转化生长因子β_2对牛角膜内皮细胞增生的影响  

Inhibitory effect of transforming growth factor-β_2 on the proliferation of bovine corneal endothelial cells in vitro

作　　者：刘洋[1] 张明昌[1] 黄渝侃[1] 姜冬玲[1] 张红旭[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,武汉430022

出　　处：《眼科研究》2009年第6期502-505,共4页Chinese Ophthalmic Research

摘　　要：目的体外培养牛角膜内皮细胞(CECs),观察不同质量浓度的外源性转化生长因子β2(TGF-β2)对牛CECs的增生抑制作用。方法0.1、1、10、100μg/L的TGF-β2作用于体外培养的牛CECs,于24、48、72h观察不同质量浓度的TGF-β2对牛CECs的影响。采用CCK-8检测法测定各孔吸光度(A值),采用流式细胞检测仪对各组细胞周期进行检测。结果0.1、1、10μg/L的TGF-β2作用后各时间点,均能使牛CECs的吸光度发生改变。在同一时间点,0.1μg/L、1μg/L的TGF-β2抑制效果最为显著。各质量浓度的TGF-β2对牛CECs作用48h,其吸光度改变最明显。48h时,0.1μg/L、1μg/L的TGF-β2对牛CECs的增生抑制作用明显。各质量浓度组作用48h后,0.1μg/L、1μg/L组牛CECs的G0周期细胞所占比例与阴性对照组比较,差异均有统计学意义,而10μg/L、100μg/L组的牛CECs的G0期细胞百分比与阴性对照组比较,差异均无统计学意义。结论0.1μg/L、1μg/L的TGF-β2作用24、48、72h对牛CECs均具有明显的增生抑制作用。Objective Human corneal endothelial cell has little capacity of cell division. Endothelial repair is limited to enlargement and sliding of existing cells, and failure of endothelial function leads to cornea/ edema. Present study was to observe the inhibitory effect of transforming growth factor-β2 （TGF-β2 ） on the bovine corneal endothelial cells. Methods The corneal endothelial cells were isolated from novel bovine and digested and cultured in DMEM containing 10% fetal bovine serum. The second or the third passages of the bovine corneal endothelial cells were treated with different concentrations of TGF-β2 （0. 1,1, 10,100 μg/L）. The biologic activities of TGF-β2 in different time period and different doses of TGF-β2 group were detected with CCK-8. The flow cytometer was used to evaluate the cell cycle of negative control group and TGF-β2-treated groups. Results The cultured cells presented the confluence in the 72nd hour. The A value of the cultured cells by CCK-8 assay was significantly decreased in 0. 1,1,10 μg/L groups. The A value was most significantly decreased in all groups in the 48th hour. The inhibition effects were most significant in 0. 1 μg/L and 1 μg/L groups in the same time group（ Fgroup = 17. 916 ,P 〈0.05 ;Ftime = 22. 551, P〈0.05）.The cell cycle of the negative control group and TGF-β2 groups were examined by the Flow cytometer （FCM） 48 hours after TGF-β2 treatment. GO cells in 0. 1 μg/L and 1 μg/L TGF-I32 groups were significantly increased in comparison with negative control group（ P 〈 0. 05 ）. Conclusion TGF-β can inbihit the proliferation of bovine corneal endothelial cells from 24 through 72 hours at the concentration of 0. 1 μg/L and 1 μg/L.

关 键 词：TGF-Β2 角膜内皮细胞 细胞培养 流式细胞技术 

分 类 号：R772[医药卫生—眼科]