姜黄素在抑制紫外线B诱导的人视网膜色素上皮细胞DNA氧化损伤和凋亡中的作用  

Inhibitory effect of curcumin on ultraviolet-B-induced DNA oxidative damage and apoptosis in human retinal pigment epithelial cells

作　　者：汪涛 何琎 刘亚 卢萍 张梦瑜 刘建军 WANG Tao;HE Jin;LIU Ya;LU Ping;ZHANG Mengyu;LIU Jianjun(Department of Ophthalmology,Suzhou Science and Technology Town Hospital,Suzhou 215000,Jiangsu Provience,China)

机构地区：[1]苏州科技城医院眼科,江苏省苏州市215000

出　　处：《眼科新进展》2023年第7期530-535,共6页Recent Advances in Ophthalmology

摘　　要：目的 探讨姜黄素在紫外线B(UVB)诱导的人视网膜色素上皮(RPE)细胞DNA氧化损伤和凋亡中的作用机制。方法 将人视网膜色素上皮细胞株(ARPE-19)随机分为空白对照组、二甲基亚砜(DMSO)组、姜黄素组、UVB照射组、UVB+DMSO组和UVB+姜黄素组。CCK-8实验检测姜黄素对UVB照射前后ARPE-19细胞活力的影响;免疫荧光检测DNA氧化损伤标志蛋白磷酸化组蛋白H2AX(γH2AX)的定位;Western blot检测γH2AX、凋亡相关蛋白[B淋巴细胞瘤-2基因(BCL-2)和BCL-2相关X蛋白(BAX)]的表达变化;使用X-rosamine衍生物(Mito-Tracker Red CMXRos)和异硫氰酸荧光素(FITC)标记的钙离子依赖的磷脂结核蛋白Annexin V(Annexin V-FITC)来检测UVB诱导的ARPE-19细胞线粒体膜电位变化和细胞凋亡情况。结果 与空白对照组相比,DMSO组DMSO处理24 h后的ARPE-19细胞活力无明显改变(P>0.05)。与DMSO组相比,姜黄素组采用15μmol·L^(-1)姜黄素处理24 h后ARPE-19细胞活力轻度升高(P<0.05),表明15μmol·L^(-1)姜黄素对ARPE-19细胞无毒性作用,后续实验采用该浓度处理细胞。与空白对照组相比,UVB照射组ARPE-19细胞活力降低,γH2AX和BAX蛋白表达水平升高,BCL-2蛋白表达水平降低,线粒体膜电位降低,细胞凋亡增加(均为P<0.05)。与UVB照射组相比,UVB+DMSO组ARPE-19细胞活力,γH2AX、BAX以及BCL-2蛋白表达水平均无明显变化(均为P>0.05),线粒体膜电位和细胞凋亡均无明显改变(均为P>0.05)。与UVB+DMSO组相比,UVB+姜黄素组ARPE-19细胞活力升高,γH2AX和BAX蛋白表达水平降低,BCL-2蛋白表达水平升高,线粒体膜电位升高,细胞凋亡减少(均为P<0.05)。结论 姜黄素能减轻UVB诱导的RPE细胞DNA氧化损伤和凋亡,有望为姜黄素在年龄相关性黄斑变性中的作用机制研究提供新的思路。Objective To investigate the effects of curcumin on DNA oxidative damage and apoptosis in human retinal pigment epithelial(RPE)cells induced by ultraviolet-B(UVB).Methods Human RPE cell lines(ARPE-19)were divided into the blank control(BC)group,dimethyl sulfoxide(DMSO)group,curcumin group,UVB irradiation group,UVB+DMSO group,and UVB+curcumin group.The effect of curcumin on the activity of ARPE-19 cells before and after UVB irradiation was assessed by CCK-8 assay.The H2AX phosphorylation(γH2AX),a marker of DNA oxidative damage,was localized by immunofluorescence.The expression levels ofγH2AX and apoptosis-related proteins,including B-cell lymphoma 2(BCL-2)and Bcl-2-associated X protein(BAX),were measured by Western blot.X-rosamine derivative Mito-Tracker Red CMXRos and Annexin V labeled by fluorescein isothiocyanate(Annexin V-FITC)were used to detect UVB-induced mitochondrial membrane potential changes and apoptosis in ARPE-19 cells.Results Compared with the BC group,ARPE-19 cells in the DMSO group showed no significant changes in viability after being treated with DMSO for 24 h(P>0.05).Compared with the DMSO group,the viability of ARPE-19 cells after being treated with 15μmol·L^(-1) curcumin for 24 h in the curcumin group increased slightly(P<0.05),indicating that curcumin at this concentration had no cytotoxicity,and it was used in the subsequent experiments.Compared with the BC group,ARPE-19 cell viability,BCL-2 level,and mitochondrial membrane potential in the UVB irradiation group decreased,whileγH2AX and BAX levels and cell apoptosis increased(all P<0.05).Compared with the UVB irradiation group,ARPE-19 cell viability,γH2AX,BAX and BCL-2 levels,mitochondrial membrane potential,and cell apoptosis in the UVB+DMSO group showed no significant changes(all P>0.05).Compared with the UVB+DMSO group,ARPE-19 cell viability,BCL-2 level,and mitochondrial membrane potential increased,whileγH2AX and BAX levels and cell apoptosis decreased in the UVB+curcumin group(all P<0.05).Conclusion Curcumin can reduce DNA oxida

关 键 词：姜黄素 紫外线B 人视网膜色素上皮细胞 DNA氧化损伤 细胞凋亡 

分 类 号：R776[医药卫生—眼科]