三七总甙对人增生性瘢痕成纤维细胞转分化的作用

目的 观察三七总甙（PNS）对人增生性瘢痕成纤维细胞（HSF）向肌成纤维细胞转分化的作用，探讨PNS抗瘢痕纤维化的机制。方法 体外培养HSF，采用不同浓度的PNS进行干预，根据细胞培养所加PNS浓度分为400μg／ml组、800μg／ml组及空白对照组（不加PNS）。采用细胞三维培养法检测HSF凝胶收缩情况，计算其收缩指数；免疫细胞化学染色法检测HSF中“平滑肌肌动蛋白（α-SMA）的表达；流式细胞仪检测HSF中α-SMA阳性细胞率。结果 400μg／ml、800μg／ml组各时相点的胶原凝胶块收缩程度明显减轻，其收缩指数均小于空白对照组（P〈0．05或P〈0．01）。HSF中α-SMA阳性表达颗粒在细胞质内呈弥漫性分布；空白对照组HSF中α-SMA的阳性表达明显强于400μg／ml、800μg／ml组。400μg／ml、800μg／ml组α-SMA的阳性细胞率（31．52％、24．28％）均明显低于空白对照组（45．74％，P〈0．05）。400l,Lg／ml、800μg／ml组α-SMA的阳性细胞染色强度积分均明显低于空白对照组（P〈0．05或P〈0．01）。结论 PNS能够抑制HSF向肌成纤维细胞的转分化，具有体外抗瘢痕纤维化的作用。

Efffects of Panax notogin seng on the transdifferentiation of fibroblasts in human hypertrophic scar in vitro

OBJECTIVE: To observe the effects of Panax notogin seng on the transdifferentiation of the cultured human fibroblasts from hypertrophic scar in vitro, and explore its anti-fibrosis mechanism. METHODS: The fibroblasts from human hypertrophic scar were cultured in vitro. Different amount of Panax notogin seng was added into the medium, respectively (400 microg/ml and 800 microg/ml). A culture without addition of the drug served as control. The fibroblast-populated collagen lattice method was used to detect the gel contraction, and contraction ratio was calculated. The immunocytochemistry staining method was used to detect the expression of alpha-smooth muscle actin. The flow cytometry method was used to detect the positive rate of alpha-smooth muscle actin. RESULTS: The contraction degree of the fibroblasts after PNS administration was ameliorated at each time-point, with contraction index lower than that of controls (P < 0.05 or P < 0.01). Scattered distribution of alpha-SMA positive granules were observed in the cytoplasma, and the positive rate of alpha-SMA expression in 400 microg/ml (31.52%) and 800 microg/ml (24.28%) PNS groups were obviously lower than that in control group (45.74%, P < 0.05). The staining intensity of positive cells in 400 microg/ml and 800 microg/ml PNS groups was also obviously lower than that in control group (P < 0.05 or P < 0.01). CONCLUSION: Panax notogin seng can inhibit the transdifferentiation of the cultured human fibroblasts from hypertrophic scar, and it exhibits an anti-fibrosis effect on hypertrophic scar in vitro.