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目的:观察针刺结合温阳化浊通络方对系统性硬化病(SSc)大鼠模型皮肤成纤维细胞转化生长因子(TGF)信号通路的影响,揭示其抗纤维化作用机制。方法:将无特定病原体(SPF)级大鼠随机分为5组:空白对照组、模型对照组、温阳化浊通络方组、针刺组、针刺加温阳化浊通络方组,造模成功后,每天针刺一次,温阳化浊通络方按10ml/kg容量灌胃,连续4周。检测皮肤组织成纤维细胞TGF-β1、羟脯氨酸(Hyp)、结缔组织生长因子(CTGF)、CTGF mRNA表达水平及TGF-β1对CTGF蛋白表达的影响。结果:经不同方法治疗后,组内比较CTGF、TGF-β1、Hyp的含量,有显著差异(P<0.01)。CTGF mRNA、CTGF蛋白表达相比,有显著差异(P<0.05,P<0.01)。最小显著差法(LSD)两两相比,除温阳化浊通络方组和针刺组两组之间(P>0.05)无显著差异外,其他各组之间TGF-β1、Hyp、CTGF、CTGF mRNA表达水平相比(P<0.01),有显著差异。TGF-β1可以显著上调CTGF的蛋白表达(P<0.01),使用温阳化浊通络方可以下调CTGF(P<0.01),与TGF-β1联合使用时,可以抑制TGF-β1对CTGF的上调作用。结论:针刺结合温阳化浊通络方可以在mRNA水平和蛋白水平起作用,从而调节TGF信号通路,这可能是针刺结合温阳化浊通络方抑制SSc的重要机制。
Objective: To observe the effect of acupuncture combined with Wenyang Huazhuo Tongluo Recipe on TGF-β signaling pathway in the skin fibroblast of the rat model of systemic sclerosis (SSc), and to reveal its anti-fibrosis mechanism. Methods: SPF rats were randomly divided into 5 groups: blank control group, model control group, Wenyang Huazhuo Tongluo Prescription group, acupuncture group, acupuncture plus Wenyang Huazhuo Tongluo Prescription group , After the success of modeling, acupuncture once a day, Wen Yang Huazhuo Tongluo side by 10ml / kg capacity of gavage for 4 weeks. The expression of TGF-β1, Hyp, CTGF and CTGF mRNA and the expression of CTGF in TGF-β1 were detected. Results: After treatment by different methods, the contents of CTGF, TGF-β1 and Hyp in the group were significantly different (P <0.01). CTGF mRNA and CTGF protein expression were significantly different (P <0.05, P <0.01). There was no significant difference between the two groups (P> 0.05) except for Wenyang Huazhuo Tongluo Formula group and the acupuncture group (P <0.05). The levels of TGF-β1, Hyp, CTGF, CTGF mRNA expression levels compared (P <0.01), a significant difference. TGF-β1 significantly upregulated the expression of CTGF protein (P <0.01). The treatment of Wenyang Huazhuo Tongluo Decoction could down-regulate CTGF (P <0.01). When combined with TGF-β1, TGF-β1 inhibited the up-regulation of CTGF effect. Conclusion: Acupuncture combined with Wenyang Huazhuo Tongluo Recipe can play a role in mRNA and protein levels and thus regulate TGF signaling pathway, which may be an important mechanism of acupuncture combined with Wenyang Huazhuo Tongluo Fang in inhibiting SSc.