缺氧时Genistein对人RPE细胞生长作用机制的初步研究

Genistein on the proliferation of RPE cells under anoxic culture

目的:研究缺氧条件下金雀异黄素(genistein,Gen)对培养人视网膜色素上皮细胞(retinalpigmentepitheliumcell,RPE)的生长影响,探讨缺氧时Gen对人RPE细胞生长的作用机制。方法:以不缺氧为对照,建立人RPE细胞的缺氧模型,分别以Gen0、50、100、200μmol/L(组)的药物浓度作用细胞;用甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)测定法测定24h细胞活力(以吸光度A值表示);用RT鄄PCR(反转录-聚合酶链反应)技术检测细胞内RasmRNA表达;用酶联免疫吸附试验技术(ELISA)检测硝基酪氨酸(3鄄Nitrotyrosine,3鄄NT)的产量;用黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(superoxidedismutase,SOD)的活力。结果:与对照组比,缺氧条件下,随着Gen浓度增高,24h3鄄NT的生成量均增加(P<0.05),SOD代偿性增高(P<0.05);与Gen0组比较,Gen50、100、200组Ras基因相应表达升高。Gen在100、200μmol浓度时,24h细胞MTT检测较Gen0A值显著升高(P<0.05),表明Gen的作用下,缺氧RPE细胞出现增殖现象。结论:Gen增强了缺氧24h的RPE细胞内的氧化应激水平,Ras基因mRNA表达增高,细胞增殖,可能同导致3鄄NT生成增多的自由基有关。

Objective: To study the effects of different concentrations of Gen o n the proliferation of RPE cells under anoxic culture conditions. Methods: Two m odels of RPE cells were set up, one under anoxic conditions and the other under normal conditions as control. The four groups of RPE cells in the first model w ere treated with Gen 0,50,100,200 μmol/L respectively. After 24 h, the A value of MTT of RPE cells was measured, and the expression of Ras mRNA was detected wi th RT-PCR. ELISA was used to detect 3-NT , and Xanthine Oxidase Enzymic method for the validity of SOD. Results: In anoxic environment, with stronger genistein , there was increased expression of Ras mRNA in the cells. Compared with the co ntrol goup there was a significantly bigger increase of 3-NT(P < 0.05),and a bet ter expression of SOD(P < 0.05). Compared with the RPE cells not treated with Ge n in the anoxic environment, the A value showed a significant increase in the RP E cells treated with Gen 100,200(P < 0.05), indicating a proliferation of RPE ce lls in 24 h. Conclusion: Gen could improve the oxidative stress of RPE cells und er anoxic conditions and lead to the increased expression of Ras mRNA and the pr oliferation of RPE cells, which might be related to the ROS that resulted 3-NT increase.