微胶囊化儿茶素拮抗H_2O_2诱导的大鼠GMCs DNA损伤  

Effect of catechin microcapsulation on the repair of DNA damage in glumreular mesangial cells induced by H_2O_2

作　　者：何小解[1] 易著文[1] 莫双红[1] 党西强[1] 卢向阳[2] 田云[2] 白海涛[1] 

机构地区：[1]中南大学湘雅二医院小儿肾脏病研究室 [2]湖南农业大学,长沙410128

出　　处：《中南大学学报（医学版）》2007年第1期82-87,共6页Journal of Central South University ：Medical Science

基　　金：湖南省自然科学基金(02JJXY3020);湖南省教育厅重点项目基金(02A015)~~

摘　　要：目的:研究微胶囊化儿茶素对过氧化氢(H2O2)诱导的肾小球系膜细胞(glumreular mesang-ial cells,GMCs)DNA损伤修复作用及可能机制。方法:按H2O2终浓度分为0(对照组)、50μmol/L组、100μmol/L组、150μmol/L组、200μmol/L组、250μmol/L组,每个处理组又分6,12,24h3个小组,筛选过氧化氢对GMCsDNA损伤研究的最适剂量与最佳时点。然后将培养细胞分生理盐水对照组、H2O2组、不同浓度儿茶素组(按终浓度又分为10.0,15.0,20.0mg/L3个组)与不同浓度微胶囊化儿茶素组(按胶囊中儿茶素含量终浓度又分为10.0,15.0,20.0mg/L3个组),共8组进行培养。利用生物化学法检测各组大鼠GMCs培养上清中羟自由基(hydroxy radical,.OH-)、丙二醛(malonydialdehyde,MDA)与总超氧化物歧化酶(total superoxide dismutase,tSOD)含量,利用单细胞凝胶电泳技术检测大鼠GMCsDNA损伤修复。结果:(1)6h时,100μmoL/L过氧化氢剂量组即可引起GMCs DNA损伤,150μmoL/L组则可检测到DNA出现明显损伤;(2)微胶囊化儿茶素组GMCs慧星全长在不同剂量时与儿茶素组差异无统计学意义,微胶囊化儿茶素组GMCs慧星尾长与尾部荧光强度在不同剂量时均低于儿茶素组,差异有统计学意义(均P<0.05,0.01);(3)10.0mg/L时,儿茶素组与微胶囊化儿茶素两组之间相比,两组.OH-,MDA与tSOD含量差异无统计学意义;15.0mg/L与20.0mg/L剂量时,微胶囊化儿茶素组.OH-和MDA含量较儿茶素组降低,tSOD含量较儿茶素组增高,差异有统计学意义(均P<0.05)。结论:微胶囊化儿茶素可能是通过提高其抗氧化能力而提高其对DNA损伤保护及损伤修复能力。Objective To explore the effect and possible mechanism of catechin microcapsulation on the repair of DNA damage in glumreular mesangial cells （GMCs） induced by H2O2. Methods According to H2O2 concentration, the experiment GMCs were divided into 6 groups： a control group, 50 μmol/L group, 100 μmol/L group, 150 μmol/L group, 200 μmol/L group and 250 μmol/L group, Each group was sub-divided into 3 groups： 6 h group, 12 h group and 24 h group,in order to determining the optimum dose and the best time of detecting the DNA damage in GMCs. The cultured cells were divided into 8 groups as follows： the NS control group, the H2 02 group, the catechin groups （ the final concentrations were 10.0,15.0, and 20.0 mg/L respectively ） and the various catechin microcapsulation groups （ the final concentrations were 10.0, 15.0, and 20.0 mg/L respectively ）. At the end of the experiment, hydroxy radical （ OH ）, malonydialdehyde （ MDA ） and total superoxide dismutase （tSOD） concentration of supernadant in GMCs were determined by biochemistry assay, the repair of DNA damage in GMCs were detected by single cell gel electrophoresis assay. Results （ 1 ） At 6^th h, H2O2 of 100 μmoL/L could cause the DNA damage of GMCs, and H2O2 of 150 μmol/L could result in DNA damage significantly. （2） No difference was found in the comet span of GMCs DNA in the catechin group and catechin microcapsulation group of different concentrations, while the DNA comet tail-long in the catechin microcapsulation group was shorter than that of the catechin group （ all P 〈 0.05 ） , and the fluorescence intensity of tail in the catechin microcapsulation group was lower than that of the catechin group （ all P 〈 0.01 ）. （ 3 ） When the concentration of catechin was 10.0 mg/L, no statistical significance was obtained in the concentration of · OH^-, MDA and tSOD between the catechin microcapsulation group and the catechin group; while ·OH^- and MDA concentrations were lower, and the tSOD was higher in the cate

关 键 词：儿茶素 微胶囊 单细胞凝胶电泳 DNA 肾小球系膜细胞 

分 类 号：R726.9[医药卫生—儿科]