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目的探讨温阳化浊通络方治疗系统性硬化症(SSc)的可能作用机制。方法将60只C3H/He小鼠随机分为正常组、模型组、西药组和中药低、中、高剂量组,每组10只。除正常组外其余各组给予皮下注射博莱霉素3周建立SSc小鼠模型。西药组小鼠灌服青霉胺片65 mg/(kg·d),中药低、中、高剂量组分别灌服温阳化浊通络方水煎剂11.75、23.50、47.00 g/(kg·d),模型组和正常组给予等体积的生理盐水,共干预8周。采用HE染色观察皮肤组织病理变化,采用RT-PCR法检测皮肤转化生长因子β1(TGF-β1)、Smad2、Smad7 mRNA表达,采用Western blot法检测皮肤Ⅰ、Ⅲ型胶原蛋白(COL-Ⅰ、COL-Ⅲ)表达。结果模型组较正常组皮肤真皮变厚,胶原增加、变粗,真皮小血管周围炎症细胞浸润,西药组和中药低、中、高各剂量组与模型组比较真皮变薄、胶原含量减少,血管周围炎症细胞浸润减轻。与正常组比较,模型组TGF-β1、Smad2 mRNA及COL-Ⅰ、COL-Ⅲ蛋白表达升高,Smad7 mRNA表达降低(P<0.01);与模型组比较,西药组和中药低、中、高剂量组TGF-β1、Smad2 mRNA及COL-Ⅰ、COL-Ⅲ蛋白表达减少,Smad7 mRNA表达增加(P<0.05或P<0.01);中药高剂量组TGF-β1、Smad2、Smad7 mRNA及COL-Ⅰ、COL-Ⅲ蛋白表达改善优于西药组和中药低剂量组(P<0.05或P<0.01)。结论温阳化浊通络方可拮抗SSc纤维化,可能与干预TGF-β1/Smad信号通路、降低皮肤胶原蛋白含量有关,且高剂量效果更佳。
Objective To investigate the possible mechanism of Wenyang Huazhuo Tongluo Recipe in treating systemic sclerosis (SSc). Methods Sixty C3H / He mice were randomly divided into normal group, model group, western medicine group and traditional Chinese medicine low, medium and high dose group, 10 rats in each group. In addition to the normal group the remaining groups were given subcutaneous injection of bleomycin 3 weeks SSc mouse model. Western medicine group mice were fed penicillamine tablets 65 mg / (kg · d), Chinese medicine low, medium and high dose groups were fed Wenyang Huazhuo Tongluo Decoction 11.75,23.50,47.00 g / (kg · d), model group and normal group were given equal volume of saline, a total of 8 weeks intervention. The pathological changes of the skin tissues were observed by HE staining. The expressions of TGF-β1, Smad2 and Smad7 mRNA were detected by RT-PCR. The expressions of collagen type Ⅰ and Ⅲ (COL-Ⅰ, COL -III) expression. Results Compared with the model group, the dermis in the model group was thickened, the collagen was increased and thicker, the infiltration of inflammatory cells in the dermis and perivascular area was lower. Compared with the model group, the dermal thinning, collagen content, Peripheral inflammatory cell infiltration reduced. Compared with the normal group, the expression of TGF-β1, Smad2 mRNA and COL-Ⅰ and COL-Ⅲ protein in the model group were increased and the expression of Smad7 mRNA was decreased (P <0.01). Compared with the model group, The expression of TGF-β1, Smad2 mRNA, COL-Ⅰ and COL-Ⅲ protein and the expression of Smad7 mRNA in the dose group were significantly increased (P <0.05 or P <0.01) , COL-Ⅲ protein expression improved better than Western medicine group and traditional Chinese medicine low-dose group (P <0.05 or P <0.01). Conclusion Wenyang Huazhuo Tongluo can antagonize SSc fibrosis, which may be related to the intervention of TGF-β1 / Smad signaling pathway and the reduction of skin collagen content, and the high-dose effect is better.