目的:探讨乌司他丁( ulinastatin,UTI)预处理对大鼠肝脏缺血再灌注(ischemia reperfusion,IR)损伤后炎性反应的影响,并研究高迁移率蛋白B1 (high mobility group box 1,HMGB1)在UTI预处理机制中的作用.方法:采用SD大鼠70％肝脏IR损伤模型,即在肝脏缺血45 min后再灌注2h.将40只大鼠随机为4组,分别为0.9％氯化钠对照组(对照组,n=10)、UTI预处理组(UTI组,n=10)、UTI+ HMGB1拮抗剂组(UTI+ Anti-HMGB1组,n=10)和乌司他丁+HMGB1诱导剂组(UTI+ rHMGB1组,n=10).比较各组大鼠在IR后血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)的水平；采用酶联免疫吸附试验检测各组大鼠血清中肿瘤坏死因子α(tumor necrosis factor-alpha,TNFα)和白细胞介素1(intcrleukin 1,IL1)的水平；对各组大鼠的肝组织进行HE染色观察其病理学改变；采用免疫组织化学染色方法检测各组大鼠肝组织中HMGB1蛋白的表达水平.结果:与对照组相比,UTI组在IR后血清中ALT、AST均显著降低(P＜0.05).UTI+ rHMGB1组IR后血清TNFα、IL1水平与对照组相比无显著差异(P＞0.05)；UTI组和UTI+ Anti-HMGB1组IR后血清TNFα、IL1水平与对照组相比均显著降低(P＜0.05).肝组织病理显示,UTI组和UTI+ Anti-HMGB1组的肝细胞坏死和中性粒细胞浸润显著轻于对照组和UTI+ rHMGB1组.免疫组织化学染色显示,UTI组和UTI+ Anti HMGB1组肝细胞HMGB1表达程度显著低于对照组(P＜0.05).结论:UTI预处理可以显著抑制大鼠肝IR损伤后肝细胞中HMGB1的表达水平以及下游炎症因子的水平,从而减轻IR损伤；HMGB1诱导剂可逆转UTI的肝保护作用.%Objective:To investigate the effect of pretreatment with Ulinastatin(UTI) on the inflammatory reaction induced by hepatic ischemia reperfusion(IR) in rats and the role of high mobility group box 1 (HMGB1) . Methods;SD rat models of hepatic IR (45 minutes of partial ischemia and 2 hours of reperfusion) were used in this study. Forty rats were randomly divided into four groups: 0. 9% saline group(control group),UTI group,UTI + anti-HMGBl group and UTI + rHMGBl group. Blood and hepatic tissue samples have been harvested after reperfusion. Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) have been detected. Serum levels of tumor necrosis factor-α(TNFα) and interleukin-1 (IL1) have also been detected. HE staining have been done to observe the pathological changes of the liver tissues in each group . The expression of HMGB1 in hepatic tissues was assessed by immunohistochemical staining. Results: Serum levels of ALT,AST,TNFα and IL1 in UTI group and UTI + anti-HMGBl group were significantly lower than those in control group and UTI+ rHMGBl group (F<0. 05). There was no significant difference in the serum levels of TNFa and IL1 between control group and UTI + rHMGBl group (P>0. 05). Hepatic structures were better maintained and apoptotic hepatic cells were less in UTI group and UTI + anti-HMGBl group,compared with control group and UTI + rHMGBl group(P<0. 05) . Levels of HMGBl in UTI group and UTI+ anti-HMGBl groups were significantly lower than those in control group(P<0. 05). Conclusion: Pretreatment with UTI can reduce the expression of HMGBl and thus attenuate liver injury through its anti-inflammatory effects. The effects of UTI can be turned over by rHMGBl.
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