本文選題：六溴環(huán)十二烷　切入點：視黃醛類X受體α　出處：《衛(wèi)生研究》2017年03期

【摘要】：目的探討六溴環(huán)十二烷(HBCDs)對小鼠神經(jīng)母細胞瘤細胞N2a增殖的影響以及對3個重要細胞核受體:視黃醛類X受體α(RXRα)、過氧化物酶體增殖物活化受體γ(PPARγ)、孕烷X受體(PXR)表達及其相互作用的影響。方法用不同濃度的3種非對映異構(gòu)體(±)α-HBCD,(±)β-HBCD,(±)γ-HBCD處理N2a細胞,Cell counting kit-8(CCK-8)法檢測HBCD對N2a的細胞毒性作用;流式細胞術(shù)檢測HBCD對N2a細胞周期的影響;實時熒光定量(RT-PCR)和免疫蛋白印跡(Western blot,WB)法分別用于檢測3個細胞核受體RXRα、PPARγ、PXR和下游靶基因細胞色素P450亞酶CYP3A11的mRNA及蛋白表達水平的變化;免疫共沉淀技術(shù)分析RXRα、PXR、PPARγ受體間的相互作用。結(jié)果β-HBCD對N2a的細胞毒性明顯大于α-HBCD,γ-HBCD沒有明顯的細胞毒性。α-HBCD、β-HBCD對N2a細胞增殖的抑制作用呈時間-劑量效應(yīng)關(guān)系(P0.05),其半數(shù)抑制濃度(IC_(50))分別為60.07和10.52μmol/L,γ-HBCD的細胞毒性較小,鏡下可見黑色絮狀物,CCK-8法未能測定出其IC_(50);α-、β-HBCD會使細胞周期阻滯在G2/M期;染毒24 h后,RXRα、PPARγ、PXR及CYP3A11的mRNA及蛋白表達水平均呈現(xiàn)上升趨勢(P0.05);在N2a細胞內(nèi),α-HBCD染毒前后,RXRα與PPARγ、PXR之間始終存在交互作用關(guān)系。結(jié)論α-HBCD、β-HBCD對N2a細胞具有增殖抑制作用,細胞周期主要阻滯在G2/M期。α-HBCD、β-HBCD均可誘導(dǎo)3種細胞核受體RXRα、PPARγ和PXR的表達升高,PXR受體下游表達基因CYP3A11的表達也明顯升高(P0.05)。RXRα與PPARγ、PXR三個細胞核受體之間始終存在交互作用,但是受體間相互作用的分子機制有待深入研究。
[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 偽 (RXR 偽), peroxisome proliferator activated receptor 緯 (PPAR 緯) and pregnancy X receptor (PXR). Methods the cytotoxic effects of HBCD on N2a cells were detected by different concentrations of (鹵) 偽 -HBCD- (鹵) 尾 -HBCD- (鹵) 緯 -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 偽 -PPAR- 緯 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 偽 -HBCD, but 緯 -HBCD had no obvious cytotoxicity. The inhibitory effects of 偽 -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 緯 -HBCD was less. The results showed that the black flocculant CCK-8 method could not determine its ICS 50, 偽 -, 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 偽 PPAR- 緯 PXR and CYP3A11 showed an increasing trend (P0.05A), and there was always interaction between RXR 偽 and PPAR 緯 PXR in N2a cells before and after exposure to 偽 -HBCD. Conclusion 偽 -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. 偽 -HBCD and 尾 -HBCD could induce the expression of RXR 偽 -PPAR 緯 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 緯 -PXR and PXR 偽. However, the molecular mechanism of receptor interaction needs further study.
【作者單位】： 廣州醫(yī)科大學(xué)公共衛(wèi)生學(xué)院;深圳市疾病預(yù)防控制中心;四川大學(xué)公共衛(wèi)生學(xué)院;
【基金】：國家自然科學(xué)基金(No.21677103) 廣東省科技廳產(chǎn)業(yè)技術(shù)研究與開發(fā)資金(No.2013B030800001)
【分類號】：R114
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本文編號：1691038