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.
目的:通过盲肠穿刺结扎法建立脓毒症大鼠模型,观察四逆汤是否可以改善脓毒症SD大鼠肾损伤,探讨四逆汤是否通过抑制NLRP3-Caspase-1-IL-1β/IL-18信号通路改善脓毒症SD大鼠肾损伤。方法:100只雌性大鼠,按照大鼠体重进行初步分类,采用轮次分配法,确保每组大鼠体重分布均匀。平均分成3大组,每组30只老鼠,从中随机选取1组作为空白组,剩下两组使用CLP法进行造模,分别是实验组、对照组两个大组。随后又分别按大鼠体重采用轮次分类分为8小时组、16小时组和24小时组三个小组,每个小组10只;其中死亡10只,继续造模补充;实验组采用四逆汤灌胃、对照组和空白组采用生理盐水灌胃。分别在8 h、16 h、24 h时间点,取各组大鼠肾组织进行HE染色,计算肾损伤评分,WB检测肾组织中NLRP3、Caspase-1蛋白表达,ELISA法检测血清IL-1β、IL-18表达情况。结果:1) 肾组织HE染色后高倍镜下对照组8小时开始可见肾脏正常形态被破坏,肾小球体积明显增大,肾小囊腔扩大,系膜细胞肿胀、间质炎性细胞浸润;16小时较8小时更加严重,并且间质有明显出血及大量炎症细胞浸润;24小时肾小球缺血皱缩,失去正常形态;球–囊间隙扩大,炎性细胞渗出增多,肾小管上皮细胞肿胀。实验组大鼠8小时及16小时的肾脏病理改变较对照组明显减轻。肾损伤评分中,肾损伤随着时间的延长而逐渐加重(P P P P > 0.05)。3) ELISA检测血清IL-1β、IL-18后发现,总趋势是从8小时开始明显上升(P P P > 0.05)。这与WB检查结果相似。结论:四逆汤可能通过介导NLRP3-Caspase-1-IL-1β/IL-18信号通路改善脓毒症急性肾损伤SD大鼠肾损伤。Objective: Sepsis rat model was established by cecal puncture and ligation to observe whether Sini Decoction could improve kidney injury of septic SD rats, and to explore whether Sini Decoction could improve kidney injury of septic SD rats by inhibiting NLRp3-caspase-1-IL-1β/I
