生长因子对培养的胚胎视网膜神经细胞增殖与凋亡的影响  

Effect of growth factors on proliferation and apoptosis of cultured fetal human retinal cells

作　　者：刘文文[1] 徐萍[1] 陈霞芳[1] 黄倩[1] 

机构地区：[1]上海市第一人民医院中心实验室,200080

出　　处：《中华眼底病杂志》2003年第2期113-116,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金资助项目 (39770 2 4 2 )

摘　　要：目的　探讨表皮生长因子 ( epidermal growth factor,EGF)、成纤维细胞生长因子 ( fibroblastgrowth factor,FGF)及胎牛血清对体外培养的人胚胎视网膜神经细胞生长、增殖、分化和凋亡等的影响及可能的机制。　方法　采用胎牛血清培养人胚胎视网膜神经细胞 ,加入或不加入 EGF、FGF。通过神经元特异稀醇化酶 ( neuron specific enolase,NSE)、视网膜神经节细胞特异的 Thy1.1抗体、胶质纤维酸性蛋白( glial fibrillary acidic protein,GFAP)免疫组织化学染色、倒置显微镜及扫描电镜观察鉴定细胞成分 ;细胞生长曲线及溴脱氧尿苷 ( bromodeoxyuridine,Brd U)掺入测定细胞增殖率 ;原位末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸切口末端标记 ( Tdt- mediated d UTP nick end labelling,TUNEL)检测细胞凋亡 ;c- fos、c- jun及 bcl- 2、bax免疫组织化学染色判断转录调控因子及细胞凋亡调控因子表达水平 ,计数阳性细胞并进一步作统计分析。　结果　经过 EGF、FGF培养的人胚胎视网膜细胞总数增加 ,细胞倍增时间缩短 ,Brd U掺入率增加 ,凋亡细胞数减少 ,其中 NSE、Thy1.1阳性细胞数明显增加。同时 ,c- fos、c- jun及 bcl- 2阳性细胞数也增加。　结论　生长因子 EGF、FGF可通过上调转录因子 c- fos、c- jun及细胞凋?ObjectiveTo investigate the effects of epidermal growth factor (EGF),fibroblast growth factor(FGF), and bovine serum on proliferation and apoptosis of the cultured fetal human retinal cells.MethodsEGF and FGF were added or not to the medium of fetal human retinal cells cultured by bovine serum in vitro. The number of cells, bromodeoxyuridine(BrdU) incorporation and Tdt-mediated dUTP nick end labelling(TUNEL) were detected to determine the proliferation and apoptosis. Immunohistochemical staining of neuron specific enolase(NSE), Thy1.1, glial fibrillary acidic protein(GFAP) and scan electromicroscopy were performed to identify cell components. The expression of transcription factor c-fos, c-jun and apoptosis regulation factor bcl-2 and Bax were examined by immunohistochemical staining to explore the underlying mechanism.ResultsThe increased number of NSE and Thy1.1 positive cells and BrdU incorporation, and decreased apoptotic cells were found in the groups treated with EGF and FGF. Meanwhile, the up-regulation of c-fos, c-jun and bcl-2 were also found.ConclusionEGF and FGF can promote the survival and proliferation of cultured retinal cells by up-regulating the expression of c-fos, c-jun and bcl-2.

关 键 词：生长因子 细胞培养 胚胎 视网膜 神经细胞 细胞增殖 细胞凋亡 免疫组织化学 

分 类 号：R774.1[医药卫生—眼科]