转化生长因子β1和胰岛素样生长因子Ⅰ及其协同作用对人皮肤成纤维细胞增殖的影响

Effect of transforming growth factor beta 1, insulin-like growth factor-Ⅰ and their synergy on the proliferation of human skin fibroblasts

背景:皮肤成纤维细胞注射移植为颜面部皱纹及微小凹陷等治疗提供了一种全新的方法,然而成纤维细胞的获取及体外快速扩增往往需要较长时间,以至于不能满足临床需要。目的:观察转化生长因子β1、胰岛素样生长因子Ⅰ及两者协同作用对体外培养人皮肤成纤维细胞增殖的影响。方法:人皮肤成纤维细胞原代培养,取第3代细胞种入96孔板,加入不同质量浓度的转化生长因子β1或胰岛素样生长因子Ⅰ作用于细胞,分别作用3,6,9d采用MTT法检测细胞增殖情况。同法设4个组,分别为:阴性对照组、转化生长因子β1最佳效应浓度组、胰岛素样生长因子Ⅰ最佳效应浓度组、两者最佳效应浓度联用组,作用3,6,9d采用MTT法检测增殖情况。结果与结论:作用6d和9d,10.0μg/L转化生长因子β1、50.0μg/L胰岛素样生长因子Ⅰ组与对照组相比均有促细胞增殖效应(P<0.05),此即为各生长因子最佳效应浓度。作用6d和9d,转化生长因子β1与胰岛素样生长因子Ⅰ各浓度实验组与相应对照组相比,差异均有显著性意义(P<0.05)。10.0μg/L转化生长因子β1与50.0μg/L胰岛素样生长因子Ⅰ联合作用9d可显著促进细胞增殖,与其他组相比,差异有显著性意义(P<0.05)。提示转化生长因子β1、胰岛素样生长因子Ⅰ均可体外促进人皮肤成纤维细胞增殖,最佳效应浓度联用促增殖效果优于各自单独使用。

BACKGROUND： The application of injecting human skin fibroblast （HSF） cultured in vitro has provided a firenew idea for the treatment of facial wrinkles and minor depression. However, it would be more practical if the time which HSF cultured in vitro could be shortened. OBJECTIVE： To investigate the effect of transforming growth factor-β1 （TGF-β1）, insulin-like growth factor-I （IGF-Ⅰ） and their synergy on the HSF proliferation. METHODS： HSF were cultured in vitro. The 3rd passage cells were inoculated into 96-well tissue culture plates, and treated with different concentrations of TGF-β1, IGF-Ⅰ, respectively, and the combinations of TGF-β1 and IGF-Ⅰwere established at their optimal effect concentrations. The control group was also established for comparison. The proliferation was detected at 3, 6 and 9 days by the MTT colorimetric method. RESULTS AND CONCLUSION： The proliferating effects of two growth factors （10.0 μg/L TGF-β1, 50.0 μg/L IGF-Ⅰ） on the 3rd passage cells at 6 and 9 days were significantly better than in the control group （P〈0.05）, thus, this was the optimal concentrations for growth factors. The differences between the experimental group and corresponding control groups had significance at 6 and 9 days after operation （P〈0.05）. Compared with other groups, the combination groups of 10.0 μg/L TGF-β1 and 50.0 μg/L IGF-Ⅰ showed a significantly higher proliferating effect at 9 days after operation （P〈0.05）. TGF-β1 and IGF-Ⅰ can promote the proliferation of the HSF, respectively, and the combinations of TGF-β1 and IGF-Ⅰat their optimal concentrations have better proliferating effects than the single growth factor.