目的探讨白藜芦醇对实验性重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠肝功能的保护作用。方法96只成年雄性SD大鼠随机分为假手术(sham operation,SO)组、重症急性胰腺炎(SAP)组、地塞米松(dexamethasone,DEX)治疗组和白藜芦醇(resveratrol,RES)治疗组。各组大鼠均于制模后3、6、12h采集标本。生化方法进行肝功能测定;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定血清TNF-α和IL-6含量;透射电镜观察胰腺和肝脏超微结构的变化;RT-PCR检测凋亡调控基因Bax、Bcl-2和caspase-3 mRNA表达;Western Blot检测凋亡调控基因Bax、Bcl-2、caspase-3和线粒体内细胞色素C蛋白的表达。结果SAP组各时间点血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)和总胆红素(total bilirubin,TBIL)水平均明显高于SO组(P〈0.05),RES组和DEX组肝功损害较SAP组明显降低(P〈0.05)。RES组和DEX组TNF-α、IL-6含量均明显低于SAP组(P〈0.05)。SAP组出现明显细胞肿胀、毛细血管淤血、血栓形成、线粒体肿胀和细胞凋亡等超微结构的变化,RES组和DEX组上述改变明显减轻。RES组Bax、caspase-3 mRNA表达显著低于SAP组(P〈0.05),Bcl-2 mRNA表达明显高于SAP组。Western blot检测结果与RT-PCR结果一致。RES组各项指标与DEX组差异不显著(P〉0.05)。结论白藜芦醇可以降低SAP大鼠肝细胞凋亡的发生,从而起到减轻SAP介导的肝脏功能损伤的作用。
Objective To demonstrate the protective effect of resveratrol (RES) on the hepatic function in rats with severe acute pancreatitis (SAP). Methods Ninety-six male Sprague-Dawley (SD) rats were randomly divided into four groups: sham-operation (SO) group, SAP group, dexamethasone-treated (DEX) group and resveratroltreated (RES) group, each group having 24 rats which were evaluated at 3, 6, and 12 h during the experiment. Serum hepatic function was determined by biochemical detection. Serum TNF-α and IL-6 levels were determined by ELISA. The ultrastructure of hepatic and pancreatic tissues was observed by transmission electron microscope. The expressions of the apoptosis-related genes and proteins Bcl-2, Bax caspase-3 and cytochrome C were observed using semiquantitative RT-PCR and Western blot techniques respectively. Results The serum aspartate aminotransferase, alanine aminotransferase and total bilirubin levels were significantly higher in SAP group than in SO group. The levels of AST, ALT and TBIL in RES group were lower than those in SAP group at all the time points (P〈0.05), while there was no statistical significance between RES and DEX groups at any time points (P〉0.05). The serum TNF-α and IL-6 levels were higher in SAP group than in both RES group and DEX group (P〈0. 05). In SAP group, pancreatic and hepatic congestion, edema, inflammatory cell infiltration, mitochondrial swelling and cell apoptosis were apparent. In RES group and DEX group, pancreatic and hepatic morphological changes were alleviated at all the time points. In both RES and DEX groups, the mitochondrial membranous electric potential of hepatocytes was higher than in SAP group at any time point (P〈0.05). In comparison with those in SAP group, the expressions of Bcl-2 increased, whereas the expression of Bax, caspase-3 and cytochrome c decreased significantly in RES group (P〈0.05). The results of Western blot and RT-PCR were consistent, and no significant differences were found in al