摘要
目的探讨白藜芦醇(Res)对高糖所致视网膜色素上皮细胞损伤的保护作用。方法先将鉴定后的ARPE-19细胞株分为正常对照组、高糖组和高渗组培养24、48、72 h,CCK-8法检测各时间点细胞活力确定最佳作用时间;再依据上述结果将ARPE-19细胞株分为不同浓度Res治疗组及正常对照组培养48 h,CCK-8法检测各组细胞活力确定Res最佳作用浓度。随后将ARPE-19细胞株分为正常对照组、HG组(高糖)、HG+Res组(高糖+Res)培养48 h,倒置显微镜观察各组细胞生长情况和形态变化;采用化学比色法检测丙二醛(MDA)水平和超氧化物歧化酶(SOD)、谷胱甘肽(GSH-Px)活性,ELISA法检测各组血管内皮生长因子(VEGF)、色素上皮衍生因子(PEDF)水平,RT-PCR检测各组肿瘤坏死因子-α(TNF-α)mRNA、白细胞介素-1β(IL-1β)mRNA、IL-6 mRNA的水平。结果与HG组比较,HG+Res组形态学异常改变明显缓解。与HG组比较,HG+Res组SOD、GSH-Px活性降低,MDA活性增加(P<0.05);与HG组比较,HG+Res组VEGF水平降低,PEDF水平升高(P<0.05)。与正常对照组比较,HG组TNF-αmRNA、IL-1βmRNA、IL-6 mRNA水平升高(P<0.05);与HG组比较,HG+Res组TNF-αmRNA、IL-1βmRNA、IL-6 mRNA水平降低(P<0.05)。结论Res可能通过抗氧化、抑制增殖、抗炎等途径延缓高糖环境PRE细胞的损伤。
Objective To investigate the protective effect of resveratrol on the damage of retinal pigment epithelial cell caused by high glucose.Methods First,the identified ARPE-19 cell lines were divided into the control group,the high glucose group and the hyperosmotic group and cultured for 24,48,72 h,cell viability of all groups were evaluated by cell counting kit-8(CCK-8)assay at the three time points to select the most appropriate intervention time.Based on the above results,ARPE-19 cell lines were divided into different concentrations of resveratrol treatment groups and normal control group,and cultured for 48 h,the cell activity of each group was detected by CCK-8 method to determine the optimal concentration of resveratrol.Subsequently,ARPE-19 cell lines were divided into the normanl control group,the HG group(high glucose),the HG+Res group(high glucose+resveratrol)and cultured for 48 h,and cell morphology was investigated by phase contrast inverted microscope.The level of MDA and activities of SOD and GSH-Px were detected by chemical colorimetric method.The levels of VEGF and PEDF in different groups were determined by ELISA.The expression levels of TNF-αmRNA,IL-1βmRNA,IL-6 mRNA were quantified by real-time quantitative PCR(RT-PCR).Results Compared with the HG group,the abnormal morphological changes of the HG+Res group were significantly alleviated.Compared with the HG group,the SOD and GSH-Px activities in the HG+Res group decreased and the MDA level increased(P<0.05),the expression level of VEGF in the HG+Res group decreased and the expression level of PEDF increased(P<0.05).Compared with the control group,the expression levels of TNF-αmRNA,IL-1 mRNA,IL-6 mRNA were increased in the HG group(P<0.05);compared with the HG group,the expression levels of TNF-αmRNA,IL-1 mRNA and IL-6 mRNA in the HG+Res group decreased(P<0.05).Conclusion Res may delay the damage of PRE cells by high glucose environment through anti-oxidation,inhibition of proliferation,and anti-inflammatory.
作者
李青春
李岱
周跃
周燕红
LI Qingchun;LI Dai;ZHOU Yue;ZHOU Yanhong(Department of Ophthalmology,Xianning Aier Eye Hospital,Xianning,Hubei 437000,China;Medical School of Ophthalmology and Otorhinolaryngology,Hubei University of Science and Technology,Xianning,Hubei 437100,China)
出处
《重庆医学》
CAS
2020年第18期3002-3007,共6页
Chongqing medicine
基金
2016年度爱尔眼科医院集团科研基金自由申报立项项目(AF151D02)。
关键词
白藜芦醇
视网膜色素上皮
氧化性应激
炎症
resveratrol
retinal pigment epithelium
oxidative stress
inflammation
