本文關(guān)鍵詞： Nrf Keap 砷代謝酶　出處：《現(xiàn)代預(yù)防醫(yī)學(xué)》2017年15期 　論文類型：期刊論文

【摘要】：目的砷對Keap1基因沉默的人皮膚角質(zhì)(Ha Ca T)細(xì)胞砷甲基化代謝酶表達(dá)的影響。方法實(shí)驗(yàn)共五組,A組為正常Ha Ca T細(xì)胞,B、C、D、E組為模型細(xì)胞。A、B組不給砷,C、D、E組給予不同濃度的砷。五組細(xì)胞在8h、24h、48h、72h,用實(shí)時(shí)熒光定量PCR(q PCR)檢測核轉(zhuǎn)錄因子紅胞系-2p45(NF-E2)相關(guān)因子-2(Nrf2)、KELCH樣環(huán)氧氯丙烷相關(guān)蛋白1(Keap1)、三價(jià)砷甲基轉(zhuǎn)移酶(As3MT)、N-6-腺嘌呤DNA甲基轉(zhuǎn)移酶1(N6AMT1)的基因表達(dá);酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA)檢測細(xì)胞內(nèi)同型半胱氨酸(HCY)、S-腺苷甲硫氨酸(SAM)、As3MT、N6AMT1的蛋白濃度。結(jié)果(1)QPCR結(jié)果發(fā)現(xiàn):與A組比,抑制Keap1基因使細(xì)胞中N6AMT1基因表達(dá)在8h、24h、48h表達(dá)上調(diào)(P0.05)。與B組比:砷濃度2.9μmol/L,8h、72h N6AMT1、24h As3MT基因表達(dá)上調(diào)(P0.017),8h、72h As3MT、24h N6AMT1基因表達(dá)下調(diào)(P0.017);砷濃度5.8μmol/L,8h N6AMT1、48h As3MT基因表達(dá)上調(diào)(P0.017),8h As3MT、48h N6AMT1基因表達(dá)下調(diào)(P0.017)。砷濃度29μmol/L時(shí),24h N6AMT1基因表達(dá)下調(diào)(P0.017),As3MT基因表達(dá)上調(diào)(P0.017)。(2)ELISA結(jié)果發(fā)現(xiàn):與B組比,砷濃度≤5.8μmol/L SAM、HCY蛋白表達(dá)上調(diào)(P0.0125);砷濃度29μmol/L抑制SAM、HCY蛋白的表達(dá)下調(diào)(P0.0125)。結(jié)論 N6AMT1在抗氧化中發(fā)揮重要作用;在砷代謝中,As3MT、N6AMT1表達(dá)保持動(dòng)態(tài)平衡;長時(shí)間、高濃度的砷會(huì)造成砷代謝障礙。
[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，

本文編號：1523919