Pan Jin-Shun, Zhu Huai-Jun, Zhang Bei, Li Hua, Yan Hong, Wang Bin
Department of Pharmacology, Nanjing Medical University, Nanjing, People's Republic of China.
J Ocul Pharmacol Ther. 2006 Apr;22(2):103-9. doi: 10.1089/jop.2006.22.103.
The time course changes of basic fibroblast growth factor (bFGF) expression induced by hypoxia and the effects of genistein on hypoxia-induced bFGF expression in the human retinal pigment epithelium (RPE) cells were studied.
The bFGF mRNA expression was examined by reverse transcription polymerase chain reaction. The bFGF protein expression was detected by Western blot.
Hypoxia significantly increased bFGF mRNA expression. The maximal level detected at 24 h was approximately two times that at the start of treatment. With pretreatment of genistein (10, 20, 50, 100, and 200 microM) for 30 min, the elevated expression of bFGF mRNA was suppressed in a concentration-dependent manner. bFGF mRNA expression was reduced to 30.4% by 200 microM of genistein when compared with that untreated with genistein. Hypoxia treatment also remarkably increased the expression of bFGF protein. At 24 h after hypoxia, when the highest expression of bFGF protein was observed, it was approximately two times as much as that at the start of treatment. Genistein (10, 20, 50, 100, and 200 microM) could also suppress bFGF protein expression in a concentration-dependent manner. The highest suppression was observed when exposed to 200 microM of genistein, which was 43% of control.
These results suggested that suppression of bFGF expression in RPE cells might partly account for the inhibitive effect of genistein on retinal neovascularization in vivo.
研究缺氧诱导人视网膜色素上皮(RPE)细胞碱性成纤维细胞生长因子(bFGF)表达的时间进程变化以及染料木黄酮对缺氧诱导的bFGF表达的影响。
采用逆转录聚合酶链反应检测bFGF mRNA表达。通过蛋白质印迹法检测bFGF蛋白表达。
缺氧显著增加bFGF mRNA表达。在24小时检测到的最高水平约为治疗开始时的两倍。用染料木黄酮(10、20、50、100和200微摩尔)预处理30分钟后,bFGF mRNA的升高表达以浓度依赖的方式受到抑制。与未用染料木黄酮处理相比,200微摩尔染料木黄酮使bFGF mRNA表达降低至30.4%。缺氧处理也显著增加bFGF蛋白的表达。在缺氧后24小时,观察到bFGF蛋白表达最高时,约为治疗开始时的两倍。染料木黄酮(10、20、50、100和200微摩尔)也能以浓度依赖的方式抑制bFGF蛋白表达。当暴露于200微摩尔染料木黄酮时观察到最高抑制率,为对照的43%。
这些结果表明,抑制RPE细胞中bFGF的表达可能部分解释了染料木黄酮对体内视网膜新生血管形成的抑制作用。