摘要
目的研究金雀异黄素(genistein,Gen)对培养的人视网膜色素上皮(retinal pig-ment epithelium,RPE)细胞的诱导凋亡作用,探讨其作用机制。方法 0、25、50、100、200μmol.L-1Gen分别作用RPE细胞24h;MTT检测细胞活性;流式细胞仪检测细胞周期,定量分析凋亡;免疫印迹技术检测细胞外调节蛋白激酶(extracellular regulated proteinkinas-es,ERK)、p38MAPK的表达及活性。结果不同浓度的Gen作用RPE细胞24h后,细胞生长明显受到抑制,抑制率分别为6.44%、14.71%、28.97%、45.75%;细胞凋亡率明显增高,分别为16.87%、19.44%、23.14%、34.81%;细胞周期表现为S期和G2/M期阻滞,并伴随G0/G1期细胞减少;p-ERK1/2表达下调,p-p38表达下调。ERK1/2的激活与Gen诱导的细胞凋亡呈负相关,p38的激活与凋亡可能也存在一定的关系。结论 Gen可以抑制人RPE细胞的增殖,并诱导其凋亡,存在量效关系,且主要是通过ERK通路发挥生物学效应。
Objective To investigate the mechanism and effects of genistein（Gen）on induced apoptosis of cultured human retinal pigment epithelium（RPE）cells.Methods The 0 μmol·L-1,25 μmol·L-1,50 μmol·L-1,100 μmol·L-1 and 200 μmol·L-1 Gen were used to treat RPE for 24 hours,respectively.MTT was used to detect cell activity,flow cytometry to detect cell cycle for quantitatively analyzing apoptosis,and immunoblotting to detect expression and activity of extracellular regulated protein kinases（ERK）and p38MARK.Results After different concentrations of Gen treating RPE for 24 hours,cell growth was obviously inhibited,inhibitive rates were 6.44%,14.71%,28.97% and 45.75%;Rates of cell apoptosis were obviously increased,and were 16.87%,19.44%,23.14% and 34.81%.Cell cycles of S stage and G2/M stage were blocked;Cells at G0/G1 stage were decreased.Expression of p-ERK1/2 and p-p38 were with down-regulation.There was negative correlation between activation of ERK1/2 and cell apoptosis induced by Gen,and activation of p38 may also have relation with apoptosis.Conclusion Gen can inhibit proliferation of RPE cells,induce cell apoptosis and there is dose-depend manner,which mainly have biological effect by ERK pathway.
出处
《眼科新进展》
CAS
北大核心
2010年第12期1123-1126,共4页
Recent Advances in Ophthalmology
基金
南京医科大学创新基金资助(编号:CX2001003)~~
