柴胡疏肝散对肝纤维化大鼠TGF-β_1/Smad信号通路的影响

Effects and Mechanism of Chaihu Shugan Powder on TGF-β_1/Smad Signaling Pathways in Hepatic Fibrosis Model Rats

目的:探讨柴胡疏肝散对肝纤维化(hepatic fibrosis,HF)的作用机制。方法:60只Wistar大鼠被随机均分为6组:正常组、模型组、柴胡疏肝散高、中、低剂量组、柴胡疏肝散预防组。除正常组外,其余各组均采用猪血清ip诱发HF,0.5 m L/只,2次/周,连续10周,5周后即可形成HF。预防组于造模同时给药(以柴胡疏肝散6.3 g·kg-1),柴胡疏肝散高、中、低剂量组[(12.6,6.3,3.15)g·kg-1]于造模第6周给药,连续10周。采用RT-PCR法检测各组肝组织转化生长因子-β1(TGF-β1),Smad2,Smad3,Smad4和Smad7的mRNA表达,免疫组化法分别检测肝组织TGF-β1,磷酸化Smad 2/3(p-Smad 2/3)和Smad7蛋白表达。结果:与正常组比较,模型组TGF-β1,Smad2,Smad3的mRNA和TGF-β1,p-Smad 2/3蛋白表达均显著增加(均P<0.01),Smad7基因表达显著降低(均P<0.01);与模型组比较,柴胡疏肝散中剂量组TGF-β1和Smad3 mRNA表达均显著降低,p-Smad 2/3和TGF-β1蛋白表达显著减弱(P<0.05,P<0.01),Smad7基因表达显著增加(P<0.01)。结论:柴胡疏肝散有明显的抗HF作用,其机制可能与抑制TGF-β1/Smad信号通路有关。

Objective： To investigate the protective effects and mechanism of Chaihu Shugan powder （CHSGS） on hepatic fibrosis in rats. Method： Sixty Wistar rats were randomly divided into 6 groups： normol group, model group, Chaihu Shugan powder high-dose, mediate-dose, low-dose, and prevention group, except the normol group, all the other groups were injected with pig＇s serum without inactivation into abdominal cavity, twice per week and 0.5 mL each time, persisting for 10 weeks. The prevention groups were injected the stomach with the medicines （Chaihu Shugan powder 6.3 g·kg^-1, 10 weeks） at the same time when models began to be made, the other treatment groups were dealed with just as the prevention groups 6 weeks later persisting for 4 weeks. Then rats were randomly taken for decollation in all 6 groups at the end of 10th week. Chaihu Shugan powder high-dose group 12.6 g·kg^-1 , mediate-dose group 6.3 g·kg^-1 , low-dose group 3. 15 g·kg^-1. RT-PCR assay were used to detect the expression of transforming growth factor-fl~ （TGF-βl）, Smad2, Smad3, Smad4 and Smad7mRNA. The expression of TGF-β1, p-Smad 2/3 and Smad7 protein in hepatic fibrosis tissues was detected by immunohistochemical techniques. Result： The expression of TGF-βl, Smad2, Smad3 mRNA and the expression of TGF-β1, p-Smad 2/3 protein were markedly higher, while the expression of Smad7 gene were significantly lower in the model group than those in normal group （P 〈 0.01 ）. Compared with the model group, the expression of TGF-β1 and Smad3 mRNA were markedly lower, but the expression of Smad7 gene were significantly higher, the expression of p-Smad 2/3 and TGF-β1, protein were markedly lower （P 〈 0.05, P 〈 0.01 ）in Chaihu Shugan powder mediate-dose group. Conclusion： Chaihu Shugan powder have a positive action on hepatic fibrosis model rats. The mechanism of its effect is related with its inhibiting the expression of TGF-β1, Smad3 genes, and p-Smad 2/3 protein, and enhancing the expression of Smad7 gene in rats liver.