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目的:运用血清药理学方法,探讨温阳化浊通络方对系统性硬皮病(SSc)皮肤成纤维细胞胶原分泌和转化生长因子β1(TGF-β1)的影响。方法:对SSc患者分别采用强的松(每日20 mg)和青霉胺(每日0.25 g)(西药组),温阳化浊通络方每日1剂,分早晚2次服用(中药组),西药加中药(中西药结合组)治疗1个月后,制备含药血清,加入皮肤成纤维细胞培养体系中。培养72 h后,采用比色法和ELISA法分别测定成纤维细胞培养基中羟脯氨酸(Hyp)和TGF-β1含量。结果:与对照组相比,浓度为5%,10%,20%的西药组、中药组、中西药结合组含药血清均能明显抑制正常人和SSc皮肤成纤维细胞胶原分泌(P<0.05或P<0.01);与西药组相比,中西药结合组抑制作用更加明显(P<0.01)。浓度为5%,10%,20%的西药组、中药组、中西药结合组含药血清能显著抑制SSc皮肤成纤维细胞TGF-β1的表达(P<0.01),且中西药结合组比西药组的抑制作用更强(P<0.01)。同时,对照组SSc皮肤成纤维细胞培养基中胶原含量与TGF-β1呈直线正相关性(r=0.895,P<0.05)。结论:温阳化浊通络方能有效减少体外培养的SSc皮肤成纤维细胞胶原和TGF-β1的分泌,可能是其治疗SSc的药理作用机制之一。
Objective: To explore the effect of Wenyang Huazhuo Tongluo Prescription on collagen secretion and transforming growth factor-β1 (TGF-β1) in the skin fibrosis cells of patients with systemic scleroderma (SSc) by using serum pharmacology method. Methods: Patients with SSc were treated with prednisone (20 mg daily) and penicillamine (0.25 g daily) (western medicine group), and once daily with Wenyang Huazhuo Tongluo Recipe Group), Western medicine plus traditional Chinese medicine (Chinese and Western medicine group) for 1 month after treatment, preparation of serum containing, added to the skin fibroblast culture system. After cultured for 72 h, the content of hydroxyproline (Hyp) and TGF-β1 in fibroblast culture medium were measured by colorimetric method and ELISA method respectively. Results: Compared with the control group, the drug-containing serum of the western medicine group, the traditional Chinese medicine group and the traditional Chinese medicine combined with western medicine group could inhibit the collagen secretion of normal human and SSc dermal fibroblasts at 5%, 10% and 20%, respectively (P <0.05 Or P <0.01). Compared with Western medicine group, the inhibitory effect of Chinese medicine and western medicine combination group was more obvious (P <0.01). Western medicine group, traditional Chinese medicine group and traditional Chinese medicine combined group could significantly inhibit the expression of TGF-β1 in SSc skin fibroblasts (P <0.01) with the concentrations of 5%, 10% and 20% The inhibition of the group was stronger (P <0.01). At the same time, collagen content in the culture medium of SSc skin fibroblasts was positively correlated with TGF-β1 (r = 0.895, P <0.05). Conclusion: Wenyang Huazhuo Tongluo decoction can effectively reduce the secretion of collagen and TGF-β1 in SSc skin fibroblasts cultured in vitro, which may be one of the pharmacological mechanisms of its treatment of SSc.