白藜芦醇对胆道梗阻再通大鼠肝功能的保护作用

Protective effect of resveratrol on liver injury in rats after recanalization of biliary obstruction

目的研究白藜芦醇(Res)对胆道梗阻再通大鼠肝损害的保护作用及机制。方法雄性Wistar大鼠60只,随机分为4组,即A组:假手术组,B组:梗阻性黄疸模型组,C组:梗阻性黄疸模型+胆道再通组,D组:梗阻性黄疸+胆道再通+Res干预组。连续给药7 d,实验结束后检测各组大鼠血清中总胆红素(TBIL)、直接胆红素(DBIL)、丙氨酸氨基转氨酶(ALT)水平;RT-PCR检测肝组织沉默信息调控因子1(SIRT1)m RNA的表达,Western blot检测肝组织SIRT1蛋白和核因子-κB(NF-κB)蛋白表达,免疫组化检测过氧化物酶体增殖物激活受体α(PPARα)蛋白在肝脏中的表达,原位末端标记(TUNEL)法检测肝细胞凋亡。结果与假手术组比较,模型组血清ALT水平升高,SIRT1 m RNA及蛋白、PPARα蛋白表达降低,NF-κB蛋白表达升高,细胞凋亡率升高(P<0.05);而造模后胆道再通组与模型组比较,血清ALT水平降低,SIRT1 m RNA及蛋白、PPARα蛋白表达增加,NF-κBp蛋白表达降低,细胞凋亡率降低(P<0.05);白藜芦醇组与C组比较:血清ALT水平降低,SIRT1 m RNA及蛋白、PPARα蛋白表达增加,NF-κBp蛋白表达降低,细胞凋亡率降低(P<0.05)。结论 Res可能通过激活SIRT1抑制NF-κB,发挥抗炎、抗凋亡作用,通过促进PPARα的表达发挥抗氧化作用,从而减轻胆道梗阻再通大鼠的肝损害,促进肝功能恢复。

Objective To explore the protective effect of resveratrol （Res）on liver injury in rats after recanalization of biliary obstruc-tion.Methods Sixty healthy male Wistar rats were randomized into four groups：sham group （group A）,obstructive jaundice one week group （group B）,obstructive jaundice one week and recanalization one week ＋NS group （group C）,obstructive jaundice one week and recanalization one week ＋Res group （group D）.The levels of total bilirubin（TBIL）,direct bilirubin（DBIL）and serum ala-nine aminotransferase（ALT）were measured.Real-time polymerase chain reaction （RT-PCR）was performed to determine the mRNA expression of silent information regulator 1（SIRT1）.The expression of the SIRT1 and nuclear factor-κB （NF-κB）proteins were detec-ted by Western blot.Immunocytochemical assay was performed to examine peroxisome proliferator activated receptor-alpha （PPARα） protein.Hepatocellular apoptosis was examined by terminal deoxynucleotidyl transferase mediated nick end labeling （TUNEL）method. Results Compared with group A,in group B the level of ALT was higher,the expressions of SIRT1 mRNA and protein and the PPARαprotein were lower,and the NF-κB protein and the rate of hepatocellular apoptosis were higher（P 〈0.05）.Compared with group B,in group C the level of ALT was lower,the expression of SIRT1 mRNA and protein and the PPARαprotein were higher,and the NF-κB protein and the rate of hepatocellular apoptosis were lower（P 〈0.05）.Compared with group C,in group D the level of ALT was lower,the expressions of SIRT1 mRNA and protein and the PPARαprotein were higher,and the NF-κB protein and the rate of hepatocellular apoptosis were lower（P 〈0.05）.Conclusion The Res could resist inflammation and apoptosis by activating the SIRT1 which probably inhibits the expression of NF-κB protein,and palys an antioxidant role by promoting the expression of PPARαin rats after recanalization of biliary obstruction,so that it could alleviate liver damage.