异氟醚预处理通过抑制caspase-11相关的非经典细胞焦亡途径减轻小鼠肝脏缺血再灌注损伤

Isoflurane preconditioning inhibits caspase-11-related noncanonical pyroptosis pathway to alleviate hepatic ischemia-reperfusion injury in mice

目的研究异氟醚预处理通过抑制用随机数字表法肝脏缺血再灌注损伤(HIRI)过程中细胞焦亡非经典途径而对HIRI产生保护作用。方法建立肝脏缺血再灌注损伤小鼠模型,将30只C57BL/6小鼠分成假手术组(n=10)、异氟醚组(n=10)和手术组(n=10),其中异氟醚组经1.4%异氟醚预处理30 min后行肝脏缺血再灌注手术。用组织HE染色切片观察及病理评分和检测血清中ALT/AST水平的方法评估异氟醚预处理对肝脏缺血再灌注损伤的保护作用。用ELISA法检测血清中IL-1β和IL-18,用Western Blot法检测肝脏组织中GSDMD的蛋白表达,以评价异氟醚预处理对细胞焦亡的抑制作用。用Western Blot法和免疫荧光法检测肝脏组织中caspase-11的蛋白表达,评价异氟醚预处理对细胞焦亡非经典途径的抑制作用。结果HE染色切片的Suzuki's评分结果显示手术组分数较假手术组显著增高(P<0.05),而异氟醚组的分数较手术组显著降低(P<0.05);ALT/AST检测结果显示假手术组血清中ALT/AST显著增高(P<0.05),而异氟醚组的检测结果较手术组显著降低(P<0.05);ELISA结果提示血清中IL-1β与IL-18在手术组中表达明显高于假手术组(P<0.05),异氟醚组中IL-1β与IL-18均明显低于手术组(P<0.05)。Western blot检测肝组织中手术组GSDMD的表达明显高于假手术组,而异氟醚组的表达较手术组明显降低(P<0.05);Western blot和免疫荧光结果均提示I/R组小鼠肝脏组织中caspase-11表达显著高于假手术组(P<0.05),而异氟醚组小鼠肝脏组织中的caspase-11表达较手术组明显降低(P<0.05)。结论异氟醚预处理对肝脏缺血再灌注损伤有保护作用,而这种作用与其对细胞焦亡非经典途径的抑制相关。

Objective To study the protective effect of isoflurane preconditioning on hepatic ischemia-reperfusion(I/R)injury mediated by the noncanonical pyroptosis pathway.Methods Thirty C57BL/6 mice were randomly divided into sham-operated group,isoflurane group and I/R group,and in the latter two groups,hepatic I/R injury was induced by clamping the portal vein for 30 min.In isoflurane group,the mice were pretreated with 1.4%isoflurane 30 min before the surgery.The protective effect of isoflurane preconditioning against hepatic I/R injury was evaluated by assessing the pathological score of HE staining of the liver tissue and serum ALT and AST levels.Serum IL-1βand IL-18 levels and the protein expression of GSDMS were detected by ELISA and Western blotting to evaluate the inhibitory effect of isoflurane preconditioning on pyroptosis.Western blotting and immunofluroescence were used to detect the protein expression of caspase-11 in the liver tissues to evaluate the inhibitory effect of isoflurane preconditioning on noncanonical pyroptosis pathway.Results The Suzuki's score of the liver tissue was significantly higher in I/R group than in the sham group(P<0.05),while the score in the isoflurane group was significantly lower than that in the I/R group(P<0.05).Serum ALT and AST levels significantly increased in the sham group(P<0.05),and were significantly lower in isoflurane group than in I/R group(P<0.05).The serum levels of IL-1βand IL-18 were significantly higher in I/R group than in sham group(P<0.05),and were significantly lower in isoflurane group than in I/R group(P<0.05).The expression of GSDMD in the I/R group was significantly higher than that in sham group,and was significantly lower in isoflurane group than in I/R group(P<0.05).The hepatic expression of caspase-11 was significantly higher in I/R group than in sham group(P<0.05),and was significantly lower in isoflurane group than in I/R group(P<0.05).Conclusion Isoflurane preconditioning has protective effect against hepatic I/R injury,which is related to the inhibition of the noncanonical pyroptosis pathway.