α-倒捻子素对过氧化氢诱导的人视网膜色素上皮细胞损伤的保护作用  

Protective effects of α-Mangostin on injury of human retinal pigment epithelium cells induced by hydrogen peroxide

作　　者：苏途 方圆[1] 谢平[1] 袁松涛[1] 范雯[1] 许译丹[1] 刘庆淮[1] 

机构地区：[1]南京医科大学第一附属医院眼科,中国江苏省南京市210029

出　　处：《国际眼科杂志》2015年第6期959-962,共4页International Eye Science

基　　金：国家重点基础研究发展计划(973计划No.2013CB967500;No.2011CB510200);国家自然科学基金项目面上项目(No.81170855);江苏省临床医学科技专项--重点病种的规范化诊疗研究(No.BL2014089);"六大人才高峰"D类资助项目(No.2013-WSW-015)~~

摘　　要：目的：研究α-倒捻子素对过氧化氢（H2O2）诱导的人视网膜色素上皮细胞株ARPE-19氧化损伤的保护作用。方法：分别用不同浓度的α-倒捻子素和H2O2处理ARPE-19,CCK8法检测α-倒捻子素和H2O2对ARPE-19细胞毒性作用。不同浓度的α-倒捻子素预处理ARPE-19,再用200μmol/L H2O2处理24h,观察细胞活性变化。流式细胞仪检测细胞活性氧（reactive oxygen species,ROS）水平变化,免疫印迹法（Western blot）检测NF-kB蛋白表达变化。结果：CCK8法检测结果显示：当α-倒捻子素浓度在0～12μmol/L时,ARPE-19活性无明显变化;当浓度达到16μmol/L时,细胞活性开始下降（P〈0.05）。H2O2诱导后,当α-倒捻子素浓度在0～16μmol/L之内时,α-倒捻子素预处理均可提高ARPE-19细胞活性。ROS结果表示：H2O2诱导后,ROS表达量增高（P〈0.05）;8和12μmol/Lα-倒捻子素预处理,均可有效清除H2O2诱导产生的ROS（P〈0.05）。Western blot结果显示：H2O2诱导后NF-kB蛋白表达增高（P〈0.05）,12μmol/Lα-倒捻子素预处理可以继续上调H2O2诱导后ARPE-19的NF-kB蛋白表达（P〈0.05）。结论：H2O2诱导ARPE-19细胞氧化损伤,造成细胞活性下降,ROS表达量增加,经过α-倒捻子素预处理后,可有效提高细胞活性,清除ROS,活化NF-kB。AIM： To study protective effects of α-Mangostin in human retinal pigment epithelial （RPE） cells induced by hydrogen peroxide（ H202 ）. METHODS：ARPE-19 cells were treated with different concentrations of α-Mangostin and H2O2. The effect of α- Mangostin and H2O2 respectively on cell activity was detected by CCK8. ARPE-19 cells were pretreated with different concentrations of a-Mangostin for 24h before they were administrated with 200μmol/L H2O2 for another 24h. Then the changes of cell activity were observed. The expression of reactive oxygen species （ROS） level was detected by flow cytometry（FCM） and the expression of NF-κB protein was measured by Western blot analysis. RESULTS： CCK8 examination results showed that= within 0 -12μmol/L, α-Mangostin had no damage effects on cell activity. When the concentration of 16pmol/L, cell viability began to decrease （ P〈 0.05）. And α- Mangostin pretreatment gradually increased cell viability of ARPE-19 induced by H202 when the concentrations of α-Mangostin were within 0 ~ 16μmol/L. ROS results showed, the expression of ROS level significantly increased after H202 induced （P〈 0. 05）; 8 and 12tJmol/L α- Mangostin pretreatment down-regulated ROS expression of ARPE-19 induced by H202（P〈0.05）. Western blot results showed that the expression of NF-κB protein after H2 02 induced increased （ P 〈 0. 05 ）; 12μmol/L α- Mangostin pretreatment up- regulated NF- κB of ARPE- 19 induced by H202（P〈0.05）. CONCLUSION： H202 induced oxidative damage in RPE cells by decreasing cell viability and increasing the expression of ROS level, a-Mangostin can protect RPE cells from the injury of H2 02, the mechanism may be related to the clear of ROS and the activation of NF-κB.

关 键 词：α-倒捻子素 视网膜色素上皮细胞 氧化损伤 细胞核因子-ΚB 细胞活性氧 

分 类 号：R774.5[医药卫生—眼科]