原代培养兔晶状体上皮细胞的环境和细胞状态的变化

Change of status of primary culture rabbits' lens epithelial cells

目的:探讨体外原代培养的兔晶状体上皮细胞(lens epithelial cells,LEC)分泌活性转化生长因子β1(transforming growth factorβ1,TGF-β1)和异常表达α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的状况变化方法:体外原代培养兔眼LEC,当细胞接近融合时转板培养,观察细胞转板培养后1,6,12d的状态变化,并采用ELISA法检测各时项培养液上清中TGF-β1的水平,免疫细胞化学和计算机图像分析统计各时项细胞中α-SMA的表达水平。结果:转板培养12d的细胞活性TGF-β1的水平明显高于转板培养1和6d的水平(均P<0.05),同时转板培养12d的细胞表达α-SMA的水平分别明显高于转板培养1和6d的水平(P<0.05);而且α-SMA的表达量随着TGF-β1水平的升高而升高,两者高度正相关(r=0.995,P<0.01)。结论:原代培养的LEC活性TGF-β1逐渐增多,向表达有α-SMA的肌成纤维细胞转分化。

AIM： To observe the change of secretion of active transforming growth factor β1 （TGF-β1） and abnormal expression of α-smooth muscle actin （α-SMA） of primarily cultured rabbits＇ lens epithelial cells in vitro. METHODS： Rabbits＇ lens epithelial cells were primarily cultured in vitro. When cell fusion, it was turned to boards, After 1,6and 12 days, we observed the change of cells state, tested the levels of TGF-β1 in every medium by ELISA, and tested the levels of α-SMA in cells by immuocytochemistry and computer image analysis, RESULTS： The levels of active TGF-β1 and α-SMA of the cells after 12 days were significantly higher than the levels of the cells after 1 and 6 days（P〈0.05）, α-SMA increased accompanying to the increasing level of TGF-β1 and the both were highly positive correlated（ r= 0.995, P〈 0,001 ）. CONCLUSION： Primarily cultured rabbits＇ lens epithelial cells secrete more and more active TGF-β1 gradually, and with the effect of the large TGF-β1, the cells become to transdifferentiation to myofibroblasts which express α-SMA. Therefore, this study provids a good cell analogue for traumatic and after cataract.