人表皮生长因子对牛角膜内皮细胞增殖和细胞周期的影响

目的观察表皮生长因子(EGF)对牛角膜内皮细胞增殖和细胞周期的影响。方法采用体外培养的牛角膜内皮细胞,在培养液中分别添加不同浓度的人表皮生长因子(hEGF),加药后第3、7天采用MTT法,在酶标仪492nm波长处测定吸光度值观测细胞增殖,并记录细胞形态变化。将体外培养的牛角膜内皮细胞分为二组,常规DMEM培养液组和含50ng/mLEGF的DMEM培养液组,培养3、7天后,应用流式细胞仪检测细胞周期各阶段G1、S、G2期分布。结果与对照组相比,不同浓度EGF组对牛角膜内皮细胞增殖有促进作用且与浓度相关,其中10ng/mL和100ng/mL组作用强于1ng/mL组。EGF加入后3d,对照组S期细胞24.5%,G2-M期细胞0.08%,加药组S期细胞24.6%,G2-M期细胞0.06%。与对照组相比较,各期细胞分布大致相等,无显著差异。加入7d后对照组S期细胞20.8%,G2-M期细胞0.41%,加药组S期细胞18.2%,G2-M期细胞1.55%,细胞分布发生明显变化。结论EGF促进体外培养的牛角膜内皮细胞增殖,并使体外培养的牛角膜内皮细胞的细胞周期发生变化,S期细胞比例下降。

Effects of Epidermal Growth Factor on Cell Proliferation and Cell Cycle of Cultured Bovine Corneal Endothelial Cells in vitro

Objective To observe the effects of epidermal growth factor (EGF) on cell proliferation and cell cycle of cultured bovine corneal endothelial cells. Methods Bovine corneal endothelial cells were cultured with different concentrations of hEGF (1, 10 and 100 ng/mL). MTT test was performed to evaluate the cell proliferation. The bovine corneal endothelial cells were divided into two groups:control group (cultured in DMEM) and EGF-stimulated group (cultured in DMEM with EGF). Flow cytometry was performed to determine the cell cycle phases on the third and seventh day. Results Compared with the control, EGF enhanced the cell proliferation in a dose-related response. 10 ng/mL and 100 ng/mL EGF were much more effective than 1 ng/mL.On the third day, S phase cells accounted for 24.5% and G_2-M phase cells 0.08% in the control group,while 24.6% and 0.06%, respectively in the EGF-stimulated group. However, on the seventh day, those came to 20.8% and 0.41% in the control group, and 18.2% and 1.55% in the EGF-stimulated group,indicating a significant change in the cell cycle (P< 0.01). Conclusion EGF stimulates the proliferation of bovine corneal endothelial cells in vitro and influence the cell cycle as well.