瑞芬太尼预处理对大鼠肝脏缺血再灌注损伤的影响及机制

Protective effect of remifentanil preconditioning against hepatic ischemia-reperfusion injury in rats: role of p38 mitogen-activated protein kinases

目的研究瑞芬太尼预处理对大鼠肝脏缺血再灌注损伤的保护作用及其可能的作用机制。方法健康SD大鼠96只,随机分为Sham组(S组)和缺血组,缺血组包括缺血再灌注组(I/R组)、瑞芬太尼预处理组(RPC组)、p38丝裂原活化蛋白激酶(p38MAPK)阻滞剂SB203580(SB)+RPC组。各组分别于再灌注末抽取静脉血、处死大鼠,收集肝组织。观察血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)水平;ELISA法检测血清肿瘤坏死因子α(TNF-α)及IL-1β水平;TUNEL法测定肝细胞凋亡情况;HE染色观察肝组织病理学改变;Western blotting法测定肝组织p38MAPK及其磷酸化组分pp38MAPK的表达水平。结果与S组相比,I/R组血清ALT、AST、TNF-α及IL-1β水平均明显升高,出现明显肝组织损伤,肝细胞凋亡增加;与I/R组相比,RPC组血清ALT、AST、TNF-α及IL-1β水平均明显下降,组织病理学损伤明显减轻,肝细胞凋亡减少,肝组织pp38MAPK表达明显升高;应用p38MAPK阻断剂SB203580,肝损伤加重。结论瑞芬太尼预处理通过激活p38MAPK通路,使p38MAPK磷酸化,减轻炎症因子的产生,对大鼠肝脏缺血再灌注损伤起保护作用。这种保护作用可以被SB 203580阻断。

Objective To assess the role of p38 mitogen-activated protein kinases（p38MAPK） in the protective effect of remifentanil preconditioning（RPC） on hepatic ischemia-reperfusion injury in rats.Methods Ninety-six male SD rats were randomly assigned into sham-operated group,ischemia-reperfusion group（I/R group）,RPC group,and SB（an inhibitor of p38 MAPK） ＋RPC group.The rats were sacrificed at the end of reperfusion,and serum levels of alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,tumor necrosis factor-α（TNF-α）,and interleukin-1β（IL-1β） were measured.HE staining was used to observe the hepatic histopathological changes,and Western blotting was employed to examine p38MAPK and pp38MAPK protein expression.TUNEL staining was used to examine cell apoptosis in the liver tissues.Results Compared with sham-operated group,I/R group showed significantly increased serum levels of AST,ALT,TNF-α and IL-1β with obvious histopathological changes and cell apoptosis in the liver.RPC significantly decresed the elevated serum levels of AST,ALT,TNF-α and IL-1β and lessened hepatic histopathological changes,and caused reduced p38MAPK phosphorylation and hepatic cell apoptosis index.The protective effects of RPC were abolished by SB 203580 pretreatment.Conclusion RPC attenuates the production of inflammatory factors by activating p38MAPK signal pathway to improve hepatic ischemia-reperfusion injury,and these effects can be blocked by SB203580,a p38MAPK inhibitor.