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马玲玲, 范秋玉, 肖德强, 刘丹丹, 邓祥发, 贾亮, 黄东萍, 鲁力. PC-B2对AFB1致肝细胞DNA损伤及修复影响[J]. 中国公共卫生, 2016, 32(7): 925-927. DOI: 10.11847/zgggws2016-32-07-12
引用本文: 马玲玲, 范秋玉, 肖德强, 刘丹丹, 邓祥发, 贾亮, 黄东萍, 鲁力. PC-B2对AFB1致肝细胞DNA损伤及修复影响[J]. 中国公共卫生, 2016, 32(7): 925-927. DOI: 10.11847/zgggws2016-32-07-12
MA Ling-ling, FAN Qiu-yu, XIAO De-qiang.et al, . Influence of PC-B2 on oxidative DNA damage induced by AFB1 and repair gene expression in hepatocytes[J]. Chinese Journal of Public Health, 2016, 32(7): 925-927. DOI: 10.11847/zgggws2016-32-07-12
Citation: MA Ling-ling, FAN Qiu-yu, XIAO De-qiang.et al, . Influence of PC-B2 on oxidative DNA damage induced by AFB1 and repair gene expression in hepatocytes[J]. Chinese Journal of Public Health, 2016, 32(7): 925-927. DOI: 10.11847/zgggws2016-32-07-12

PC-B2对AFB1致肝细胞DNA损伤及修复影响

Influence of PC-B2 on oxidative DNA damage induced by AFB1 and repair gene expression in hepatocytes

  • 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、30 μg/mL)、AFB1染毒组(10、20、30、40 μg/mL)和PC-B2干预组(3、10、30 μg/mL PC-B2+30 μg/mL 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-OHdG)含量及细胞hOGG1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,30 μg/mL AFB1组细胞活力(69.9±2.46)%明显降低(P < 0.05),细胞上清中8-OHdG含量(2.779±0.089)ng/mL明显升高(P < 0.05);与30 μg/mL AFB1组比较,3、10、30 μg/mL 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-OHdG 含量分别为(2.550±0.078)、(2.376±0.109)、(1.873±0.065)ng/mL明显降低(P < 0.05);与溶剂对照组比较,30 μg/mL AFB1组L-02细胞hOGG1基因表达减少(P < 0.05);与30 μg/mL AFB1组比较,PC-B2干预组L-02细胞hOGG1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hOGG1表达有关。

     

    Abstract: 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 supernatant and cellular human 8-oxoguanine DNA glycosylase-1 (hOGG1) expression were assayed with 3-(4, 5-dimethylthiazolyl)-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 decreased and the content of 8-OhdG (2.779±0.089 ng/mL)was significantly increased in 30 μg/mL AFB1 exposure group (P < 0.05).Compared with 30 μg/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.

     

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