温阳化浊通络方对系统性硬皮病成纤维细胞周期和增殖的影响

Effect of Wenyang Huazhuo Tongluo Recipe Contained Serum on Proliferation and Cell Cycle of Systemic Sclerosis Skin Fibroblasts

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)皮肤成纤维细胞周期和增殖的影响。方法:通过SSc患者制备强的松、青霉胺及温阳化浊通络方含药血清,分别加入皮肤成纤维细胞培养体系中。培养48h后,四甲基偶氮唑蓝(MTT)法和流式细胞仪分别测定细胞增殖和细胞周期。结果:与对照组相比,不同浓度的西药组、中药组、中西医结合组含药血清均能明显抑制正常人和SSc皮损成纤维细胞的增殖(P<0.05或P<0.01)。与西药组相比,20%中西医结合组血清抑制作用更加明显(P<0.01)。不同浓度的中药组血清能显著提高SSc皮损成纤维细胞的G0-G1期细胞比例(P<0.01),降低S期细胞和G2-M期细胞比例(P<0.05或P<0.01)。结论:温阳化浊通络方通过抑制SSc皮损成纤维细胞进入细胞分裂周期,对其增殖具有明显的抑制作用。同时,温阳化浊通络方对体外培养的正常皮肤和SSc皮损成纤维细胞的干预作用不具有选择性。

Objective： To investigate the effect of ＂Wenyang Huazhuo Tongluo recipe＂ （WYHZTLR） on proliferation and cell cycle of systemic sclerosis （SSc） skin fibroblasts in vitro by serum pharmacologic technique. Methods ： Prednisone, D-penicillamine and WYHZTLR contained serum from patients with SSc were respectively added into cultured normal dermal fibroblast cell-line and SSc le- sional dermal fibroblast cell-line. Then the cells were incubated for 48 hours. Cell proliferation and cell cycle were examined by methyl thiazolyl tetrazolimn （MTT） assay and flow cytometry, respectively. Results： Western medicine Chinese medicine and integrated Chi- nese and western medicine could markedly inhibit the proliferation of normal dermal fibroblasts and SSc lesional dermal fibroblasts in vitro compared with control group （P 〈0. 05 or P 〈0. 01 ）. But 20% serum in integrated Chinese and western medicine group could markedly inhibit the proliferation of fibroblasts compared with western medicine group （ P 〈 0.01 ）. Different drug-contained serum of WYHZTI,R could markedly increase the percentage of G0-G1 stage cells （ P 〈 0. 01 ） and depress the percentage of sphase ceils and G2-M stage cells compared with control group （P 〈0. 05 or P 〈0. 01 ）. Conclusion：WYHZTLR can block the progression of SSc skin fibroblasts into sphase and G2-M stage and inhibit the proliferation of SSc skin fibroblasts, and WYHZTLR was unselectively interfered with normal and SSc fibrohlasts in vitro.