神经生长因子对人巩膜成纤维细胞生长及胶原蛋白合成的影响

目的探索神经生长因子（NGF）对培养的人巩膜成纤维细胞（HSF）生长及胶原蛋白合成的影响。方法采用MTT比色法和流式细胞仪（FCM）分析技术，观察不同质量浓度NGF（10、25、50、100、200ng／ml）对HSF增殖和细胞生长周期的影响，氯胺T法检测胶原蛋白。结果NGF能促进HSF增殖，呈剂量依赖性，质量浓度为100ng／ml时作用最明显。NGF作用后，HSF的G0～G1期百分比降低，而S期百分比显著升高，增殖指数增高。NGF对HSF的胶原合成具有明显的促进作用，并呈剂量依赖性。结论外源性NGF可以促进体外培养的HSF增殖及胶原合成。

Effects of NGF on proliferation and collagen synthesis of human scleral fibroblast cells

[Objective] To investigate the effects of nerve growth factor(NGF)on the proliferation and collagen synthesis in cultured human fetal scleral fibroblast cell(HSF).[Methods] Primary culture and subculture of HSF cells were established in vitro firstly.Cultured HSF cells were treated with NGF at various concentrations of 10,25,50,100,200 ng/ml for 48 h.Cells proliferation was measured with methylthiazolyl tetrazolium(MTT) assay and cell cycle was determined by flow cytometry(FCM).Hydroxyproline contents were measured with the chloramine T method.[Results] NGF could significantly promote the proliferation and collagen synthesis of scleral fibroblast in a dose-dependent manner with a peak action at the concentration of 100 ng/ml.The results of FCM analysis showed that percentage of scleral fibroblast in S phase was increased and that of G0~G1 was decreased when the cells were stimulated with NGF in comparison with control group.The percentage of proliferation index(PI) was increased.[Conclusion] NGF can stimulate the proliferation and collagen synthesis of scleral fibroblast in vitro in a dose-dependent manner.