碱性成纤维细胞生长因子对可见光诱导的人视网膜色素上皮细胞凋亡的影响  

The effect of bFGF on apoptosis of cultured human retinal pigment epithelial cells exposed to visible light

作　　者：周咏东[1] 严密[1] 张军军[1] 

机构地区：[1]四川大学华西医院眼科

出　　处：《中华眼底病杂志》2003年第1期24-28,共5页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金资助项目 (39770 788)

摘　　要：目的　观察外源性碱性成纤维细胞生长因子 (basic fibroblast growth factor,b FGF)对可见光诱导培养的人视网膜色素上皮 (retinal pigment epithelium,RPE)细胞凋亡的影响。　方法　以 (2 0 0 0±5 0 0 ) lx白色荧光灯照射培养的人 RPE细胞 ,利用荧光素标记的连接素 V/碘化丙锭 (annexin V- fluores-cein isothiocyanate/ propidium iodium,Annexin V- FITC/ PI)双染色流式细胞测定、细胞免疫组织化学、逆转录聚合酶链反应 (reverse transcriptional polymerase chain reaction,RT- PCR)、酶联免疫测定等方法 ,检测加入不同浓度的 b FGF后 ,培养的 RPE细胞的凋亡及细胞内 b FGF、成纤维细胞生长因子受体 1(fibrob-last growth factor receptor1,FGFR1)、B细胞淋巴瘤 /白血病 - 2 (B- cell lymphom a/ leukemia- 2 ,bcl- 2 )基因及半胱天冬酶 - 3(caspase- 3)的变化。　结果　 (1)分别加入外源性 b FGF10、2 0 ng/ ml时 ,RPE细胞凋亡比单纯光照组明显减少 (P<0 .0 5 ) ;(2 )加入 b FGF 10 ng/ ml组的 bcl- 2蛋白表达比单纯光照组明显增加(P<0 .0 1) ,而与无光照组之间差异无显著性意义 (P>0 .0 5 ) ;(3)光照后 bcl- 2 m RNA表达比无光照组下降 ,但随着加入 b FGF浓度的增高 ,bcl- 2 mObjective To observe the effect of exogenous basic fibroblast growth factor (bFGF) on apoptosis of cultured human retinal pigment epithelial (RPE) cells exposed to visible light,and determine the role of bFGF, fibroblast growth factor receptor 1 (FGFR1),bcl 2 and caspase 3. Methods (2000± 500) lx cold white light was used. Exogenous bFGF was utilized during culture. Annexin annexin V fluorescein isothiocyanate/propidium iodium (V FITC/PI) labeling,flow cytometry, Immunocytochemical staining, enzyme associated absorb examing and reverse transcriptional polymerase chain reaction (RT PCR) were used to determine the apoptosis, the expression levels of bFGF, FGFR1, bcl 2, as well as the activity of caspase 3. Results No protective effect of bFGF was observed under the concentration 5 ng/ml. A significant inhibition of apoptosis was found in 10 ng/ml and 20 ng/ml groups ( P< 0 05). The upregulation of bcl 2 was observed in bFGF (10 ng/ml, 20 ng/ml) pro treated groups ( P< 0 01).Compared to no light exposure group,all light exposure groups (including bFGF pro treated) had higher endogenous bFGF and FGFR1 levels ( P< 0 05), and the increase was concentration dependent.The bFGF and FGFR1 levels were higher in exogenous bFGF applied (>5 ng/ml) groups than light exposure groups ( P< 0 05). The caspase 3 activity was significantly inhibited in bFGF (10 ng/ml) pro treated groups. Conclusions Human RPE cells exposed to visible light were rescued by application of exogenous bFGF in vitro.The probable protective mechanism of bFGF partly is directly binding to FGFR1 or potentiating endogenous bFGF autocrine loop,to upregulate bcl 2 and to inhibit caspase 3 activation.

关 键 词：视网膜 色素上皮 细胞凋亡 外源性碱性成纤维细胞生长因子 CASPASE类 

分 类 号：R774.1[医药卫生—眼科]