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目的 研究神经生长因子 (NGF)信号系统在雌激素调控视网膜血管内皮细胞中的作用。 方法 选用恒河猴视网膜 脉络膜血管内皮细胞系RF 6A ,RT PCR检测雌激素受体 (estrogenreceptor,ER)、NGF及其受体在视网膜血管内皮细胞中的表达。分别观察雌二醇 (estradiol,E2 )、NGF及VEGF、EGF、BDNF等对血管内皮细胞活力(MTT法 )的作用。用流式细胞术 细胞周期法检测E2 、NCF及E2 与K2 5 2a共同处理组的凋亡率。NGF抗体和选择性TrkA拮抗剂K2 5 2a研究对上述作用的阻断效应。同样的分组进行细胞划痕修复实验。 96孔板AngioMatrix胶上观察E2 及NGF对RF 6A内皮细胞管腔形成能力的影响。 结果 RF 6A在细胞外基质胶上可形成管腔样结构。RT PCR检测到ER α、NGF及NGF受体TrkA的mRNA。 1nmol L～ 1 0 0nmol LE2 可以剂量依赖性的增强细胞活力及单层细胞的划痕修复能力 ,其增强作用可部分被NGF抗体及 1 0 0nmol L的K2 5 2a选择性阻断。凋亡率检测显示各组凋亡率相近。E2 可促进RF 6A形成管腔 ,但不能被NGF抗体、TrkA阻断。 结论 除VEGF外 ,E2 还可通过NGF信号系统对视网膜血管内皮细胞活力、划痕修复起促进作用。但结果提示 ,阻断NGF信号系统不影响E2 促RF 6A形成管腔的作用。
Objective To investigate the role of nerve growth factor (NGF) signaling system in the regulation of retinal vascular endothelial cells by estrogen. Methods Rhesus monkey retinal choroidal vascular endothelial cell line RF 6A was used to detect the expression of estrogen receptor (ER), NGF and its receptor in retinal vascular endothelial cells. The effects of estradiol (E2), NGF, VEGF, EGF and BDNF on the viability of vascular endothelial cells (MTT) were observed. Flow cytometry was used to detect the apoptosis rates of E2, NCF and E2 and K2 5 2a co-treatment groups. NGF antibody and selective TrkA antagonist K2 5 2a studied the blocking effect on these effects. The same group was used for cell scratch repair experiments. The effects of E2 and NGF on the lumen formation ability of RF 6A endothelial cells were observed on 96-well AngioMatrix gel. Results RF 6A forms a luminal-like structure on the extracellular matrix gel. The mRNAs of ERα, NGF and NGF receptor TrkA were detected by RT PCR. 1nmol L ~ 100nmol LE2 can increase the cell viability and scratch repair ability of monolayer cells in a dose-dependent manner, and its enhancement can be partially blocked by NGF antibody and K2 5 2a of 100 nmol L. Apoptosis rate showed similar rates of apoptosis in each group. E2 can promote the formation of lumen of RF 6A, but can not be blocked by NGF antibody, TrkA. Conclusion In addition to VEGF, E2 can also promote retinal vascular endothelial cell viability and scratch repair through the NGF signaling system. However, the results suggest that blocking the NGF signaling system does not affect the effect of E2 on promoting RF 6A lumen formation.