Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that ...Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.展开更多
Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI....Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI.However,the mechanism remains unclear.This study aimed to investigate whether c-Myc affects CIRI by regulating Serine/Arginine-rich Splicing Factor 1(SRSF1)/NLRP1-mediated pyroptosis.Methods:Oxygen-glucose deprivation/reperfusion(OGD/R)induced neuroblastoma cells for the establishment of an in vitro CIRI model.The levels of c-Myc and SRSF1,cell viability,the expression of pyroptosis-related factors,and the interaction between SRSF1 and NLRP1 were evaluated.Results:The expression of c-Myc and SRSF1 was decreased in OGD/R-induced neuroblastoma cells.c-Myc overexpression increased c-Myc and SRSF1 expression and cell viability in OGD/Rinduced neuroblastoma cells while inhibiting NLRP1,Caspase1,apoptosis-associated speck-like protein containing a CARD(ASC),interleukin-1beta(IL-1β),IL-18,and lactate dehydrogenase levels and pyroptosis.C-Myc was positively correlated with SRSF1.SRSF1 low expression reversed the effects of c-Myc on the above indicators in OGD/Rinduced neuroblastoma cells.Mechanically,SRSF1 interacted with NLRP1.SRSF1 was negatively correlated with NLRP1.The NLRP1 activator muramyl dipeptide(MDP)reversed the SRSF1 effect on OGD/R-induced neuroblastoma cells.Conclusion:Our results indicated that c-Myc reduced OGD/R-induced neuroblastoma cell pyroptosis by inhibiting NLRP1 activation by positive feedback SRSF1 signal.Our findings suggested that the c-Myc/SRSF1 axis might be a new strategy for treating CIRI in the clinic.展开更多
炎性复合体是存在于胞浆中的一组多蛋白复合体,它能够活化胱天蛋白酶(caspase)-1,后者介导IL(interleukin)-1β、IL-18和IL-33等促炎因子的成熟与释放。NALP1(NACHT leucine-rich-repeat protein 1)也称NLRP1,是最早被鉴定出来的具有明...炎性复合体是存在于胞浆中的一组多蛋白复合体,它能够活化胱天蛋白酶(caspase)-1,后者介导IL(interleukin)-1β、IL-18和IL-33等促炎因子的成熟与释放。NALP1(NACHT leucine-rich-repeat protein 1)也称NLRP1,是最早被鉴定出来的具有明确配体的炎性复合体之一,它参与多种炎症反应和细胞凋亡的调节作用。此外,还有研究发现NLRP1在急性白血病的发生发展及诱导骨髓造血干细胞凋亡等血液系统疾病中也发挥着重要作用。本文将对NLRP1的结构、活化机制、调控及在造血系统中的作用进行综述。展开更多
文摘Objective Stroke is a leading cause of death and disability worldwide,with ischemic stroke accounting for 80%-85%of cases.Despite the prevalence,effective treatments remain scarce.The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis,contributing to a range of neurocellular dysfunctions.Pyroptosis,a recently identified form of programmed cell death,is characterized by caspase-1 activation and the action of the Gasdermin D(GSDMD)protein family,leading to cell perforation and inflammatory death.Methods In this study,human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide(H_(2)S)against this toxicity through the antagonization of pyroptosis.We employed CCK-8 and LDH assays to assess cell viability.YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability.Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells,that are mitigated by the H_(2)S donor NaHS.Furthermore,ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1(NLRP-1),caspase-1,and the cleavage of GSDMD,with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1,caspase-1,and GSDMD pathway.
基金supported by a fund from the Hainan Provincial Natural Science Foundation of China(No.821MS156).
文摘Objectives:NOD-like receptor family pyrin domain-containing(NLRP)1-mediated pyroptosis plays a key role in the pathogenesis of cerebral ischemia-reperfusion injury(CIRI).C-Myc is reported to play a major role in CIRI.However,the mechanism remains unclear.This study aimed to investigate whether c-Myc affects CIRI by regulating Serine/Arginine-rich Splicing Factor 1(SRSF1)/NLRP1-mediated pyroptosis.Methods:Oxygen-glucose deprivation/reperfusion(OGD/R)induced neuroblastoma cells for the establishment of an in vitro CIRI model.The levels of c-Myc and SRSF1,cell viability,the expression of pyroptosis-related factors,and the interaction between SRSF1 and NLRP1 were evaluated.Results:The expression of c-Myc and SRSF1 was decreased in OGD/R-induced neuroblastoma cells.c-Myc overexpression increased c-Myc and SRSF1 expression and cell viability in OGD/Rinduced neuroblastoma cells while inhibiting NLRP1,Caspase1,apoptosis-associated speck-like protein containing a CARD(ASC),interleukin-1beta(IL-1β),IL-18,and lactate dehydrogenase levels and pyroptosis.C-Myc was positively correlated with SRSF1.SRSF1 low expression reversed the effects of c-Myc on the above indicators in OGD/Rinduced neuroblastoma cells.Mechanically,SRSF1 interacted with NLRP1.SRSF1 was negatively correlated with NLRP1.The NLRP1 activator muramyl dipeptide(MDP)reversed the SRSF1 effect on OGD/R-induced neuroblastoma cells.Conclusion:Our results indicated that c-Myc reduced OGD/R-induced neuroblastoma cell pyroptosis by inhibiting NLRP1 activation by positive feedback SRSF1 signal.Our findings suggested that the c-Myc/SRSF1 axis might be a new strategy for treating CIRI in the clinic.
文摘炎性复合体是存在于胞浆中的一组多蛋白复合体,它能够活化胱天蛋白酶(caspase)-1,后者介导IL(interleukin)-1β、IL-18和IL-33等促炎因子的成熟与释放。NALP1(NACHT leucine-rich-repeat protein 1)也称NLRP1,是最早被鉴定出来的具有明确配体的炎性复合体之一,它参与多种炎症反应和细胞凋亡的调节作用。此外,还有研究发现NLRP1在急性白血病的发生发展及诱导骨髓造血干细胞凋亡等血液系统疾病中也发挥着重要作用。本文将对NLRP1的结构、活化机制、调控及在造血系统中的作用进行综述。
