甘草查尔酮A对过氧化氢致视网膜神经节细胞损伤的保护作用  

Protective effect of licochalcone A on the injure of retinal ganglion cell induced by hydrogen peroxide in vitro

作　　者：王亮[1] 田莹[1] 程钰[1] 赵俊宏[1] 曹燕[1] 郭建强[1] WANG Liang;TIAN Ying;CHENG Yu;ZHAO Jun-Hong;CAO Yan;GUO Jian-Qiang(Departmentof Ophthalmology,Xi’an No.1 Hospital,Xi’an 710002,Shaanxi Province,China)

机构地区：[1]西安市第一医院眼科,陕西省西安市710002

出　　处：《眼科新进展》2018年第11期1037-1040,共4页Recent Advances in Ophthalmology

摘　　要：目的研究甘草查尔酮A (licochalcone A,LCA)对过氧化氢(H_2O_2)致视网膜神经节细胞(retinal ganglion cells,RGC)损伤的保护作用,为LCA在青光眼治疗方面的应用提供初步的实验依据。方法体外培养RGC-5细胞,随机分为对照组(正常培养基培养)、H_2O_2组(培养基中加入200μmol·L^(-1)H_2O_2)、LCA组(培养基中除加入200μmol·L^(-1) H_2O_2外,还分别加入10μmol·L^(-1)、20μmol·L^(-1)、40μmol·L^(-1)的LCA溶液),CCK-8法检测各组细胞增殖活性,流式细胞术检测各组细胞凋亡情况,Western blotting检测各组细胞中Bax、Bcl-2和Cleaved Caspase-3蛋白表达水平,ELISA法检测各组细胞中丙二醛含量和超氧化物歧化酶活性。结果 CCK-8法检测结果显示,10μmol·L^(-1)、20μmol·L^(-1)和40μmol·L^(-1) LCA组的细胞存活率分别为(63.7±5.4)%、(78. 8±6. 3)%和(72. 3±6.9)%,与H_2O_2组的(54. 3±3. 9)%相比,20μmol·L^(-1) LCA组增殖最显著(P<0.05)。流式细胞术检测结果显示,10μmol·L-1、20μmol·L^(-1)和40μmol·L^(-1) LCA组细胞凋亡率分别为(21.1±1.9)%、(12.3±1.3)%和(14. 8±2.0)%,与H_2O_2组的(29. 3±2.0)%比较,差异均有统计学意义(均为P <0.05)。Western blotting检测结果显示,与H_2O_2组比较,不同浓度LCA组Bcl-2蛋白的表达均增加,Bax和Cleaved Caspase-3蛋白的表达均降低,差异均有统计学意义(均为P<0.05)。ELISA法检测结果显示,与H_2O_2组比较,不同浓度LCA组RGC-5的丙二醛含量均降低,但超氧化物歧化酶活性均显著增加(均为P<0.05)。结论LCA能缓解H_2O_2诱导的RGC损伤,其作用机制可能与抑制细胞凋亡和氧化应激有关。Objective To investigate the effect of Licochalcone A(LCA)on cell injure of retinal ganglion cell(RGC)induced by hydrogen peroxide(H 2O 2)in vitro,and to provide preliminary experimental basis for the treatment of glaucoma by LCA.Methods RGC-5 cells were cultured in vitro,and then were randomly divided into control group(normal medium),H 2O 2 group with 200μmol·L-1 H 2O 2 in the normal medium,and LCA group with 200μmol·L-1 H 2O 2 and 10μmol·L-1,20μmol·L-1,or 40μmol·L-1 LCA in normal medium,respectively.Cell proliferation of RGC-5 cells was detected by CCK-8 assay.Cell apoptosis was examined with flow cytometry methods.The protein expression level of Bax,Bcl-2 and Cleaved Caspase-3 was detected by Western blot.The content of malondialdehyde and the activity of superoxide dismutase were detected by ELISA assay.Results The results of CCK-8 assay showed that cell viability of RGC-5 cells in the 10μmol·L-1,20μmol·L-1,and 40μmol·L-1 LCA group was(63.7±5.4)%,(78.8±6.3)%,and(72.3±6.9)%,respectively.Compared with the H 2O 2 group[(54.3±3.9)%],20μmol·L-1 LCA had the greatest effect(P<0.05).The results of flow cytometry methods indicated that the rate of cell apoptosis in the 10μmol·L-1,20μmol·L-1,and 40μmol·L-1 LCA treatment group was(21.1±1.9)%,(12.3±1.3)%,and(14.8±2.0)%,respectively.The difference of all LCA group was statistically significant(all P<0.05)compared with the H 2O 2 group[(29.3±2.0)%].The results of Western blot showed that the Bcl-2 protein expression in group with different concentrations of LCA was up-regualted,and the protein expression level of Bax and Cleaved Caspase 3 was down-regualted,all with significant difference(all P<0.05)when compared with H 2O 2 group.The results of ELISA assay showed that the content of malondialdehyde in group with different concentrations of LCA was diminished and the activity of superoxide dismutase was increased,all with significant difference(all P<0.05)when compared with H 2O 2 group.Conclusion LCA relieves the cell injure of RGC induced

关 键 词：甘草查尔酮A 过氧化氢 视网膜神经节细胞 细胞凋亡 

分 类 号：R775[医药卫生—眼科]