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论著:乌司他丁对大鼠肝脏缺血再灌注损伤后炎性反应的影响
Ulinastatin Attenuates the Inflammatory Reaction Induced by Hepatic Ischemia Reperfusion in Rats
陈伟新 杨立群
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作者单位:复旦大学附属中山医院青浦分院普外科
中文关键字:乌司他丁;肝脏;缺血再灌注;高迁移率蛋白B1
英文关键字:Ulinastatin; Hepatic;Ischemia reperfusion;High mobility group box 1
中文摘要: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(interleukin 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。
英文摘要: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 HMGB1 group and UTI+ rHMGB1 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 HMGB1 group were significantly lower than those in control group and UTI+rHMGB1 group (P<0.05). There was no significant difference in the serum levels of TNFα and IL1 between control group and UTI+rHMGB1 group (P>0.05). Hepatic structures were better maintained and apoptotic hepatic cells were less in UTI group and UTI +anti HMGB1 group, compared with control group and UTI+rHMGB1 group(P<0.05) . Levels of HMGB1 in UTI group and UTI+anti HMGB1 groups were significantly lower than those in control group(P<0.05). Conclusion: Pretreatment with UTI can reduce the expression of HMGB1 and thus attenuate liver injury through its anti inflammatory effects. The effects of UTI can be turned over by rHMGB1.
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