七氟醚后处理对缺氧复氧诱导的H9C2心肌细胞焦亡及TXNIP/NLRP3通路的影响

Effects of sevoflurane post-conditioning on hypoxia/reoxygenation-induced pyrolysis of H9C2 cardiomyocytes and TXNIP/NLRP3 pathway

目的:基于硫氧还蛋白结合蛋白(TXNIP)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)通路探究七氟醚后处理(SPostC)对缺氧复氧(HR)诱导的H9C2心肌细胞焦亡的影响。方法:体外培养H9C2心肌细胞,将细胞分为4组:正常对照组(NC组)、HR组、HR+SPostC组、HR+SPostC+TXNIP/NLRP3通路抑制剂白藜芦醇组(HR+SPostC+RES组)。采用CCK-8检测细胞活力;采用乳酸脱氢酶(LDH)试剂盒检测LDH含量;采用碘化丙啶(PI)染色法检测细胞膜孔的形成;采用Western blot法检测细胞焦亡相关蛋白Caspase-1、IL-1β、IL-18及TXNIP/NLRP3通路相关蛋白TXNIP、NLRP3的表达;采用实时荧光定量PCR法检测TXNIP/NLRP3通路相关基因TXNIP、NLRP3的表达。结果:与NC组相比,其余3组细胞活力显著降低,而LDH释放量、PI染色阳性率以及Caspase-1、IL-1β、IL-18、TXNIP和NLRP3的表达明显升高(P<0.05);与HR组相比,HR+SPostC组和HR+SPostC+RES组细胞活力显著升高(P<0.05),而LDH释放量、PI染色阳性率以及Caspase-1、IL-1β、IL-18、TXNIP和NLRP3的表达明显降低(P<0.05);与HR+SPostC组相比,HR+SPostC+RES组细胞活力显著升高(P<0.05),而LDH释放量、PI染色阳性率以及Caspase-1、IL-1β、IL-18、TXNIP和NLRP3的表达明显降低(P<0.05)。结论:SPostC可能通过抑制TXNIP/NLRP3信号通路进而抑制HR诱导的H9C2心肌细胞焦亡。

Objective:Based on the thioredoxin interacting protein(TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway,to investigate the effect of sevoflurane post-conditioning(SPostC)on the pyrolysis of H9C2 cardiomyocytes induced by hypoxia-reoxygenation(HR).Methods:H9C2 cardiomyocytes were cultured in vitro and divided into four groups:normal control group(NC group),HR group,HR+SPostC group,HR+SPostC+TXNIP/NLRP3 pathway inhibitor resveratrol group(HR+SPostC+RES group).Cell viability was detected by CCK-8.Lactate dehydrogenase(LDH)kit was used to detect the content of LDH.Propidium iodide(PI)staining was used to detect the formation of cell membrane pores.Western blot was used to detect the expressions of pyrolysis-related proteins Caspase-1,IL-1β,IL-18 and TXNIP/NLRP3 pathway-related proteins TXNIP and NLRP3.Real-time fluorescent quantitative PCR was used to detect the expressions of TXNIP/NLRP3 pathway related genes TXNIP and NLRP3.Results:Compared with NC group,the cell viability of the other three groups were significantly reduced,the release of LDH,the positive rate of PI staining,and the expressions of Caspase-1,IL-1β,IL-18,TXNIP and NLRP3 were significantly increased(P<0.05).Compared with HR group,the cell viability was significantly increased in tHR+SPostC group and HR+SPostC+RES group,the release of LDH,the positive rate of PI staining and the expressions of Caspase-1,IL-1β,IL-18,TXNIP and NLRP3 were significantly reduced(P<0.05).Compared with HR+SPostC group,in HR+SPostC+RES group,the cell viability was significantly increased,the release of LDH,the positive rate of PI staining,and the expressions of Caspase-1,IL-1β,IL-18,TXNIP and NLRP3 were significantly reduced(P<0.05).Conclusion:SPostC may inhibit HR-induced pyrolysis of H9C2 cardiomyocytes by inhibiting the TXNIP/NLRP3 signaling pathway.