砷对Keap1基因沉默的人皮肤角质细胞砷甲基化代谢酶表达的影响研究

发布时间：2018-02-22 07:58

  本文关键词： Nrf Keap 砷代谢酶　出处：《现代预防医学》2017年15期 　论文类型：期刊论文

【摘要】：目的砷对Keap1基因沉默的人皮肤角质(Ha Ca T)细胞砷甲基化代谢酶表达的影响。方法实验共五组,A组为正常Ha Ca T细胞,B、C、D、E组为模型细胞。A、B组不给砷,C、D、E组给予不同浓度的砷。五组细胞在8h、24h、48h、72h,用实时荧光定量PCR(q PCR)检测核转录因子红胞系-2p45(NF-E2)相关因子-2(Nrf2)、KELCH样环氧氯丙烷相关蛋白1(Keap1)、三价砷甲基转移酶(As3MT)、N-6-腺嘌呤DNA甲基转移酶1(N6AMT1)的基因表达;酶联免疫吸附实验(ELISA)检测细胞内同型半胱氨酸(HCY)、S-腺苷甲硫氨酸(SAM)、As3MT、N6AMT1的蛋白浓度。结果(1)QPCR结果发现:与A组比,抑制Keap1基因使细胞中N6AMT1基因表达在8h、24h、48h表达上调(P0.05)。与B组比:砷浓度2.9μmol/L,8h、72h N6AMT1、24h As3MT基因表达上调(P0.017),8h、72h As3MT、24h N6AMT1基因表达下调(P0.017);砷浓度5.8μmol/L,8h N6AMT1、48h As3MT基因表达上调(P0.017),8h As3MT、48h N6AMT1基因表达下调(P0.017)。砷浓度29μmol/L时,24h N6AMT1基因表达下调(P0.017),As3MT基因表达上调(P0.017)。(2)ELISA结果发现:与B组比,砷浓度≤5.8μmol/L SAM、HCY蛋白表达上调(P0.0125);砷浓度29μmol/L抑制SAM、HCY蛋白的表达下调(P0.0125)。结论 N6AMT1在抗氧化中发挥重要作用;在砷代谢中,As3MT、N6AMT1表达保持动态平衡;长时间、高浓度的砷会造成砷代谢障碍。
[Abstract]:Objective to investigate the effect of arsenic on the expression of arsenic methylation metabolic enzymes in human skin keratinocyte Ha Ca T cells silenced by Keap1 gene. The gene expression of nuclear transcription factor erythrocytic line -2p45NF-E2) related to KELCH-like epichlorohydrin-associated protein 1keap1and trivalent arsenic methyltransferase (As3MTMT-N-6adenine DNA methyltransferase 1N6AMT1) was detected by real-time fluorescence quantitative PCR(q. Enzyme linked immunosorbent assay (ELISAA) was used to detect the protein concentration of homocysteine cysteine (HCYC), S-adenosine methionine (SAM), as3MTN6AMT1. Results the results of QPCR showed that: compared with group A, Inhibition of Keap1 gene up-regulated the expression of N6AMT1 gene in cells at 8h and 24h and 48h. Compared with group B, the expression of N6AMT1 gene up-regulated P0.0177h, N6AMT1 gene expression down-regulated P0.0177h, N6AMT1 gene expression down-regulated P0.0177h and N6AMT1 gene expression up-regulated P0.0178hAs3MT48h compared with group B. When arsenic concentration was 29 渭 mol/L, the expression of N6AMT1 gene down-regulated P0.017AMT1 gene and up-regulated the expression of P0.017AMT1 gene at 29 渭 mol/L. The results of Elisa showed that the expression of N6AMT1 gene was higher than that of group B. Arsenic concentration 鈮，

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