ET-1、rhEGF、bFGF对培养的兔角膜内皮细胞的影响

目的 研究内皮素-1(ET-1)、重组人表皮生长因子(rhEGF)、碱性成纤维细胞生长因子(bFGF)对培养的兔角膜内皮细胞增殖能力的影响及其相互作用。方法 在体外培养的兔角膜内皮细胞中单独使用或联合应用ET-1、rhEGF、bFGF，采用MTT方法观察对细胞增殖的影响，免疫组化染色法和计算机图像分析系统检测对细胞增殖细胞核抗原(PcNA)表达的影响。结果 一定浓度ET-1促进培养的兔角膜内皮细胞的增殖，且呈剂量相关性。10pmol／L时起作用，200pmol／L时发挥最大作用。10ng／ml rhEGF、2ng／mlbFGF也促进培养的兔角膜内皮细胞的增殖。联合应用ET-1 rhEGF、ET-1 bFGF、ET-1 rhEGF bFGF，细胞吸光度(A)值明显高于单独使用相应药物时A值之和。ET-1、rhEGF、bFGF单独使用可促进培养的兔角膜内皮细胞PCNA表达，联合应用时PCNA表达平均吸光度A值明显强于单独使用相应药物时表达强度之和。结论 ET-1可作为一种生长因子，它和rhEGF、bFGF单独使用可促进培养的兔角膜内皮细胞增殖能力，联合应用时具有协同作用。

Effects of ET-1, rhEGF and bFGF on Cultured Rabbit Corneal Endothelial Cells in vitro

Objective To study the effects of endothelin-1 (ET-1), recombinant human epidermal growth factor (rhEGF) and basic fibroblast growth factor (bFGF) on proliferation of the cultured rabbit endothelial cells in vitro and their interactions. Methods Rabbit corneal endothelial cells in culture were treated with different concentrations of ET-1, and MMT assay was used to examine the effect of ET-1 on proliferation of cells after 48 h. When ET-1, rhEGF and bFGF was used alone or together, MMT assay was used to examine the effect on proliferation of cells, and immunohischemistrical staining and computer medical image analysis system were used to detect the expression of proliferating cell nuclear antigen (PCNA) of cells. Results ET-1 at the rank from 10 pmol/L to 200 pmol/L had the effect on proliferation of cultured rabbit endothelial cells in vitro in a dose-dependent manner. At the concentration of 200 pmol/L, ET-1 had the maximal effect. rhEGF at the concentration of 10 ng/ml or bFGF at the concentration of 2 ng/ml also had the effect on proliferation of the cultured rabbit endothelial cells in vitro. In ET-1 rhEGF, ET-1 bFGF and ET-1 rhEGF bFGF, the A values exceeded sum of them used alone. ET-1, rhEGF and bFGF could promote the expression of PCNA in cultured rabbit endothelial cells in vitro if used alone, and if used together, the average A value exceeded that of sum of them. Conclusion Acting as a growth factor on corneal endothelium, ET-1 has the effect on proliferation of the cultured rabbit endothelial cells in vitro like rhEGF or bFGF. And there is the synergistic action if they are used together.