表皮生长因子对人眼Müller细胞增生迁移的影响及其作用机制  

Epidermal growth factor-evoked proliferation and migration of human Müller cell

作　　者：陈春丽[1] 申焕君[1] 张宗端[1] 周仲楼[1] 陈晓燕[1] 闫东升[1] 宋宗明[1] 

机构地区：[1]温州医学院附属眼视光医院,325027

出　　处：《中华实验眼科杂志》2011年第5期444-449,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：浙江省自然科学基金项目（Y206706）

摘　　要：背景研究证实表皮生长因子（EGF）可促进鼠视网膜Müller细胞的增生和迁移，但EGF能否促进人眼Müller细胞的增生迁移尚未见报道。目的探讨人眼Müller细胞能否自身表达和分泌EGF及EGF对Müller细胞增生迁移的影响及其作用机制。方法同一代人永生株Müller细胞系MIO—M1细胞用高糖DMEM进行培养，利用MTT比色法观察不同质量浓度EGF作用24、48、72h，不同浓度CoCl2作用12h和24h，加入导致Müller细胞半数致死量的CoCl，浓度前2h加入不同质量浓度EGF的Müller细胞增生情况；Transwell小室观察正常及CoCl2缺氧条件下用不同质量浓度EGF作用24、48、72h后Müller细胞的迁移情况；采用ELISA法观察Müller细胞表达和分泌EGF的情况；通过Westernblot法检测体外不同条件培养下EGF对ERK1/2及Akt信号转导通路的作用。结果正常培养条件下，不同质量浓度的EGF作用后Mfiller细胞的吸光度（A570）值的总体比较差异有统计学意义（F=123．765，P=0．000），100mg／LEGF促Müller细胞增生作用最强，为对照组的1．6倍，10mg／LEGF促Müller细胞迁移的作用最强，为对照组的4．5倍。各EGF组Müller细胞的A570，值均明显高于对照组，差异有统计学意义（P〈O．01）。缺氧条件下培养，Müller细胞半数致死量的CoCl2浓度为600nmol／L，提前2h加入10～100阻g／LEGF后，EGF对抗缺氧所致Muller细胞的损伤作用最明显。700nmol／LCoCl2致Müller细胞损伤80％时其自身低分泌7．8ng／LEGF。10～100mg／LEGF作用Müller细胞促增生迁移作用信号最明显，600nmol／ LCoCl2作用Müller细胞24h后ERKl／2及Akt信号强度减弱，提前2h加入外源性EGF后，ERK1／2及Akt两条信号通路最明显。结论EGF能以剂量依赖的方式促进体外培养的Müller细胞增生及迁移。正常培养条件下的Müller细胞自身不表达，缺氧条件下Müller细胞自身可少量分泌EGF。EGF可能通过ERK1／2及Akt信号转Background Researches demonstrated that epidermal growth factor （EGF） can promote the proliferation and migration of Müller ceils of rat. However, whether EGF plays the role on human Müller cells is unknown. Objective Present study was to address if Müller ceils express and secret EGF and explore the effects of EGF on the proliferation and migration of Mtiller ceils under the normal and low oxygen conditions. Methods Human Müller cell line MIO-M1 ceils were cultured and incubated in DMEM with high glucose and different concentrations of EGF in the presence or absence of varied amounts of CoC12 for indicated time points. The proliferation and migration of Mailer cells were analyzed with MTT and Transwell assay respectively. The expression and secretion of EGF in Mailer cells were measured by using ELISA. Western blot was performed to detect and assess the protective role of ERK1/2 and Akt signal pathway under the presence of EGF. Results EGF stimulated the proliferation and migration of Mtiller cells in a concentration-dependent manner with a significantly different A570 values in the Muller cells among the various groups （ F = 123. 765, P = 0. 000）. The maximum proliferation rate of Muller cells was found in 100 mg/L EGF group with the 1.6 times of elevation more than the free-EGF cells. Also,the 4.5 times of increase was reached in migration rate in 10 mg/L EGF group more than free-EGF cells. In hypoxia condition, the median lethal concentration of CoC12 was 600 nmol/L. After pretreated with 10-100 mg/L EGF, the A570 values of Müller cells were obviously elevated with the statistical difference among various concentrations of EGF groups （F = 22. 900,P = 0. 000）. The ELISA results showed that MOiler cells secreted 7.8 μg/L EGF in the presence condition of 700 nmol/L CoC12. Western blot revealed that the presence of CoC12 reduced the expression of ERK1/2 and Akt in Müller eells,however,pretreatment with EGF antagonized the harmful effect of CoC12 on Müller cells. Conclusion EGF can induce the p

关 键 词：表皮生长因子 MÜLLER细胞 增生 迁移 缺氧 信号传导通路 增生性玻璃体视网膜病变 

分 类 号：R77[医药卫生—眼科]