摘要
目的 观察可见光诱导的培养人视网膜色素上皮 (RPE)细胞凋亡中 ,RPE细胞内碱性成纤维细胞生长因子 (bFGF)、成纤维细胞生长因子受体 1(FGFR1)、B细胞淋巴瘤 /白血病 2基因(bcl 2 )及caspase 3的表达变化。方法 建立可见光诱导培养的人RPE细胞凋亡的模型 ,通过细胞免疫组织化学、逆转录聚合酶链反应 (RT PCR)、酶联免疫测定等方法 ,对培养的人RPE细胞内bFGF和其受体FGFR1的mRNA表达变化、Bcl 2表达改变以及caspase 3的活性变化进行了检测。 结果(1)光照后 6及 12h ,培养的人RPE细胞胞质内Bcl 2蛋白的表达有一定的下降趋势 ,但与无光照组之间差异无显著性意义 (P >0 0 5 )。光照后 2 4h ,Bcl 2蛋白表达下降明显 (P <0 0 1)。bcl 2mRNA表达与上述蛋白表达变化相似 ,但早在光照 12h后已能检测到其表达下降。 (2 )光照后 6hbFGF及FGFR1的表达尚无明显改变 (P >0 0 5 ) ,随着时间延长 ,其表达增加的差异具有显著意义(P <0 0 5 )。(3)光照后caspase 3活性随时间延长而明显增加 ,与未光照时比较 ,光照结束后 6、12及2 4h活性差异有显著意义 (P <0 0 5 )。 12和 2 4h间差异无显著意义 (P >0 0 5 )。结论 可见光照诱导培养的人RPE细胞凋亡时 ,RPE细胞内bcl 2表达下降 ,内源性bFGF及FGFR1上调 ,caspase 3?
Objective To assess the expression of bFGF,FGFR1,Bcl-2 and caspase-3 in human RPE cells exposed to visible light in vitro. Methods Cultured human RPE cells were exposed to visible light [(2 000±500) Lux] for 12 hours . Immunocytochemical staining , enzyme-linked assays and RT-PCR were used to determine the expression of bFGF and its receptor FGFR1, Bcl-2 as well as the activity of caspase-3. Results (1) Bcl-2 was detected in normal cultured human RPE cells. No significant change was found at 6 or 12 hours after light exposure ( P >0.05).But 24 hours after exposure,a significantly decreasing of Bcl-2 expression was observed ( P <0.01).The changes in expression of Bcl-2 mRNA paralleled to the changes in its protein, except that differences could be detected earlier viz. 12 hours after exposure. (2) The upregulation of bFGF and FGFR1 mRNA expression occurred with longer period of light exposure ( P <0.05). (3) The upregulation of caspase-3 activity was detected 6 hours after light exposure, and was more significant over 6 hours and 12 hours ( P <0.05). No significant difference was found between 12 hours and 24 hours exposure ( P >0.05). Conclusions Visible light exposure over threshold causes upregulation of endogenous bFGF and FGFR1, downregulation of Bcl-2 and activation of caspase-3 in human RPE cells.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2003年第11期655-659,共5页
Chinese Journal of Ophthalmology
基金
国家自然科学基金资助项目 (3 9770 788)
