异烟肼对人肝细胞CYP1A1启动子区甲基化的影响

发布时间：2018-06-27 16:49

本文选题：异烟肼 + 药物性肝损伤　；参考：《山东医药》2017年26期

【摘要】：目的观察异烟肼对人肝细胞细胞色素P4501A1(CYP1A1)启动子区CpG岛甲基化的影响,探讨异烟肼致肝细胞损伤的机制。方法将肝细胞分为观察组和对照组,观察组分别加入200、400、800μg/mL异烟肼,对照组加入同体积含DMSO的培养液。采用乳酸脱氢酶(LDH)检测试剂盒检测细胞培养上清液中的LDH水平来观察细胞损伤情况,实时荧光定量PCR方法检测细胞CYP1A1、DNA甲基转移酶1(DNMT1)、DNMT3a、DNMT3b mRNA的相对表达量,酶联免疫吸附法检测细胞DNMT1、DNMT3a、DNMT3b蛋白水平,采用亚硫酸盐修饰后测序法检测CYP1A1启动子区甲基化状态。结果观察组细胞培养上清液中的LDH水平均高于对照组,800μg/mL异烟肼组最高(P均0.05)。与对照组比较,观察组随异烟肼浓度升高,CYP1A1 mRNA表达呈先升高后降低趋势,差异均有统计学意义(P均0.05);DNMT1、DNMT3a、DNMT3b mRNA和蛋白表达均随异烟肼浓度升高而升高,800μg/mL异烟肼组高于其他组(P均0.05)。细胞CYP1A1基因启动子区甲基化率为82.9%,高于对照组的79.6%(P0.05)。结论异烟肼可导致肝细胞CYP1A1基因启动子区高甲基化,进而引起肝细胞损伤。
[Abstract]:Objective to observe the effect of isoniazid on the methylation of CpG island in the promoter region of human hepatocyte cytochrome P450 1A1 (CYP1A1) and to explore the mechanism of isoniazid induced hepatocyte injury. Methods Hepatocytes were divided into observation group and control group. 200400800 渭 g / mL isoniazid was added into the observation group and the same volume of DMSO medium was added to the control group. Lactate dehydrogenase (LDH) assay kit was used to detect the level of LDH in the supernatant of cell culture to observe the cell damage. The relative expression of DNMT3aI DNMT3b mRNA in CYP1A1DNA methyltransferase 1 (DNMT1) was detected by real-time fluorescence quantitative PCR. Enzyme linked immunosorbent assay (Elisa) was used to detect the level of DNMT1DNMT3b protein, and the methylation status of CYP1A1 promoter was detected by sulfite modified DNA sequencing. Results the LDH level in the supernatant of cell culture in the observation group was higher than that in the control group (800 渭 g / mL isoniazid) (P 0.05). Compared with the control group, the mRNA expression of CYP1A1 increased first and then decreased with the increase of isoniazid concentration in the observation group, and the difference was statistically significant (P 0.05). The mRNA and protein expressions of DNMT1 and DNMT3ahe DNMT3b increased with the increase of isoniazid concentration in the isoniazid group (800 渭 g / mL) compared with the other groups (P 0.05). The methylation rate of CYP1A1 promoter was 82.9%, which was higher than that of control group (79.6%) (P0.05). Conclusion Isoniazid can induce hypermethylation of CYP1A1 gene promoter in hepatocytes and induce hepatocyte damage.
【作者单位】：华北理工大学公共卫生学院;
【基金】：国家自然科学基金资助项目(81041096)
【分类号】：R965

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