in cellular communication in ependymal cells at 7 days post-injury via the vascular endothelial growth factor and macrophage migration inhibitory factor signaling pathways.Collectively,these findings confirm that exosomes derived from neural stem cells undergoing necroptosis play an important role in cellular communication after spinal cord injury and induce TSC2 upregulation in recipient cells.展开更多
Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiolog...Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiological factors have been shown to cause acquired SNHL,such as ototoxic drugs,noise exposure,aging,infections,and diseases.Several programmed cell death(PCD)pathways have been reported to be involved in SNHL,especially some novel PCD pathways that have only recently been reported,such as ferroptosis,necroptosis,and pyroptosis.Here we summarize these PCD pathways and their roles and mechanisms in SNHL,aiming to provide new insights and potential therapeutic strategies for SNHL by targeting these PCD pathways.展开更多
Necroptosis,a necrotic form of regulated cell death,plays a crucial role in various tissues and disorders,including sepsis.This process occurs primarily through a caspase-independent mechanism mediated by receptor-int...Necroptosis,a necrotic form of regulated cell death,plays a crucial role in various tissues and disorders,including sepsis.This process occurs primarily through a caspase-independent mechanism mediated by receptor-interacting protein kinase 1(RIPK1),RIPK3,and mixed lineage kinase domain-like(MLKL).Necroptosis-related diseases frequently manifest with excessive inflammatory responses.Corilagin,a gallotannin exhibiting potent anti-inflammatory and anti-oxidant properties,has received increasing attention.However,its effects on necroptosis and associated disorders remain unexplored.In this study,we utilize a surface plasmon resonance-liquid chromatography-tandem mass spectrometry(SPR-LCMS/MS)screening approach to identify corilagin's target proteins and demonstrate its binding to necroptosis-related proteins.In vitro,corilagin inhibits necroptosis induced by either tuberculosis,tumor necrosis factor-α(TNF-α),LCL-161,and inhibitor(IDN-6556)(TSI)(tumor necrosis TNF-αcombined with LCL-161(a Smac mimic)and pan-caspase inhibitor IDN-6556),or lipopolysaccharide(LPS)with IDN-6556.Additionally,it suppresses the phosphorylation of MLKL,RIPK1,and RIPK3,while preventing necrosome formation during necroptotic induction.Corilagin also mitigates the TSI-induced reduction in mitochondrial membrane potential,a characteristic of necroptosis-associated mitochondrial dysfunction and the generation of mitochondrial reactive oxygen species(mt ROS).In a mouse model of sepsis associated with necroptosis,corilagin administration reduces the severity of LPS-induced acute lung injury,correlating with decreased MLKL phosphorylation in lung tissues.These results indicate that corilagin attenuates RIPK1/RIPK3/MLKL signaling,potentially through reducing mt ROS production,thereby inhibiting necroptosis and offering protection against LPS-induced acute lung injury.展开更多
Background In intensive aquaculture systems,the frequent incidence of enteritis reduces production efficiency and results in significant economic losses.Protein feeds account for 40%–60%of aquafeed expenses,and with ...Background In intensive aquaculture systems,the frequent incidence of enteritis reduces production efficiency and results in significant economic losses.Protein feeds account for 40%–60%of aquafeed expenses,and with the growth of intensive aquaculture,demand for fishmeal as a key protein source outstrips supply,driving up prices.This study investigated the therapeutic potential of reducing dietary protein levels by 3%and adding enzymatic cottonseed protein(ECP)in juvenile yellow catfish with dextran sulfate sodium(DSS)-induced enteritis.Methods A total of 1,260 healthy juvenile yellow catfish(Pelteobagrus fulvidraco),with an average body weight of 5.90±0.05 g,were randomly allocated into 7 experimental groups,each with 3 replicates.The fish were fed one of seven diets for 10 weeks:a normal-protein diet(42%;NP)and 6 low-protein diets(39%;LP)supplemented with graded levels of ECP at 0%(ECP0),1%(ECP1),2%(ECP2),3%(ECP3),4%(ECP4),and 5%(ECP5),respectively.Subsequently,48 fish from each group were selected to receive 1 mL of 6%DSS solution.Results Our findings demonstrated that:(1)The DSS+ECP0 group aggravated DSS-induced enteritis in juvenile yellow catfish compared to the DSS+NP group.(2)Dietary supplementation of ECP in LP diets significantly enhanced the enzymatic activity and levels of immunoreactive substances,including LZM,C3,C4,and ACP(P<0.05).Mechanistically,first,ECP supplementation modulated macrophage polarization by inhibiting the M1 phenotype while promoting the M2 phenotype,potentially through the JAK-STAT signaling pathway;second,dietary ECP suppressed the phosphorylation cascade of key necroptosis-related proteins,including RIP1,RIP3,and MLKL,potentially via the NF-κB and MAPK signaling pathways.(3)The DSS+ECP2 group demonstrated comparable or superior efficacy to the DSS+NP group in mitigating DSS-induced intestinal enteritis.Conclusions Our results demonstrated that ECP can alleviate DSS-induced enteritis by regulating macrophage polarization and reducing necroptosis.Furthermore,ECP supplementation effectively counteracted the exacerbation of enteritis caused by dietary protein reduction.These findings highlighted the effectiveness and feasibility of ECP in alleviating enteritis and saving protein.展开更多
BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the rol...BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the role of MLKL’s adenosine triphosphate(ATP)-binding pocket in facilitating necroptosis-independent pathways that may contribute to liver disease progression.By focusing on this mechanism,we seek to identify potential therapeutic targets that can modulate MLKL activity,offering new strategies for the prevention and treatment of liver-related pathologies.AIM To investigate the possibility of using the ATP-binding pocket-associated,necro-ptosis-independent MLKL pathway as a target for liver diseases.METHODS Cell death following necroptosis stimuli was evaluated using cell proliferation assays,flow cytometry,and electron microscopy in various cells.The human liver organoid system was used to evaluate whether the MLKL ATP pocket-binding inhibitor could attenuate inflammation.Additionally,alcoholic and non-alcoholic fatty liver diseases animal models were used to determine whether MLKL ATP pocket inhibitors could attenuate liver injury.RESULTS While an MLKL ATP pocket-binding inhibitor did not prevent necroptosis-induced cell death in RAW 264.7 cells,it did reduce the necroptosis-led expression of CXCL2,ICAM,and VCAM.Notably,MLKL ATP pocket inhibitor diminishes the expression of CXCL2,ICAM,and VCAM by inhibiting the IκB kinase and nuclear factor kappa-B pathways without inducing necroptosis-induced cell death in two-dimensional cell culture as well as the human-derived liver organoid system.Although MLKL ATP-binding inhibitor was ineffective in non-alcoholic fatty liver disease animal models,MLKL ATP-binding inhibitor attenuated hepatic inflammation in the alcoholic liver disease model.CONCLUSION MLKL ATP pocket-binding inhibitor exerted anti-inflammatory effects through the necroptosis-independent MLKL pathway in an animal model of alcoholic liver disease.展开更多
Necroptosis is a newly found type of programmed cell death.It is elicited by death receptor ligands under the condition of apoptotic inhibition,and can be specifically blocked by necrostatin-1,a small-molecule compoun...Necroptosis is a newly found type of programmed cell death.It is elicited by death receptor ligands under the condition of apoptotic inhibition,and can be specifically blocked by necrostatin-1,a small-molecule compound.The pathway of necroptosis starts from the activation of death receptors by death receptor ligands,and is relayed in turn with aggregation and activation of RIP1 and RIP3,activation of energy metabolism-related enzymes including glycogen phosphorylase,glutamate-ammonia ligase as well as glutamate dehydrogenase 1.The process increases the substrates of tricarboxylic acid cycle,enhances the mitochondria respiratory chain,and induces excessive production of OFR.OFR destroys the cellular membranes,resulting in cease of ATP production and leakage of lysoenzymes.Consequently,cell necrosis happens. Necroptosis may be one of the main types of cell necrosis in diseases.Necroptosis and apoptosis convert to each other.Necroptosis may be important to cure of two kinds of diseases.One involves acute critical diseases such as acute ischemia,acute inflammation and acute organ failure,etc.The other includes malignant tumors and virus infections.Prevention from necroptosis is beneficial to the therapy for the former.On the contrary,promotion to necroptosis is beneficial to that for the latter.Collectively,the findings of necroptosis make modulation of necrosis possible.The research on necroptosis will certainly promote our understanding in cell death and disease mechanisms as well as clinical therapy.展开更多
目的:探讨活性氧自由基(ROS)在肾小管上皮细胞necroptosis中的作用。方法:构建肾小管上皮细胞HK-2细胞necroptosis模型,检测其ROS升高程度。并使用NADPH酶抑制剂Apocynin抑制HK-2细胞necroptosis模型中ROS的生成,通过流式细胞计数及检测...目的:探讨活性氧自由基(ROS)在肾小管上皮细胞necroptosis中的作用。方法:构建肾小管上皮细胞HK-2细胞necroptosis模型,检测其ROS升高程度。并使用NADPH酶抑制剂Apocynin抑制HK-2细胞necroptosis模型中ROS的生成,通过流式细胞计数及检测necroptosis的关键蛋白观察HK-2细胞necroptosis的变化。结果:使用肿瘤坏死因子α、苄氧羰酰-缬氨酰-丙氨酰-天冬氨酰-氟甲基酮及抗霉素A成功建立了HK-2细胞necroptosis模型,并观察到HK-2细胞发生necroptosis时ROS显著升高(43.29±2.49 vs 25.90±1.27,P<0.001),而使用necrostatin-1抑制necroptosis后ROS生成受到抑制(35.58±1.08 vs 43.29±2.49,P=0.002)。当对necroptosis模型使用Apocynin干预时,HK-2细胞ROS明显下降(30.71±2.82 vs 43.29±2.49,P<0.001),并且流式细胞计数结果显示坏死细胞比例减少(2.00%±0.30%vs 6.99%±2.79%,P<0.001),同时受体相关蛋白3和混合系列蛋白激酶样结构域的磷酸化水平降低。结论:ROS参与了HK-2细胞的necroptosis,并且通过抑制ROS的生成可减少necroptosis发生,提高损伤状态下HK-2细胞存活率,减轻急性肾小管坏死。展开更多
Some scholars have recently developed the concept of PANoptosis in the study of infectious diseases where pyroptosis,apoptosis and necroptosis act in consort in a multimeric protein complex,PANoptosome.This allows all...Some scholars have recently developed the concept of PANoptosis in the study of infectious diseases where pyroptosis,apoptosis and necroptosis act in consort in a multimeric protein complex,PANoptosome.This allows all the components of PANoptosis to be regulated simultaneously.PANoptosis provides a new way to study the regulation of cell death,in that different types of cell death may be regulated at the same time.To test whether PANoptosis exists in diseases other than infectious diseases,we chose cerebral ischemia/reperfusion injury as the research model,collected articles researching cerebral ischemia/reperfusion from three major databases,obtained the original research data from these articles by bibliometrics,data mining and other methods,then integrated and analyzed these data.We selected papers that investigated at least two of the components of PANoptosis to check its occurrence in ischemia/reperfusion.In the cell model simulating ischemic brain injury,pyroptosis,apoptosis and necroptosis occur together and this phenomenon exists widely in different passage cell lines or primary neurons.Pyroptosis,apoptosis and necroptosis also occurred in rat and mouse models of ischemia/reperfusion injury.This confirms that PANoptosis is observed in ischemic brain injury and indicates that PANoptosis can be a target in the regulation of various central nervous system diseases.展开更多
Ischemic brain injury triggers neuronal cell death by apoptosis via caspase activation and by necroptosis through activation of the receptor-interacting protein kinases （RIPK） associated with the tumor necrosis fact...Ischemic brain injury triggers neuronal cell death by apoptosis via caspase activation and by necroptosis through activation of the receptor-interacting protein kinases （RIPK） associated with the tumor necrosis factor-alpha （TNF-a）/death receptor. Recent evidence shows RIPK inhibitors are neuroprotective and al- leviate ischemic brain injury in a number of animal models, however, most have not yet undergone clinical trials and safety in humans remains in question. Dabrafenib, originally identified as a B-raf inhibitor that is currently used to treat melanoma, was later revealed to be a potent RIPK3 inhibitor at micromolar con- centrations. Here, we investigated whether Dabrafenib would show a similar neuroprotective effect in mice subjected to ischemic brain injury by photothrombosis. Dabrafenib administered intraperitoneally at 10 mg/ kg one hour after photothrombosis-induced focal ischemic injury significantly reduced infarct lesion size in C57BL6 mice the following day, accompanied by a markedly attenuated upregulation of TNF-u. However, subsequent lower doses （5 mg/kg/day） failed to sustain this neuroprotective effect after 4 days. Dabrafenib bl ocked lipopolysaccharides-induced activation of TNF-ct in bone marrow-derived macrophages, suggesting that Dabrafenib may attenuate TNF-ct-induced necroptotic pathway after ischemic brain injury. Since Dab- rafenib is already in clinical use for the treatment of melanoma, it might be repurposed for stroke therapy.展开更多
There are two types of cell death-apoptosis and necrosis. Apoptosis is cell death regulated by cell signaling pathways, while necrosis has until recently been considered a passive mechanism of cell death caused by env...There are two types of cell death-apoptosis and necrosis. Apoptosis is cell death regulated by cell signaling pathways, while necrosis has until recently been considered a passive mechanism of cell death caused by environmental pressures. However, recent studies show that necrosis can also be regulated by specific cell signaling pathways. This mode of death, termed necroptosis, has been found to be related to the occurrence and development of many diseases. We used bibliometrics to analyze the global output of literature on necroptosis in the field of neuroscience published in the period 2007–2019 to identify research hotspots and prospects. We included 145 necroptosisrelated publications and 2239 references published in the Web of Science during 2007–2019. Visualization analysis revealed that the number of publications related to necroptosis has increased year by year, reaching a peak in 2019. China is the country with the largest number of publications. Key word and literature analyses demonstrated that mitochondrial function change, stroke, ischemia/reperfusion and neuroinflammation are likely the research hotspots and future directions of necroptosis research in the nervous system. The relationship between immune response-related factors, damage-associated molecular patterns, pathogen-associated molecular patterns and necroptosis may become a potential research hotspot in the future. Taken together, our findings suggest that although the inherent limitations of bibliometrics may affect the accuracy of the literature-based prediction of research hotspots, the results obtained from the included publications can provide a reference for the study of necroptosis in the field of neuroscience.展开更多
Necroptosis is a non-apoptotic programmed cell death pathway,which causes necrosislike morphologic changes and triggers inflammation in the surrounding tissues.Accumulating evidence has demonstrated that necroptosis i...Necroptosis is a non-apoptotic programmed cell death pathway,which causes necrosislike morphologic changes and triggers inflammation in the surrounding tissues.Accumulating evidence has demonstrated that necroptosis is involved in a number of pathological processes that lead to cardiovascular diseases.However,the exact molecular pathways linking them remain unknown.Herein,this review summarizes the necroptosis-related pathways involved in the development of various cardiovascular diseases,including atherosclerosis,cardiac ischemia-reperfusion injury,cardiac hypertrophy,dilated cardiomyopathy and myocardial infarction,and may shed light on the diagnosis and treatment of these diseases.展开更多
基金supported by National Key Research and Development Program of China(2023YFD1800902).
文摘Elaidic acid(EA)is a typical trans fatty acid(TFA)that emerges during the processing of various fatty foods.In this study,we found that EA induced renal injury with necroptosis.Pretreatment with a reactive oxygen species(ROS)inhibitor and a RIPK3 inhibitor alleviated EA-induced necroptosis.The data indicated that EA induced renal necroptosis through ROS/RIPK3/MLKL pathway.In mechanistic studies,we explored how EA induced ROS production.Results indicated that EA caused mitochondrial damage by testing MMP,MFN1,VDAC,and FIS1.Further,EA suppressed mitophagy by testing the levels of LC3,p62,PINK1,Parkin,colocalization of LC3 and Mito-Tracker Red.Mitophagy is a process of selective degradation of damaged mitochondria.A large number of damaged mitochondria couldn't be cleared by mitophagy in time,which increased ROS levels in renal cells.Pretreatment with a mitophagy activator decreased EA-induced ROS levels and mitochondrial damage.Taken together,our data identified that EA induced renal necroptosis by destroying mitochondria and inhibiting mitophagy,thereby activating the ROS/RIPK3/MLKL pathway.
基金Project of National Natural Science Foundation of China(U22A20517 and 32272906).
文摘Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups including pigs fed basal diet and pigs fed diet contaminated with 4 mg/kg DON for 21 d.In Exp.2,12 weaned piglets were divided into 2 groups including con-trol piglets and piglets given a gavage of 2 mg/kg body weight(BW)DON.In Exp.3,24 weaned piglets were used in a 2×2 factorial design and the main factors including necrostatin-1(Nec-1)(DMSO or 0.5 mg/kg BW Nec-1)and DON challenge(saline or 2 mg/kg BW DON gavage).On 21 d in Exp.1,or at 6 h post DON gavage in Exp.2 and 3,pigs were killed for blood samples and liver tissues.Liver histology,blood biochemical indicators,and liver inflamma-tion and necroptosis signals were tested.Results Dietary or oral gavage with DON caused liver morphological damage in piglets.Dietary DON led to hepato-cyte damage indicated by increased aspartate transaminase(AST)activity and AST/alanine aminotransferase(ALT)ratio,and DON gavage also caused hepatocyte damage and cholestasis indicated by increased AST and alkaline phosphatase(AKP)activities.Dietary DON caused liver necroptosis indicated by increased protein abundance of total receptor interacting protein kinase 3(t-RIP3)and total mixed lineage kinase domain-like protein(t-MLKL).Moreover,DON gavage increased mRNA expression of interleukin(IL)-6 and IL-1βin liver.DON gavage also induced liver necroptosis demonstrated by increased protein abundance of t-RIP3,phosphorylated-RIP3(p-RIP3),t-MLKL and p-MLKL.However,pretreatment with Nec-1,a specific inhibitor of necroptosis,inhibited liver necroptosis indi-cated by decreased protein expression of t-RIP3,p-RIP3,t-MLKL and p-MLKL.Nec-1 pretreatment reduced liver morphological damage after DON gavage.Pretreatment with Nec-1 also attenuated liver damage induced by DON indicated by decreased activities of AST and AKP.Furthermore,Nec-1 pretreatment inhibited liver mRNA expression of IL-6 and IL-1βafter DON challenge.Conclusions Our data demonstrate for the first time that necroptosis contributes to DON-induced liver injury and inflammation in piglets.
基金supported by the National Natural Science Foundation of China,No.81801907(to NC)Shenzhen Key Laboratory of Bone Tissue Repair and Translational Research,No.ZDSYS20230626091402006(to NC)+2 种基金Sanming Project of Medicine in Shenzhen,No.SZSM201911002(to SL)Foundation of Shenzhen Committee for Science and Technology Innovation,Nos.JCYJ20230807110310021(to NC),JCYJ20230807110259002(to JL)Science and Technology Program of Guangzhou,No.2024A04J4716(to TL)。
文摘We previously demonstrated that inhibiting neural stem cells necroptosis enhances functional recovery after spinal cord injury.While exosomes are recognized as playing a pivotal role in neural stem cells exocrine function,their precise function in spinal cord injury remains unclear.To investigate the role of exosomes generated following neural stem cells necroptosis after spinal cord injury,we conducted singlecell RNA sequencing and validated that neural stem cells originate from ependymal cells and undergo necroptosis in response to spinal cord injury.Subsequently,we established an in vitro necroptosis model using neural stem cells isolated from embryonic mice aged 16-17 days and extracted exosomes.The results showed that necroptosis did not significantly impact the fundamental characteristics or number of exosomes.Transcriptome sequencing of exosomes in necroptosis group identified 108 differentially expressed messenger RNAs,104 long non-coding RNAs,720 circular RNAs,and 14 microRNAs compared with the control group.Construction of a competing endogenous RNA network identified the following hub genes:tuberous sclerosis 2(Tsc2),solute carrier family 16 member 3(Slc16a3),and forkhead box protein P1(Foxp1).Notably,a significant elevation in TSC2 expression was observed in spinal cord tissues following spinal cord injury.TSC2-positive cells were localized around SRY-box transcription factor 2-positive cells within the injury zone.Furthermore,in vitro analysis revealed increased TSC2 expression in exosomal receptor cells compared with other cells.Further assessment of cellular communication following spinal cord injury showed that Tsc2 was involved in ependymal cellular communication at 1 and 3 days post-injury through the epidermal growth factor and midkine signaling pathways.In addition,Slc16a3 participated in cellular communication in ependymal cells at 7 days post-injury via the vascular endothelial growth factor and macrophage migration inhibitory factor signaling pathways.Collectively,these findings confirm that exosomes derived from neural stem cells undergoing necroptosis play an important role in cellular communication after spinal cord injury and induce TSC2 upregulation in recipient cells.
基金supported by grants from the National Key Research and Development Program of China(2023YFA1801804,2022YFA0807000,2021YFA1101300,2021YFA1101800,and 2020YFA0112503)the National Natural Science Foundation of China(82171149,82371166,81970892,82330033,82030029,92149304,82071053,and 82171144)+5 种基金the Shenzhen Science and Technology Program(JCYJ20230807114700001,JCYJ20210324125608022,and JCYJ20190814093401920)the Guangdong Basic and Applied Basic Research Foundation(2024A1515010548)the Science and Technology Department of Sichuan Province(2021YFS0371)the Jiangsu Provincial Scientific Research Center of Applied Mathematics(BK20233002)the Open Research Fund of Guangdong Academy of Medical Sciences(YKY‐KF202201)the Taishan Scholars Project-Young Experts Program of Shandong Province(tsqn202211357).
文摘Sensorineural hearing loss(SNHL),the most commonly-occurring form of hearing loss,is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea.Numerous environmental and physiological factors have been shown to cause acquired SNHL,such as ototoxic drugs,noise exposure,aging,infections,and diseases.Several programmed cell death(PCD)pathways have been reported to be involved in SNHL,especially some novel PCD pathways that have only recently been reported,such as ferroptosis,necroptosis,and pyroptosis.Here we summarize these PCD pathways and their roles and mechanisms in SNHL,aiming to provide new insights and potential therapeutic strategies for SNHL by targeting these PCD pathways.
基金supported by the National Natural Science Foundation of China(No.81902634)Guangdong Basic and Applied Basic Research Foundation(No.2024A1515140182)。
文摘Necroptosis,a necrotic form of regulated cell death,plays a crucial role in various tissues and disorders,including sepsis.This process occurs primarily through a caspase-independent mechanism mediated by receptor-interacting protein kinase 1(RIPK1),RIPK3,and mixed lineage kinase domain-like(MLKL).Necroptosis-related diseases frequently manifest with excessive inflammatory responses.Corilagin,a gallotannin exhibiting potent anti-inflammatory and anti-oxidant properties,has received increasing attention.However,its effects on necroptosis and associated disorders remain unexplored.In this study,we utilize a surface plasmon resonance-liquid chromatography-tandem mass spectrometry(SPR-LCMS/MS)screening approach to identify corilagin's target proteins and demonstrate its binding to necroptosis-related proteins.In vitro,corilagin inhibits necroptosis induced by either tuberculosis,tumor necrosis factor-α(TNF-α),LCL-161,and inhibitor(IDN-6556)(TSI)(tumor necrosis TNF-αcombined with LCL-161(a Smac mimic)and pan-caspase inhibitor IDN-6556),or lipopolysaccharide(LPS)with IDN-6556.Additionally,it suppresses the phosphorylation of MLKL,RIPK1,and RIPK3,while preventing necrosome formation during necroptotic induction.Corilagin also mitigates the TSI-induced reduction in mitochondrial membrane potential,a characteristic of necroptosis-associated mitochondrial dysfunction and the generation of mitochondrial reactive oxygen species(mt ROS).In a mouse model of sepsis associated with necroptosis,corilagin administration reduces the severity of LPS-induced acute lung injury,correlating with decreased MLKL phosphorylation in lung tissues.These results indicate that corilagin attenuates RIPK1/RIPK3/MLKL signaling,potentially through reducing mt ROS production,thereby inhibiting necroptosis and offering protection against LPS-induced acute lung injury.
基金National Science Fund for Distinguished Young Scholars of China(32425056)National Natural Science Foundation of China(U23A20250)+2 种基金the earmarked fund for CARS(CARS-45)the National Key R&D Program of China(2023YFD2400600)Sichuan Innovation Team of National Modern Agricultural Industry Technology System(SCCXTD-2024-15).
文摘Background In intensive aquaculture systems,the frequent incidence of enteritis reduces production efficiency and results in significant economic losses.Protein feeds account for 40%–60%of aquafeed expenses,and with the growth of intensive aquaculture,demand for fishmeal as a key protein source outstrips supply,driving up prices.This study investigated the therapeutic potential of reducing dietary protein levels by 3%and adding enzymatic cottonseed protein(ECP)in juvenile yellow catfish with dextran sulfate sodium(DSS)-induced enteritis.Methods A total of 1,260 healthy juvenile yellow catfish(Pelteobagrus fulvidraco),with an average body weight of 5.90±0.05 g,were randomly allocated into 7 experimental groups,each with 3 replicates.The fish were fed one of seven diets for 10 weeks:a normal-protein diet(42%;NP)and 6 low-protein diets(39%;LP)supplemented with graded levels of ECP at 0%(ECP0),1%(ECP1),2%(ECP2),3%(ECP3),4%(ECP4),and 5%(ECP5),respectively.Subsequently,48 fish from each group were selected to receive 1 mL of 6%DSS solution.Results Our findings demonstrated that:(1)The DSS+ECP0 group aggravated DSS-induced enteritis in juvenile yellow catfish compared to the DSS+NP group.(2)Dietary supplementation of ECP in LP diets significantly enhanced the enzymatic activity and levels of immunoreactive substances,including LZM,C3,C4,and ACP(P<0.05).Mechanistically,first,ECP supplementation modulated macrophage polarization by inhibiting the M1 phenotype while promoting the M2 phenotype,potentially through the JAK-STAT signaling pathway;second,dietary ECP suppressed the phosphorylation cascade of key necroptosis-related proteins,including RIP1,RIP3,and MLKL,potentially via the NF-κB and MAPK signaling pathways.(3)The DSS+ECP2 group demonstrated comparable or superior efficacy to the DSS+NP group in mitigating DSS-induced intestinal enteritis.Conclusions Our results demonstrated that ECP can alleviate DSS-induced enteritis by regulating macrophage polarization and reducing necroptosis.Furthermore,ECP supplementation effectively counteracted the exacerbation of enteritis caused by dietary protein reduction.These findings highlighted the effectiveness and feasibility of ECP in alleviating enteritis and saving protein.
基金Supported by the National Research Foundation of Korea Grant Funded by the Korea Government,No.RS-2024-00440477the Korea Institute of Science and Technology Institutional Program,No.2E33111-24-042.
文摘BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the role of MLKL’s adenosine triphosphate(ATP)-binding pocket in facilitating necroptosis-independent pathways that may contribute to liver disease progression.By focusing on this mechanism,we seek to identify potential therapeutic targets that can modulate MLKL activity,offering new strategies for the prevention and treatment of liver-related pathologies.AIM To investigate the possibility of using the ATP-binding pocket-associated,necro-ptosis-independent MLKL pathway as a target for liver diseases.METHODS Cell death following necroptosis stimuli was evaluated using cell proliferation assays,flow cytometry,and electron microscopy in various cells.The human liver organoid system was used to evaluate whether the MLKL ATP pocket-binding inhibitor could attenuate inflammation.Additionally,alcoholic and non-alcoholic fatty liver diseases animal models were used to determine whether MLKL ATP pocket inhibitors could attenuate liver injury.RESULTS While an MLKL ATP pocket-binding inhibitor did not prevent necroptosis-induced cell death in RAW 264.7 cells,it did reduce the necroptosis-led expression of CXCL2,ICAM,and VCAM.Notably,MLKL ATP pocket inhibitor diminishes the expression of CXCL2,ICAM,and VCAM by inhibiting the IκB kinase and nuclear factor kappa-B pathways without inducing necroptosis-induced cell death in two-dimensional cell culture as well as the human-derived liver organoid system.Although MLKL ATP-binding inhibitor was ineffective in non-alcoholic fatty liver disease animal models,MLKL ATP-binding inhibitor attenuated hepatic inflammation in the alcoholic liver disease model.CONCLUSION MLKL ATP pocket-binding inhibitor exerted anti-inflammatory effects through the necroptosis-independent MLKL pathway in an animal model of alcoholic liver disease.
文摘Necroptosis is a newly found type of programmed cell death.It is elicited by death receptor ligands under the condition of apoptotic inhibition,and can be specifically blocked by necrostatin-1,a small-molecule compound.The pathway of necroptosis starts from the activation of death receptors by death receptor ligands,and is relayed in turn with aggregation and activation of RIP1 and RIP3,activation of energy metabolism-related enzymes including glycogen phosphorylase,glutamate-ammonia ligase as well as glutamate dehydrogenase 1.The process increases the substrates of tricarboxylic acid cycle,enhances the mitochondria respiratory chain,and induces excessive production of OFR.OFR destroys the cellular membranes,resulting in cease of ATP production and leakage of lysoenzymes.Consequently,cell necrosis happens. Necroptosis may be one of the main types of cell necrosis in diseases.Necroptosis and apoptosis convert to each other.Necroptosis may be important to cure of two kinds of diseases.One involves acute critical diseases such as acute ischemia,acute inflammation and acute organ failure,etc.The other includes malignant tumors and virus infections.Prevention from necroptosis is beneficial to the therapy for the former.On the contrary,promotion to necroptosis is beneficial to that for the latter.Collectively,the findings of necroptosis make modulation of necrosis possible.The research on necroptosis will certainly promote our understanding in cell death and disease mechanisms as well as clinical therapy.
文摘目的:探讨活性氧自由基(ROS)在肾小管上皮细胞necroptosis中的作用。方法:构建肾小管上皮细胞HK-2细胞necroptosis模型,检测其ROS升高程度。并使用NADPH酶抑制剂Apocynin抑制HK-2细胞necroptosis模型中ROS的生成,通过流式细胞计数及检测necroptosis的关键蛋白观察HK-2细胞necroptosis的变化。结果:使用肿瘤坏死因子α、苄氧羰酰-缬氨酰-丙氨酰-天冬氨酰-氟甲基酮及抗霉素A成功建立了HK-2细胞necroptosis模型,并观察到HK-2细胞发生necroptosis时ROS显著升高(43.29±2.49 vs 25.90±1.27,P<0.001),而使用necrostatin-1抑制necroptosis后ROS生成受到抑制(35.58±1.08 vs 43.29±2.49,P=0.002)。当对necroptosis模型使用Apocynin干预时,HK-2细胞ROS明显下降(30.71±2.82 vs 43.29±2.49,P<0.001),并且流式细胞计数结果显示坏死细胞比例减少(2.00%±0.30%vs 6.99%±2.79%,P<0.001),同时受体相关蛋白3和混合系列蛋白激酶样结构域的磷酸化水平降低。结论:ROS参与了HK-2细胞的necroptosis,并且通过抑制ROS的生成可减少necroptosis发生,提高损伤状态下HK-2细胞存活率,减轻急性肾小管坏死。
基金supported by the National Natural Science Foundation of China,Nos.81772134(to KX),81971891(to KX),82172196(to KX),81571939(to KX)the Fundamental Research Funds for the Central Universities of Central South University of China,No.2020zzts218,(to WTY)Hunan Provincial Innovation Foundation For Postgraduate of China,Nos.CX20200116(to WTY),CX20190139(to LSL).
文摘Some scholars have recently developed the concept of PANoptosis in the study of infectious diseases where pyroptosis,apoptosis and necroptosis act in consort in a multimeric protein complex,PANoptosome.This allows all the components of PANoptosis to be regulated simultaneously.PANoptosis provides a new way to study the regulation of cell death,in that different types of cell death may be regulated at the same time.To test whether PANoptosis exists in diseases other than infectious diseases,we chose cerebral ischemia/reperfusion injury as the research model,collected articles researching cerebral ischemia/reperfusion from three major databases,obtained the original research data from these articles by bibliometrics,data mining and other methods,then integrated and analyzed these data.We selected papers that investigated at least two of the components of PANoptosis to check its occurrence in ischemia/reperfusion.In the cell model simulating ischemic brain injury,pyroptosis,apoptosis and necroptosis occur together and this phenomenon exists widely in different passage cell lines or primary neurons.Pyroptosis,apoptosis and necroptosis also occurred in rat and mouse models of ischemia/reperfusion injury.This confirms that PANoptosis is observed in ischemic brain injury and indicates that PANoptosis can be a target in the regulation of various central nervous system diseases.
基金supported by grants from the Heart and Stroke Foundation of Canada(HHC,AFRS)the Canadian Institutes of Health Research(to HHC and AFRS)supported by a Mid-Career Investigator Award from the Heart and Stroke Foundation of Ontario
文摘Ischemic brain injury triggers neuronal cell death by apoptosis via caspase activation and by necroptosis through activation of the receptor-interacting protein kinases （RIPK） associated with the tumor necrosis factor-alpha （TNF-a）/death receptor. Recent evidence shows RIPK inhibitors are neuroprotective and al- leviate ischemic brain injury in a number of animal models, however, most have not yet undergone clinical trials and safety in humans remains in question. Dabrafenib, originally identified as a B-raf inhibitor that is currently used to treat melanoma, was later revealed to be a potent RIPK3 inhibitor at micromolar con- centrations. Here, we investigated whether Dabrafenib would show a similar neuroprotective effect in mice subjected to ischemic brain injury by photothrombosis. Dabrafenib administered intraperitoneally at 10 mg/ kg one hour after photothrombosis-induced focal ischemic injury significantly reduced infarct lesion size in C57BL6 mice the following day, accompanied by a markedly attenuated upregulation of TNF-u. However, subsequent lower doses （5 mg/kg/day） failed to sustain this neuroprotective effect after 4 days. Dabrafenib bl ocked lipopolysaccharides-induced activation of TNF-ct in bone marrow-derived macrophages, suggesting that Dabrafenib may attenuate TNF-ct-induced necroptotic pathway after ischemic brain injury. Since Dab- rafenib is already in clinical use for the treatment of melanoma, it might be repurposed for stroke therapy.
基金supported by the National Natural Science Foundation of China,Nos. 81772134,81971891,and 81571939 (to KX)the Key Research and Development Program of Hunan Province of China,No. 2018SK2091 (to KX)+3 种基金Hunan Provincial Innovation Foundation For Postgraduate,No. CX20200116 (to WTY)Wu Jie Ping Medical Foundation of the Minister of Health of China,No. 320.6750.14118 (to KX)Foundation of Science and Technology of Hunan Province of China,No. 2018JJ2552 (to YC)the Project of Graduate Independent Exploration and Innovation Plan of Central South University of China,No. 2020zzts218 (to WTY)。
文摘There are two types of cell death-apoptosis and necrosis. Apoptosis is cell death regulated by cell signaling pathways, while necrosis has until recently been considered a passive mechanism of cell death caused by environmental pressures. However, recent studies show that necrosis can also be regulated by specific cell signaling pathways. This mode of death, termed necroptosis, has been found to be related to the occurrence and development of many diseases. We used bibliometrics to analyze the global output of literature on necroptosis in the field of neuroscience published in the period 2007–2019 to identify research hotspots and prospects. We included 145 necroptosisrelated publications and 2239 references published in the Web of Science during 2007–2019. Visualization analysis revealed that the number of publications related to necroptosis has increased year by year, reaching a peak in 2019. China is the country with the largest number of publications. Key word and literature analyses demonstrated that mitochondrial function change, stroke, ischemia/reperfusion and neuroinflammation are likely the research hotspots and future directions of necroptosis research in the nervous system. The relationship between immune response-related factors, damage-associated molecular patterns, pathogen-associated molecular patterns and necroptosis may become a potential research hotspot in the future. Taken together, our findings suggest that although the inherent limitations of bibliometrics may affect the accuracy of the literature-based prediction of research hotspots, the results obtained from the included publications can provide a reference for the study of necroptosis in the field of neuroscience.
基金This work was supported by grants from the Natural Science Foundation of Jiangxi Province (No.20161bab215222Educational Commission of Jiangxi Province of China (No.gjjl50147)Cultivation Scientific Research Fund for the Junior Teachers of Medicine in Nanchang University (No.py201826).
文摘Necroptosis is a non-apoptotic programmed cell death pathway,which causes necrosislike morphologic changes and triggers inflammation in the surrounding tissues.Accumulating evidence has demonstrated that necroptosis is involved in a number of pathological processes that lead to cardiovascular diseases.However,the exact molecular pathways linking them remain unknown.Herein,this review summarizes the necroptosis-related pathways involved in the development of various cardiovascular diseases,including atherosclerosis,cardiac ischemia-reperfusion injury,cardiac hypertrophy,dilated cardiomyopathy and myocardial infarction,and may shed light on the diagnosis and treatment of these diseases.
