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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 effects of proanthocyanidin B2 (PC-B2) on DNA damage and repair gene expression of human embryonic hepatocytes (L-02) induced by aflatoxin B1 (AFB1). Methods L-02 cells with good logarithmic growth phase were randomly divided into blank control group, solvent control group, PC-B2 treatment group (3,10 and 30μg / mL), AFB1 exposure group (10,20,30, 40μg / m L) and PC-B2 intervention group (3, 10, 30μg / m L PC-B2 + 30μg / m L AFB1). The cell proliferation was measured by the method of thiazolyl blue, enzyme-linked immunosorbent assay and real- Vitality, 8-OHdG (8-OHdG) content and h OGG1 gene expression level in the cell supernatant. Results AFB1 significantly inhibited the proliferation of L-02 cells (P <0.05) in a dose-response manner. Compared with the solvent control group, the viability of AFB1 cells in 30 μg / mL AFB1 group was significantly lower than that in the control group (69.9 ± 2.46% 0.05). The content of 8-OHd G in the supernatant was significantly higher than that in the AFB1 group ([(2.779 ± 0.089) ng / m L] (P <0.05) -B2 intervention group (70.6 ± 2.67%, (69.7 ± 1.94)%, (82.4 ± 1.58)%] (P <0.05), and the content of 8-OHd G in the cell supernatant (2.550 ± 0.078), (2.376 ± 0.109), (1.873 ± 0.065) ng / m L] (P <0.05). Compared with the solvent control group, h OGG1 gene in 30μg / mL AFB1 group (P <0.05). Compared with 30μg / ml AFB1 group, the expression of hGG1 in L-02 cells was significantly increased in PC-B2 intervention group. Conclusion PC-B2 can increase the proliferation of hepatocytes and inhibit the DNA damage induced by AFB1, which may be related to the regulation of h OGG1 gene expression.