黄连素通过Nrf2/HO-1通路对过氧化氢诱导的人晶状体上皮细胞-B3损伤的影响  

Effects of berberine on human lens epithelial cell-B3 injury induced by hydrogen peroxide through Nrf2/HO-1 pathway

作　　者：梁圆圆 杨彬 于亚东 胡馨 赵博 LIANG Yuanyuan;YANG Bin;YU Yadong;HU Xin;ZHAO Bo(Department of Ophthalmology,Huaihe Hospital,Henan University,Kaifeng,Henan 475000)

机构地区：[1]河南大学淮河医院眼科,河南开封475000

出　　处：《郑州大学学报(医学版)》2025年第2期153-158,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20230443)。

摘　　要：目的:探讨黄连素通过核因子E2相关因子2/血红素加氧酶-1(Nrf2/HO-1)信号通路对过氧化氢造成的人晶状体上皮细胞-B3(HLE-B3)损伤的影响。方法:取对数生长期HLE-B3细胞分为对照组,氧化损伤组,低、中和高剂量黄连素组,各剂量黄连素组细胞分别用含50、100和200μmol/L黄连素的培养基100μL培养24 h,之后换用含300μmol/L过氧化氢的培养基100μL培养24 h,氧化损伤组用仅含300μmol/L过氧化氢的培养基、对照组仅更换新鲜培养基培养24 h。MTT法检测细胞增殖活性,Annexin V-FITC/PI双染法检测细胞凋亡率,ELISA法检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平,免疫荧光法检测活性氧(ROS)水平,qRT-PCR法和Western blot法检测Nrf2、HO-1、诱导型一氧化氮合酶(iNOS)、环氧合酶-2(COX-2)mRNA和蛋白相对表达量。结果:与对照组比较,氧化损伤组细胞增殖活性,SOD和GSH-Px活性,Nrf2和HO-1 mRNA和蛋白相对表达量降低;细胞凋亡率,MDA含量,ROS水平,iNOS、COX-2 mRNA和蛋白相对表达量升高(P<0.05)。与氧化损伤组比较,低、中和高剂量黄连素组细胞增殖活性,SOD和GSH-Px活性,Nrf2和HO-1 mRNA和蛋白相对表达量升高;细胞凋亡率,MDA含量,ROS水平,iNOS、COX-2 mRNA和蛋白相对表达量降低,且高剂量黄连素作用最强(P<0.05)。结论:黄连素可抑制过氧化氢造成的HLE-B3细胞凋亡,促进增殖活性,降低氧化应激反应,其可能是通过激活Nrf2/HO-1通路发挥作用。Aim:To investigate the effects of berberine on human lens epithelial cell-B3(HLE-B3)injury caused by hydrogen peroxide through nuclear factor E2 related factor 2/heme oxygenase-1(Nrf2/HO-1)pathway.Methods:HLE-B3 cells in the log phase were divided into control group,oxidative injury group,low,medium and high dose berberine group.The cells in berberine groups were cultured with 100μL medium containing 50,100 and 200μmol/L berberine,respectively for 24 hours,and then cultured with 100μL medium containing 300μmol/L hydrogen peroxide for 24 hours.Cells in the oxidative injury group were only cultured with 300μmol/L hydrogen peroxide medium for 24 hours,while cells in control group were only cultured with fresh medium.Cell proliferation activity was detected by MTT assay,cell apoptosis rate was detected by staining with Annexin V-FITC/PI,malondialdehyde(MDA)content,superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)activities were detected by ELISA,and reactive oxygen species(ROS)level was detected by immunofluorescence assay.The mRNA and protein expression levels of Nrf2,HO-1,inducible nitric oxide synthase(iNOS)and cyclooxygenase 2(COX-2)were detected by qRT-PCR and Western blot.Results:Compared with control group,cell proliferation activity,SOD and GSH-Px activity,Nrf2 and HO-1 mRNA and protein expression levels decreased in the oxidative injury group,while cell apoptosis rate,MDA content,ROS level,iNOS and COX-2 mRNA and protein expression levels increased(P<0.05).Compared with the oxidative injury group,cell proliferation activity,SOD and GSH-Px activities,Nrf2 and HO-1 mRNA and protein expressions increased in low,medium and high dose berberine groups,while cell apoptosis rate,MDA content,ROS level,iNOS and COX-2 mRNA and protein expression levels decreased(P<0.05).The high dose berberine had the strongest effect.Conclusion:Berberine could inhibit the apoptosis of HLE-B3 cells caused by hydrogen peroxide,promote proliferation activity and reduce oxidative stress response,which may be associated with

关 键 词：过氧化氢 人晶状体上皮细胞-B3 黄连素 Nrf2/HO-1通路 

分 类 号：R776.1[医药卫生—眼科]