原代培养兔晶状体上皮细胞的环境和细胞状态的变化

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。

Change of status of primary culture rabbits' lens epithelial cells

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 epi-thelial 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.