Wang Zhi-Xin, Sun Xu-Guang
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Capital University of Medical Sciences, Beijing, China.
Zhonghua Yan Ke Za Zhi. 2007 Oct;43(10):928-31.
To study the effect of nerve growth factor (NGF) on the proliferation and the expression of MUC5AC gene in cultured human conjunctival goblet cells.
The presence of high affinity receptors of nerve growth factor (TrkA) in cultured human conjunctival goblet cells was identified with immunocytochemical analysis. Proliferation of goblet cells was determined by MTT proliferation assay after exposure to different concentrations of NGF, (20, 40, 60 and 80 ng/ml) in serum-free RPMI-1640 culture medium (RPMI) for 1, 3 and 5 days. The effect of NGF on gene expression of MUC5AC of goblet cells was determined by RT-PCR after exposing cells to NGF (80 ng/ml in serum-free RPMI) for 5 days.
Human conjunctival goblet cells in subculture exhibited positive staining for TrkA. After exposure to NGF, the differences in absorption value of MTT test of goblet cells cultured with different levels of NGF (F = 12.766, P = 0.000) and at different time points (F = 892.406, P = 0.000) were statistically significant, but the difference in interaction effects was not significant. MUC5AC gene expression of NGF group was higher than that of the control group (t = 4.963, P = 0.000).
NGF receptors (TrkA) are present in human conjunctival goblet cells. NGF causes a dose-dependent increase in the proliferation of goblet cells. The expression of MUC5AC gene in goblet cells is upgraded by NGF.
研究神经生长因子（NGF）对培养的人结膜杯状细胞增殖及MUC5AC基因表达的影响。
采用免疫细胞化学分析法鉴定培养的人结膜杯状细胞中神经生长因子高亲和力受体（TrkA）的存在情况。在无血清的RPMI-1640培养基（RPMI）中，将杯状细胞暴露于不同浓度（20、40、60和80 ng/ml）的NGF中1、3和5天，通过MTT增殖试验测定杯状细胞的增殖情况。将细胞暴露于无血清RPMI中80 ng/ml的NGF 5天后，采用RT-PCR法测定NGF对杯状细胞MUC5AC基因表达的影响。
传代培养的人结膜杯状细胞TrkA染色呈阳性。暴露于NGF后，不同水平NGF培养的杯状细胞MTT试验吸光度值在不同时间点的差异有统计学意义（F = 12.766，P = 0.000），但交互作用差异无统计学意义。NGF组MUC5AC基因表达高于对照组（t = 4.963，P = 0.000）。
人结膜杯状细胞中存在NGF受体（TrkA）。NGF可使杯状细胞增殖呈剂量依赖性增加。NGF可使杯状细胞中MUC5AC基因表达上调。
