雄芍汤调节TGF-β/Smad通路有效部位的研究

摘要

This study was aimed to study the therapeutic material basis of Xiong-Shao decoction (XSD) on hepatic fi-brosis (HF), and to screen the effective parts from XSD for regulating TGF-β/Smad pathway. Male Wistar rats were randomly divided into the normal group, the model group, the Fu-Zheng Hua-Y u (FZHY) capsule group, the XSD group, the crude polysaccharide group, the total glycosides group, and the total alkaloids group. Rats of the modeling group were intraperitoneally injected with dimethylnitrosamine (DMN) to establish HF model. After modeling, FZHY capsule solution (0.105 g·mL-1), XSD crude polysaccharides extract solution (35.420 mg·mL-1), XSD total glycosides extract solution (25.725 mg·mL-1), and XSD total alkaloids extract solution (0.196 mg·mL-1) were administered to the corresponding treatment group by gavage once a day for 4 weeks, respectively. Rats of the normal group and the model group were given equivalent amount of normal saline by gavage once a day for 4 weeks. The treatment course was 4 weeks. The expression of TGF-β1 mRNA of the liver tissues was detected by FQ-PCR. And the protein ex-pressions of Smad3 and Smad7 were detected by western blotting analysis. The results showed that compared with the model group, there was no significant difference in the expression of Smad7 protein between the total glyco-sides group and the model group, as well as no significant difference between the total alkaloids group and the model group. Expressions of TGF-β1 mRNA and Smad3 protein of other treatment groups were significantly re-duced. And the expressions of their Smad7 protein were significantly increased (P < 0.05 or P < 0.01). It was concluded that crude polysaccharide an d total glycosides fractions were the effective parts of XSD for regulating TGF-β/Smad pathway.%目的：研究雄芍汤抗肝纤维化的药效物质基础，筛选其调节TGF-β/Smad通路的有效部位。方法：Wistar雄性大鼠随机分为正常组、模型组、扶正化瘀胶囊组（扶正组）、雄芍汤组（雄芍组）、粗多糖组（多糖组）、总苷组、总生物碱组（总碱组），二甲基亚硝胺（DMN）腹腔注射复制大鼠肝纤维化模型。造模成功后，开始灌胃给药，治疗组分别给予扶正化瘀胶囊（0.105 g·mL-1）、雄芍汤粗多糖提取物（35.420 mg·mL-1）、雄芍汤总苷提取物（25.725 mg·mL-1）、雄芍汤总生物碱提取物（0.196 mg·mL-1），正常组与模型组灌服等量生理盐水，每日1次，疗程4周。4周后处死取材，实时荧光定量PCR检测肝组织TGF-β1 mRNA表达， Western blotting检测肝组织Smad3、Smad7蛋白表达。结果：与模型组比较，各治疗组TGF-β1 mRNA、Smad3蛋白表达均显著降低，Smad7蛋白表达均显著升高（P<0.05或P<0.01），总苷组Smad7、总碱组与模型组无显著性差异。结论：粗多糖部位和总苷部位为雄芍汤调节TGF-β/Smad通路的有效部位。