柴胡皂苷d对免疫性肝纤维化大鼠TGF-β1、HYP、SOD、MDA的影响

目的构建大鼠免疫性肝纤维化模型,探讨柴胡皂苷D(SS-d)抗模型鼠肝纤维化的药理学活性及其相关机制。方法猪血清免疫法建立大鼠肝纤维化模型,实验分为6组:正常对照组、模型组、高中低剂量SS-d用药组(20、10、5mg/kg)及秋水仙素(Col,0.1mg/kg)用药组。放射免疫分析法测定大鼠血清透明质酸酶(HA)、层粘连蛋白(LN)和III型前胶原(PCIII)的水平。分光分析法检测标本中超氧化物歧化酶(SOD)活性和羟脯氨酸(Hyp)、丙二醛(MDA)的含量。荧光定量PCR和Westernblot检测肝组织中TGF-β1mRNA和蛋白表达的变化。结果成功构建了免疫性大鼠肝纤维化模型,模型鼠中血清HA、LN和PCIII显著高于正常对照组(P﹤0.01),SS-d可以剂量依赖性地抑制模型组血清中HA、LN和PCIII肝纤维化指标的升高(P﹤0.05)。与模型组比较SS-d可以剂量依赖性地减低肝组织中Hyp和MDA的含量,同时也剂量依赖性地升高SOD的活性(P﹤0.05)。荧光定量PCR和Westernblot检测结果显示SS-d可以剂量依赖性地下调模型鼠肝组织中TGF-β1mRNA和蛋白的表达水平。结论 SS-d具有较好的抗肝纤维化作用,其作用机制可能与提高SOD活性、减少氧自由基的生成、抑制TGF-β1信号通路的活化有关。

Effect of Saikosaponin D on immunity hepatic fibrosis in Rats

OBJECTIVE To establishment the rat model of liver fibrosis by porcine serum, and to study the effect of Saikosaponin D (SS-d) on liver fibrosis indexes. METHODS The model of immunity hepatic fibrosis was established by porcine serum. The rats received different treatment and divided into six groups, including control group, model group, high-dose SS-d group, medium-dose SS-d group, low-dose SS-d group and colchicine (Col) group. Col was used as a passive control. The serum level of HA, LN and PCIII was determined by radioimmunity assay. The activity of SOD and contents of MDA and Hyp was detected by spectrophotometer. Real-time PCR and Western blot was used to detect the mRNA and protein expression of TGF-β1, respectively. RESULTS The serum level of HA, LN and PCIII was higher in model rat than those of controls (P﹤ 0.01). The content of HA, LN, PCIII, MDA and Hyp was decreased by SS-d treatment in a dose-dependent manner (P﹤ 0.05). The activity of SOD in model rat was also increased by SS-d treatment (P﹤0.05). The mRNA and protein expression of TGF-β1 was decreased by Real-time PCR and Western blot analysis, respectively. CONCLUSION SS-d showed a potent activity in anti-liver fibrosis. The upregulation of SOD activity and downregulation of TGF-β1expression might contribute to the action mechanisms of SS-d against liver fibrosis.