PC-B2对AFB_1致肝细胞DNA损伤及修复影响  

Influence of PC-B2 on oxidative DNA damage induced by AFB_1 and repair gene expression in hepatocytes

作　　者：马玲玲[1] 范秋玉[1] 肖德强[1] 刘丹丹[2] 邓祥发[3] 贾亮[4] 黄东萍[1] 鲁力[1] 

机构地区：[1]广西医科大学公共卫生学院,广西南宁530021 [2]广西医科大学护理学院 [3]广西医科大学基础医学院 [4]广西壮族自治区疾病预防控制中心

出　　处：《中国公共卫生》2016年第7期925-927,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(31360383)

摘　　要：目的探讨原花青素B2(PC-B2)对黄曲霉毒素B1(AFB1)所致人胚胎肝细胞(L-02)DNA损伤及修复基因表达影响。方法取对数期生长良好的L-02细胞随机分为空白对照组、溶剂对照组、PC-B2处理组(3、10、30μg/m L)、AFB1染毒组(10、20、30、40μg/m L)和PC-B2干预组(3、10、30μg/m L PC-B2+30μg/m L AFB1),利用噻唑蓝法、酶联免疫吸附法和荧光定量PCR技术分别测定细胞增殖活力、细胞上清液8-羟基脱氧鸟苷(8-OHd G)含量及细胞h OGG1基因表达水平。结果 AFB1可明显抑制L-02细胞增殖活力(P<0.05),呈剂量-效应关系;与溶剂对照组比较,30μg/m L AFB1组细胞活力[(69.9±2.46)%]明显降低(P<0.05),细胞上清中8-OHd G含量[(2.779±0.089)ng/m L]明显升高(P<0.05);与30μg/m L AFB1组比较,3、10、30μg/m L PC-B2干预组细胞活力[分别为(70.6±2.67)%、(69.7±1.94)%、(82.4±1.58)%]明显升高(P<0.05),细胞上清中8-OHd G含量[分别为(2.550±0.078)、(2.376±0.109)、(1.873±0.065)ng/m L]明显降低(P<0.05);与溶剂对照组比较,30μg/m L AFB1组L-02细胞h OGG1基因表达减少(P<0.05);与30μg/m L AFB1组比较,PC-B2干预组L-02细胞h OGG1表达明显升高。结论 PC-B2可提高肝细胞增殖活力,抑制AFB1所致肝细胞DNA损伤,其机制可能与调控修复基因h OGG1表达有关。Objective To investigate the effect of procyanidin B2 （PC-B2） on oxidative DNA damage induced by aflaoxin B1 （ AFB1 ） and repair gene expression in hepatocytes. Methods Well grown L-02 cells in logarithmic phase were divided into control group, dimethyl sulfoxide （DMSO） group, AFB1 exposure groups （ 10,20,30, and 40 μg/ mL）, PC-B2 exposure groups （3,10, and 30 μg/mL）, and PC-B2 intervention groups （3,10, and 30 μg/mL PC-B2 ± 30 μg/mL AFB1 ）. Cell proliferation and the levels of 8-hydroxy-2＇-deoxyguanosine （8-OHdG） in cell culture superna- tant and cellular human 8-oxoguanine DNA glycosylase-1 （hOGG1） expression were assayed with 3-（4,5-dimethylthia- zolyl）-2,5-diphenyltetrazolium bromide （MTT）, enzyme-linked immunosorbent assay （ELISA）, and quantitative PCR （qPCR） method. Results The cell proliferation in AFB1 exposure groups was significantly inhibited, with a dose-effect relationship （P 〈 0. 05 ）. Compared with that of DMSO group,the cell proliferation （69. 9 ± 2. 46% ） was obviously de- creased and the content of 8-OhdG （2. 779 ±0. 089 ng/mL）was significantly increased in 30 p.g/mL AFB1 exposure group （ P 〈 0. 05 ）. Compared with 30 iμg/mL AFB1 exposure group, the cell proliferations （ 70. 6 ±2. 67%, 69. 7 ±1.94%, and 82.4 ± 1.58% ） were increased and the contents of 8-OHdG （ 2. 550 ±0. 078,2. 376 ± 0. 109, and 1. 873 ±0. 065 ng/mL） were significantly decreased in the three PC-B2 treatment groups （ all P 〈 0. 05 ） ; the expression of hOGG1 decreased in the 30 μg/mL AFB1 treatment group ,but it obviously increased after the treatment of PC-B2 （P 〈 0. 05）. Conclusion PC-B2 has the effect of increasing the proliferation of hepatocyte and inhibiting oxidative DNA damage induced by AFB1. The antioxidant activity may related to hOGG1 gene expression levels.

关 键 词：原花青素-B2(PC-B2) 黄曲霉毒素B1(AFB1) 人胚胎肝细胞(L-02) 8-羟基脱氧鸟苷(8-OHdG) 人8-羟基鸟嘌呤DNA糖苷酶(hOGG1) 

分 类 号：R735.7[医药卫生—肿瘤]