六溴环十二烷对视黄醛类X受体α、孕烷X受体、过氧化物酶体增殖物活化受体γ的影响及其相互作用

发布时间：2018-03-31 13:57

  本文选题：六溴环十二烷　切入点：视黄醛类X受体α　出处：《卫生研究》2017年03期

【摘要】：目的探讨六溴环十二烷(HBCDs)对小鼠神经母细胞瘤细胞N2a增殖的影响以及对3个重要细胞核受体:视黄醛类X受体α(RXRα)、过氧化物酶体增殖物活化受体γ(PPARγ)、孕烷X受体(PXR)表达及其相互作用的影响。方法用不同浓度的3种非对映异构体(±)α-HBCD,(±)β-HBCD,(±)γ-HBCD处理N2a细胞,Cell counting kit-8(CCK-8)法检测HBCD对N2a的细胞毒性作用;流式细胞术检测HBCD对N2a细胞周期的影响;实时荧光定量(RT-PCR)和免疫蛋白印迹(Western blot,WB)法分别用于检测3个细胞核受体RXRα、PPARγ、PXR和下游靶基因细胞色素P450亚酶CYP3A11的mRNA及蛋白表达水平的变化;免疫共沉淀技术分析RXRα、PXR、PPARγ受体间的相互作用。结果β-HBCD对N2a的细胞毒性明显大于α-HBCD,γ-HBCD没有明显的细胞毒性。α-HBCD、β-HBCD对N2a细胞增殖的抑制作用呈时间-剂量效应关系(P0.05),其半数抑制浓度(IC_(50))分别为60.07和10.52μmol/L,γ-HBCD的细胞毒性较小,镜下可见黑色絮状物,CCK-8法未能测定出其IC_(50);α-、β-HBCD会使细胞周期阻滞在G2/M期;染毒24 h后,RXRα、PPARγ、PXR及CYP3A11的mRNA及蛋白表达水平均呈现上升趋势(P0.05);在N2a细胞内,α-HBCD染毒前后,RXRα与PPARγ、PXR之间始终存在交互作用关系。结论α-HBCD、β-HBCD对N2a细胞具有增殖抑制作用,细胞周期主要阻滞在G2/M期。α-HBCD、β-HBCD均可诱导3种细胞核受体RXRα、PPARγ和PXR的表达升高,PXR受体下游表达基因CYP3A11的表达也明显升高(P0.05)。RXRα与PPARγ、PXR三个细胞核受体之间始终存在交互作用,但是受体间相互作用的分子机制有待深入研究。
[Abstract]:Objective to investigate the effect of HBCDs on the proliferation of mouse neuroblastoma cell line N2a, and to investigate the effects of HBCDs on the proliferation of neuroblastoma cells N2a and on three important nuclear receptors: Retinal X receptor 伪 (RXR 伪), peroxisome proliferator activated receptor 纬 (PPAR 纬) and pregnancy X receptor (PXR). Methods the cytotoxic effects of HBCD on N2a cells were detected by different concentrations of (卤) 伪 -HBCD- (卤) 尾 -HBCD- (卤) 纬 -HBCD-treated N2a cells. The effect of HBCD on N2a cell cycle was detected by flow cytometry, and the changes of mRNA and protein expression of RXR 伪 -PPAR- 纬 PXR and cytochrome P450 subenzyme CYP3A11 were detected by real-time fluorescence quantitative RT-PCR and Western blotblotWB assay, respectively. Results the cytotoxicity of 尾 -HBCD to N2a was significantly greater than that of 伪 -HBCD, but 纬 -HBCD had no obvious cytotoxicity. The inhibitory effects of 伪 -HBCD and 尾 -HBCD on the proliferation of N2a cells were time-dose dependent (P0.05A). The concentration of IC-HBCD was 60.07 渭 mol / L and 10.52 渭 mol / L, respectively. The cytotoxicity of 纬 -HBCD was less. The results showed that the black flocculant CCK-8 method could not determine its ICS 50, 伪 -, 尾 -HBCD could block the cell cycle in G _ 2 / M phase. After 24 hours of exposure, the expression of mRNA and protein in RXR 伪 PPAR- 纬 PXR and CYP3A11 showed an increasing trend (P0.05A), and there was always interaction between RXR 伪 and PPAR 纬 PXR in N2a cells before and after exposure to 伪 -HBCD. Conclusion 伪 -HBCD, 尾 -HBCD can inhibit the proliferation of N2a cells, and 尾 -HBCD can inhibit the proliferation of N2a cells. The cell cycle was mainly blocked in G _ 2 / M phase. 伪 -HBCD and 尾 -HBCD could induce the expression of RXR 伪 -PPAR 纬 and PXR in three nuclear receptors. The expression of CYP3A11 gene downstream of PXR receptor was also significantly increased. There was always interaction between the three nuclear receptors of PPAR 纬 -PXR and PXR 伪. However, the molecular mechanism of receptor interaction needs further study.
【作者单位】： 广州医科大学公共卫生学院;深圳市疾病预防控制中心;四川大学公共卫生学院;
【基金】：国家自然科学基金(No.21677103) 广东省科技厅产业技术研究与开发资金(No.2013B030800001)
【分类号】：R114
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本文编号：1691038