丹芍化纤胶囊对肝纤维化大鼠肝脏Smads分子表达的影响

doi:10.3969/j.issn.1000-4718.2010.09.029

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摘要目的: 探讨丹芍化纤胶囊对大鼠实验性肝纤维化Smad2/3、Smad4和Smad7表达的影响,从而评价Smad分子作为抗纤维化靶点的应用价值。方法: 雄性Wistar大鼠48只分为正常组(n=16)、肝纤维化模型组(n=16)、丹芍化纤胶囊治疗组(n=16),除正常组外,其余采用CCl₄、饮酒、高脂低蛋白饮食等复合病因刺激制备肝纤维化动物模型8周,然后治疗组予以丹芍化纤胶囊(1 g/kg)灌胃治疗8周。实验结束后测定肝脏指数、血清透明质酸(HA)及谷丙转氨酶(ALT),光镜下观察肝组织纤维化程度,检测尿羟脯氨酸(Hyp)排出量,同时用免疫组化SABC法、免疫印迹(Western blotting)和实时荧光定量PCR (qRT-PCR) 法检测肝组织中Smad2/3、Smad4和Smad7的蛋白及mRNA的表达。结果: 治疗组与肝纤维化模型组比较,肝脏指数、血清HA及ALT显著下降,肝纤维化程度显著减轻,尿Hyp排出明显增加。治疗组肝组织中Smad7的蛋白和mRNA的表达水平与肝纤维化模型组相比均显著增高,而Smad2/3、Smad4的蛋白表达与肝纤维化模型组相比则明显降低,但mRNA水平仅轻度下降,与肝纤维化模型组比较差异不显著。结论: 中药丹芍化纤胶囊治疗大鼠肝纤维化可能主要是通过调节Smad7的基因转录和蛋白质翻译,从而发挥其反馈性抑制TGF-β/Smad促纤维化信号转导作用,这可能是丹芍化纤胶囊抗肝纤维化的作用机制之一;Smad7可能是丹芍化纤胶囊作用的一个药物靶点。

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	杨婷
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关键词 ：丹芍化纤胶囊, 四氯化碳, 肝硬化, Smad2/3, Smad4, Smad7

Abstract：AIM: To investigate the effect of Dan-shao-hua-xian(DSHX) capsule, a Chinese medicine prescription, on the expression of Smad2/3, Smad4 and Smad7, downstream signal proteins of TGF-β,in carbon tetrachloride(CCl₄)-induced rat hepatic fibrosis, and to evaluate the application of Smads as potential targets for anti-fibrosis. METHODS: Forty-eight Wistar rats were randomly divided into three groups: normal control group, CCl₄-induced hepatic fibrosis group and DSHX treatment group. The rats in both CCl₄-induced hepatic fibrosis group and DSHX treatment group were induced by subcutaneous injection of CCl₄, drinking alcohol, giving diet of hyperliprosis and hypoprotein for 8 weeks to establish the hepasis fibrosis model. The hepatic fibrosis rats in DSHX treatment group were treated with DSHX capsule (1 g/kg, po) once per day for 8 weeks. Finally, the liver index, serum hyaluronic acid (HA), alanine aminotransferase (ALT), the degree of hepatic fibrosis and the urinary excretion of hydroxyproline (Hyp) were evaluated. Meanwhile, the techniques of immunohistochemistry, Western blotting and qRT-PCR were applied to detect the expression of Smad2/3 and Smad4 and Smad7 at both mRNA and protein levels. RESULTS: Compared to hepatic fibrosis group, the liver index, serum levels of HA and ALT, and the degree of hepatic fibrosis were all significantly reduced, and the urinary excretion of Hyp was increased in DSHX treatment group. The mRNA and protein expression of Smad7 in the liver tissues in DSHX treatment group significantly increased as compared to that in hepatic fibrosis group. However, the protein expression of Smad2/3 and Smad4 decreased and the mRNA expression of Smad2/3 and Smad4 was unchanged as compared to that in hepatic fibrosis group. CONCLUSION: Dan-shao-hua-xian capsule promotes the transcription and expression of Smad7 to exert its feedback suppression on TGF-β/Smad-induced hepatic fibrosis signal transduction pathway. This might be one of the mechanisms by which Dan-shao-hua-xian capsule inhibits the process of hepatic fibrosis, and Smad7 might be the potential target.

Key words： Dan-shao-hua-xian capsule Carbon tetrachloride Liver cirrhosis Smad2/3 Smad4 Smad7

收稿日期: 2010-01-05

中图分类号:

R363.2

通讯作者: 杨勤

引用本文:

杨婷; 谢汝佳; 罗新华; 杨勤. 丹芍化纤胶囊对肝纤维化大鼠肝脏Smads分子表达的影响[J]. 中国病理生理杂志, 2010, 26(9): 1807-1812. YANG Ting, XIE Ru-jia, LUO Xin-Hua, YANG Qin. Effects of traditional Chinese medicine prescription Dan-shao-hua-xian capsule on expression of Smads in rat liver of hepatic fibrosis. Chin J Pathophysiol, 2010, 26(9): 1807-1812.

链接本文:

http://www.cjpp.net/CN/10.3969/j.issn.1000-4718.2010.09.029 或 http://www.cjpp.net/CN/Y2010/V26/I9/1807

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