白藜芦醇对梗阻性黄疸大鼠肝损害的保护机制

Protective mechanism of the resveratrol in liver injury of obstructive jaundice in rats

目的 探讨白藜芦醇（Res）对大鼠梗阻性黄疸肝损害的保护作用及机制.方法 将雄性Wistar大鼠60只随机分为3组,A组：假手术组,B组：胆道结扎组,C组：胆道结扎同时Res干预组.取血清检测总胆红素（TB）、直接胆红素（DB）、谷丙转氨酶（A LT）,Western blot检测肝组织沉默信息调控因子1（SIRT1）、核因子-κB （NF-κBp65）蛋白表达,实时定量聚合酶链反应（Real-timePCR）测肝组织SIRT1 mRNA及过氧化体增殖剂激活受体α（PPARα） mRNA,免疫组织化学法测肝组织PPARα蛋白表达,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）法测肝细胞凋亡.结果 B组与A组比较：血清ALT升高、SIRT1 mRNA及蛋白和PPARα mRNA及蛋白降低NF-κBp65蛋白升高、细胞凋亡率升高（P＜0.05）;C组与B组比较：血清ALT降低、SIRT1 mRNA及蛋白和PPARα mRNA及蛋白增加、NF-κBp65蛋白降低、细胞凋亡率降低（P＜0.05）;A、B、C组ALT、SIRT1 mRNA及蛋白、PPARαmRNA及蛋白、NF-κBp65、凋亡率指标为：A组（42.54±4.36） IU/L、1.000±0.000、0.320±0.007、1.000±0.000、73 134.33±6 775.63、0.14±0.02、（0.28±0.08）％;B组：（240.53 ±12.04） IU/L、0.430±0.030、0.190±0.005、0.390 ±0.028、38 417.33±3 039.34、0.54±0.05、（12.78±1.50）％;C组：（157.13 ±8.01） IU/L、0.740±0.027、0.240±0.012、0.630±0.031、55 173.00±4 222.83、0.41 ±0.04、（6.15±1.06）％.结论 Res能减轻梗阻性黄疸肝损害,可能通过激活SIRT1抑制NF-κB发挥抗炎、抗凋亡作用,促进PPARα的表达而发挥抗氧化作用.

Objective To explore the protective mechanism and effect of the resveratrol （Res） for liver injury of obstructive jaundice.Methods The rats were divided randomly into three groups：the sham group receiving laparotomy without bile duct ligation （BDL）,the BDL group and the BDL ＋ Res group with the Res given following BDL.The expression of the silent information regulator 1 （SIRT1） and nuclear factor-κBp56 （NF-κB） proteins were analyzed by western blotting but immunocytochemical assay was performed to examine peroxisome proliferator activated receptor-alpha （PPARα） protein.Real-time polymerase chain reaction （PCR） was performed to determine the mRNA expression of SIRT1 and PPARα.Cell apoptosis was examined by terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL） staining.Results After the BDL,the cholestasis model was established.Compared B group to A group,the alanine transaminase （ALT） was higher and the mRNA and proteins expression of the SIRT1 and the PPARα was lower,but the NF-κB protein and the rate of cell apoptosis was higher （P 〈 0.05）.Compared B group to C group,the ALT was higher and the mRNA and proteins expression of the SIRT1 and the PPARα was lower,but the NF-κB protein and the rate of cell apoptosis was significantly higher （P 〈 0.05）.Conclusion This study demonstrates that the Res could alleviate liver damage and that the Res plays a beneficial role to resist inflammation and apoptosis by activating the SIRT1 which probably inhibits the expression of NF-κB protein in cholestatic liver injury.The Res also shows antioxidant effect by promoting the expression of PPARα .