黄芩苷对高糖诱导的人晶状体上皮细胞凋亡和氧化应激的影响及其可能机制  

Effects of baicalin on apoptosis and oxidative stress of human lens epithelial cells induced by high glucose and its possible mechanisms

作　　者：杨涛 柴盼盼[2] 何启红 潘家钰 彭正虹 康刚劲 YANG Tao;CHAI Panpan;HE Qihong;PAN Jiayu;PENG Zhenghong;KANG Gangjin(Department of Ophthalmology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Department of Ophthalmology,Luzhou Hospital of Traditional Chinese Medicine,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院眼科,四川省泸州市646000 [2]泸州市中医医院眼科,四川省泸州市646000

出　　处：《眼科新进展》2021年第5期422-427,共6页Recent Advances in Ophthalmology

基　　金：西南医科大学-泸州市中医医院基地项目(编号:2019-LH007)。

摘　　要：目的探讨黄芩苷对高糖诱导的人晶状体上皮细胞凋亡和氧化应激的影响,并探讨其可能机制。方法以人晶状体上皮细胞系HLE-B3为研究对象,用不同浓度黄芩苷处理后,采用CCK-8法检测细胞活力以筛选黄芩苷最佳作用浓度用于后续实验。将细胞随机分为正常组、高糖组、黄芩苷组,采用CCK-8法检测各组细胞活力,流式细胞术检测各组细胞凋亡率,RT-PCR检测Kelch样ECH相关蛋白1(Keap1)、NQO1、核因子红细胞2相关因子2(Nrf2)、HO-1、Bax、Bcl-2的mRNA相对表达量,Western blot检测Keap1、Nrf2、NQO1、HO-1、Bax、Bcl-2的蛋白表达水平,酶标法检测各组细胞超氧化物歧化酶(SOD)活力和丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)含量。结果CCK-8法检测结果显示,5μmol·L^(-1)黄芩苷可以显著增强HLE-B3细胞活力,随着黄芩苷浓度升高,细胞活力下降,故以5μmol·L^(-1)黄芩苷进行后续实验。与正常组HLE\|B3细胞活力[(100.00±0.00)%]相比,高糖组细胞活力[(73.52±1.71)%]显著降低(P<0.01);与高糖组相比,黄芩苷组细胞活力[(92.36±3.61)%]显著升高(P<0.01)。流式细胞术检测结果显示,与正常组HLE-B3细胞凋亡率[(7.93±0.22)%]相比,高糖组细胞凋亡率[(57.12±2.63)%]显著升高(P<0.01);与高糖组相比,黄芩苷组细胞凋亡率[(42.09±1.04)%]显著降低(P<0.01)。RT\|PCR检测结果显示,与正常组相比,高糖组HLE\|B3细胞中Nrf2、HO-1、NQO1、Bcl-2 mRNA相对表达量均显著降低,Keap1、Bax mRNA相对表达量均显著升高(均为P<0.05);与高糖组相比,黄芩苷组HLE\|B3细胞中Nrf2、NQO1、HO-1、Bcl-2 mRNA相对表达量均显著升高,Keap1、Bax mRNA相对表达量均显著降低(均为P<0.05)。Western blot检测结果显示,高糖组HLE\|B3细胞中Keap1、Bax蛋白相对表达量均较正常组显著升高,黄芩苷组Keap1、Bax蛋白相对表达量均较高糖组显著降低(均为P<0.05);高糖组HLE\|B3细胞中Nrf2、HO-1、NQO1、Bcl-2蛋白相对表达量均较正Objective To explore effect of baicalin to the apoptosis and oxidative stress of human lens epithelial cells(HLE-B3)induced by high glucose and its possible mechanisms.Methods CCK-8 was used to detect the proliferation rate of HLE-B3 which was cultured in vitro with a certain concentration of baicalin.According to the proliferation rate of HLE-B3 detected by CCK-8,the optimal concentration of baicalin was selected for follow-up experiments.HLE-B3 cells were divided into three groups randomly including control group,high glucose group and baicalin group.CCK-8 and flow cytometry were used to detect the proliferation and apoptosis rate of every group,respectively.RT-PCR was used to detect the expression of mRNAs of Keap1,NQO1,Nrf2,HO-1,Bax and Bcl-2.Western blot was used to detect the expression levels of proteins of Keap1,NQO1,Nrf2,HO-1,Bax and Bcl-2.Enzyme labeling method was used to detect the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)in each group.Results CCK-8 results showed the proliferation of HLE-B3 cultured in 5μmol·L^(-1)baicalin was higher than other group,so we chosed it for follow-up experiments.The cell activity of HLE-B3 in the high glucose group was(73.52±1.71)%,which was significantly lower than that in the control group[(100.00±0.00)%],and the baicalin group[(92.36±3.61)%]was higher than the high glucose group(both P<0.01).Flow cytometry showed that the apoptosis rate of high glucose group[(57.12±2.63)%]was higher than that in control group[(7.93±0.22)%],and the baicalin group[(42.09±1.04)%]was lower than the high glucose group(both P<0.01).RT-PCR showed that,when compared with the control group,the expression levels of mRNAs of Nrf2,HO-1,NQO1 and Bcl-2 of high glucose group decreased,while the expression levels of Keap1 and Bax increased(all P<0.05).The expression levels of mRNAs of Nrf2,NQO1,HO-1 and Bcl-2 in baicalin group was higher than those in the high glucose group,while the expression levels of Keap1 and Bax mRNAs

关 键 词：高糖 晶状体上皮细胞 细胞凋亡 黄芩苷 氧化应激 

分 类 号：R776.1[医药卫生—眼科]