bFGF、EGF和NGF对人角膜内皮细胞生长调控的实验研究

目的：探讨碱性成纤维细胞生长因子（Basicfibroblastgrowthfacfor，bFGF）、表皮细胞生长因子（Epidermalgrowthfactor，EGF）和神经细胞生长因子（Nervegrowthfactor，NGF）对体外培养的人角膜内皮细胞的生长调控作用。方法：将相同数量的人角膜内皮细胞接种于96孔板。加入浓度分别为0ng／ml、1ng／ml、3ng／ml、10ng／ml、30ng／ml、100ng／ml的EGF、bFGF和NGF进行培养。5天后MTT法用检测增殖情况。结果：在0ng／ml、1ng／ml、3ng／ml、10ng／ml、30ng／ml、100ng／ml浓度下bFGF组的平均OD值分别为：0．224±0．045、0．239±0．040、0．262±0．0342、0．278±0．0319、0．281±0．0324、0．260±0．0310。EGF组的平均OD值分别为：0．228±0．0304、0．245±0．0418、0．267±0．0454、0．275±0．0347、0．271±0．0449、0．250±0．0253。NGF组的平均OD值分别为：0．216±0．0187、0．228±0．0226、0．231±O．0225、0．242±0．0279、0．245±0．0294、0．247±0．0349。结论：bFGF在30ng／ml范围内对内皮细胞生长有促进作用，并具有剂量依赖性。高于100ng／ml时促生长作用降低。EGF在10ng／ml范围内对内皮细胞生长有促进作用，并具有剂量依赖性。高于30ng／ml时促生长作用降低。NGF本次实验剂量范用内对角膜内皮细胞生长无明显作用。

A Study on Effect of bFGF、EGF and NGF on Growth of Cultured Human Corneal Endothelial Cells

PURPOSE: To investigate effect of bFGF, EGF and NGF on growth of cultured human corneal endothelial cells. METHODS: Cultured human corneal endothelial cells were seeded into individual wells of 96-well tissue culture plate with the same culture media containing separately bFGF, EGF or NGF with a serial of concentrations of 0 ng/ml, 1 ng/ml, 3 ng/ml, 10 ng/ml, 30 ng/ml and 100 ng/ml and then cultured for 5 days. Then MTT method was used to detect the growth of the cells. RESULTS: The averaged OD values of the cell wells containing bFGF with a serial of concentrations of 0 ng/ml, 1 ng/ml, 3 ng/ml, 10 ng/ml, 30 ng/ml and 100 ng/ml were 0.224 +/- 0.045, 0.239 +/- 0.040, 0.262 +/- 0.0342, 0.278 +/- 0.0319, 0.281 +/- 0.0324, 0.260 +/- 0.0310. The averaged OD values of EGF group and NGF group were separately 0.228 +/- 0.0304, 0.245 +/- 0.0418, 0.267 +/- 0.0454, 0.275 +/- 0.0347, 0.271 +/- 0.0449, 0.250 +/- 0.0253 and 0.216 +/- 0.0187, 0.228 +/- 0.0226, 0.231 +/- 0.0225, 0.242 +/- 0.0279, 0.245 +/- 0.0294, 0.247 +/- 0.0349. CONCLUSION: bGFG can promote the growth of human corneal endothelial cells in a dose dependent manner while with concentration lower than 30 ng/ml. bFGF with concentration that is higher than 100 ng/ml will weaken this effect. EGF can also promote proliferation of human corneal endothelial cells demonstrating a linear dose dependent effect when its concentration is lower than 10 ng/ml and this effect decreased when its concentration was higher than 30 ng/ml. NGF showed no effect on the growth of human cornea endothelial cells in this study.