藏红花酸预处理对高糖诱导人视网膜色素上皮细胞损伤的影响  

Effect of crocetin pretreatment on high glucose induced injury of human retinal pigment epithelial cells

作　　者：王爽[1] 王含伊 刘娟[1] WANG Shuang;WANG Han-yi;LIU Juan(Department of Ophthalmology and Otorhinolaryngology Operation Room,The First Affiliated Hospital of Harbin Medical University,Harbin J50001,China)

机构地区：[1]哈尔滨医科大学附属第一医院五官科手术室,黑龙江哈尔滨150001

出　　处：《哈尔滨医科大学学报》2022年第1期8-11,共4页Journal of Harbin Medical University

摘　　要：目的 探讨藏红花酸对高糖诱导人视网膜色素上皮(human retinal pigment epithelial, HRPE)细胞氧化应激损伤的保护作用。方法 将HRPE细胞分为对照组:正常DMEM培养基(含5.5 mmol/L D-葡萄糖)进行培养;模型组:高糖(含50 mmol/L D-葡萄糖)DMEM培养液培养;藏红花酸低、高浓度组:用藏红花酸(1μmol/L、10μmol/L)处理细胞24 h后,再将细胞培养于含50 mmol/L D-葡萄糖的DMEM培养基内。CCK-8法观察细胞增殖率,ELISA法检测谷氨酸、细胞超氧化物歧化酶(superoxide dismutase, SOD)及丙二醛(malondialdehyde, MDA)变化,2′,7′-二氯荧光素染色检测细胞氧化应激水平,Western blot分析核因子E2相关因子2(nuclear factor erythriod 2-related factor 2,Nrf-2)及抗氧化酶血红素氧合酶-1(heme oxygenase-1,HO-1)的变化。结果 CCK-8结果显示,藏红花酸处理后HRPE细胞增殖率升高,与模型组比较差异具有统计学意义(P<0.05)。ELISA结果显示,藏红花酸干预后谷氨酸、MDA含量均下降,与模型组比较差异有统计学意义(P<0.05);SOD表达水平升高,与模型组比较差异有统计学意义(P<0.05)。2′,7′-二氯荧光素染色结果显示,藏红花酸逆转了高糖介导的HRPE细胞中ROS的升高。Western blot印迹法结果显示,藏红花酸干预后Nrf-2及HO-1蛋白表达上调,与模型病组相比差异均有统计学意义(P<0.05)。结论 藏红花酸可以保护HRPE细胞免受高糖引起的氧化应激损伤。Objective To investigate the protective effect of crocetin on high glucose induced oxidative stress injury in human retinal pigment epithelial(HRPE) cells. Methods HRPE cells were divided into control group: normal DMEM medium(containing 5.5 mmol/L D-glucose) for culture;model group: high glucose(containing 50 mmol/L D-glucose) DMEM medium for culture;crocetin low and high concentration group: crocetin(1 μmol/L and 10 μmol/L) for 24 h, then treated as model group. Cell proliferation rate was observed by CCK-8 assay, the changes of glutamic acid, superoxide dismutase(SOD) and malondialdehyde(MDA) were detected by ELISA, the level of oxidative stress was detected by 2′,7′-Dichlorofluorescein staining, changes of nuclear factor erythriod 2-related factor 2(Nrf-2) and heme oxygenase-1(HO-1) were analyzed by Western blot. Results CCK-8 results showed that the proliferation rate of HRPE cells increased after saffron treatment, the difference was statistically significant compared with the model group(P<0.05). The results of ELISA showed that the contents of glutamate and MDA decreased after saffron intervention, which was statistically significant compared with model group(P<0.05);The expression level of SOD increased, which was statistically significant compared with model group(P<0.05). The results of 2′,7′-Dichlorofluorescein staining showed that saffron reversed the increase of ROS in HRPE cells mediated by high glucose. The results of Western blot showed that the expression of Nrf-2 and HO-1 protein were up-regulated after saffron intervention, which was statistically significant compared with model group(P<0.05). Conclusion Crocetin could protect HRPE cells from oxidative stress injury induced by high glucose.

关 键 词：藏红花酸 高糖 视网膜色素上皮细胞 氧化应激 

分 类 号：R774.1[医药卫生—眼科]