消退素D1预处理对肝缺血再灌注损伤大鼠模型的保护作用及机制

Protective effect of resolvin D1 pretreatment in a rat model of hepatic ischemia-reperfusion injury and its mechanism

目的探究消退素D1(RvD1)对肝缺血再灌注(IR)损伤大鼠模型的保护作用及与血红素氧合酶-1(HO-1)之间的关系。方法Sprague-Dawley大鼠36只随机分为6组,分别为假手术(sham)+PBS组、sham+RvD1高剂量(10μg/kg)组、IR+PBS组、IR+RvD1(2μg/kg)低剂量组、IR+RvD1(5μg/kg)中剂量组和IR+RvD1(10μg/kg)高剂量组,每组6只。RvD1于缺血前1 h腹腔注射。生化仪测定ALT、AST水平,酶联免疫法检测血浆TNFα、IL-6、IL-8水平,HE染色观察肝组织学变化,Western Blot方法检测肝组织HO-1变化。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与IR+PBS组相比,IR+RvD1中剂量组和IR+RvD1高剂量组大鼠ALT、AST水平以及炎症因子TNFα、IL-6、IL-8水平均明显降低(P值均<0.05),且中、高剂量两组间比较差异均无统计学意义(P值均>0.05)。Western Blot结果显示IR+RvD1中剂量组和IR+RvD1高剂量组肝脏HO-1蛋白表达较IR+PBS组升高(P值均<0.05)。HE染色观察肝组织学变化显示,与IR+PBS组相比,IR+RvD1中剂量组和IR+RvD1高剂量组细胞肿胀及肝索排列紊乱依然存在,但未见明显大片坏死区域。结论RvD1可能通过增加肝脏HO-1表达,降低炎症因子(TNFα、IL-6、IL-8)和转氨酶(ALT、AST)水平,发挥对大鼠肝脏IR损伤的保护作用。

Objective To investigate the protective effect of resolvin D1(RvD1)in a rat model of hepatic ischemia/reperfusion(IR)injury and its association with heme oxygenase-1(HO-1).Methods A total of 36 Sprague-Dawley rats were randomly divided into sham-operation(sham)+phosphate-buffered saline(PBS)group,sham+high-dose RvD1(10μg/kg)group,IR+PBS group,IR+low-dose RvD1(2μg/kg)group,IR+middle-dose RvD1(5μg/kg)group,and IR+high-dose RvD1(10μg/kg)group,with 6 rats in each group.RvD1 were intraperitoneally injected at 1 hour before ischemia.A biochemical analyzer was used to measure the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST);ELISA was used to measure the plasma levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-8(IL-8);HE staining was used to observe the histological changes of the liver;Western blot was used to measure the change in HO-1 in liver tissue.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the IR+PBS group,the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had significant reductions in the levels of ALT,AST,and the inflammatory factors TNF-α,IL-6,and IL-8(all P<0.05),and there were no significant differences between the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group(all P>0.05).Western blot showed that compared with the IR+PBS group,the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had a significant increase in the protein expression of HO-1 in the liver(P<0.05).HE staining showed that compared with the IR+PBS group,the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had cell swelling and disordered hepatic cords,without massive necrosis.ConclusionRvD1 exerts a protective effect against hepatic IR injury in rats by increasing the expression of HO-1 and reducing the levels of inflammatory factors(TNF-α,IL-6,and IL-8)and aminotransferases(ALT and AST).