SEPS1基因对内毒素所致脓毒症小鼠的肝脏保护作用

Liver protective effect of SEPS1 gene on endotoxin-induced sepsis in mice

目的研究SEPS1基因对内毒素所致脓毒症小鼠的肝脏保护作用。方法30只小鼠随机分为脓毒症组、非特异干扰组、特异干扰组,每组各10只。实验前12h非特异干扰组静脉转染scrambled siRNA,特异干扰组转染SEPS1 siRNA。实验当天3组动物腹腔内注射内毒素10mg/kg制作脓毒症模型。分别于12h、24h留取标本检测血ALT、AST,肝组织细胞因子TNF-α、IL-6含量,肝组织SEPS1蛋白表达及病理学改变。结果脓毒症小鼠肝脏有不同程度的损伤,表现为ALT、AST含量升高,24h时特异干扰组高于脓毒症组(P<0.05);细胞因子IL-6和TNF-α水平明显升高,24h时特异干扰组高于脓毒症组(P<0.05);WesternBlot检测肝组织SEPS1蛋白表达显示:特异干扰组12h时和24h时表达均较脓毒症组减弱。免疫组化法显示:SEPS1蛋白12h和24h时表达均较脓毒症组减弱。病理显示:12h和24h时均为特异干扰组肝细胞病变最严重。结论SEPS1基因表达的下调与脓毒症小鼠肝脏功能损害加重相关,推测SEPS1基因对内毒素攻击所致脓毒症小鼠肝脏具有保护作用。

Objective To investigate the liver protective effects of SEPS1 gene on endotoxin-induced sepsis in mice. Methods Thirty mice were randomly divided into sepsis group,non-specific interference group and specific interference group, 10 mice in each group. 12 h before the experiment,non-specific interference group was transfected scrambled siRNA and specific interference group was transfected with specific SEPS1 siRNA by mouse tail vein. On experimental day,three groups of animals which were injected intraperitoneally with endotoxin 10 mg/kg produced sepsis model. 12 h,24 h after injection,specimens were obtained from blood and tissue to detect blood ALT,AST,liver tissue cytokines TNF-a,IL-6 levels, and SEPS1 protein expression and pathological changes in liver tissue. Results There were different degrees of liver damage in septic mice, manifested as ALT,AST content increased, which in specific interference group was higher than that in sepsis group（P〈0.05）24 h after injection;cytokine IL-6 and TNF-a levels were significantly increased, which in specific interference group was higher than that in sepsis group 24 h after the injection as well （P〈0.05）;Western Blot showed that in liver tissue the expression of SEPS 1 of 12 h and 24 h in specific interference group was lower than that of sepsis group. Immunohistochemistry showed that the SEPS1 protein of 12 h and 24 h expressed less than that of sepsis group. Pathology showed that the damage of liver cells was the most serious in specific interference group both at 12 h and 24 h. Conclusions Down-regulation of SEPS1 gene expression was related to advanced liver function impairment in septic mice, suggesting that SEPS1 gene has a protective effect on endotoxin-induced sepsis in mouse liver.