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目的:探讨2,2’,4,4’-四溴联苯醚(BDE-47)对人神经母细胞瘤SK-N-SH细胞增殖及对RXRα、TRs、PPARs核受体表达水平的影响,为阐明多溴联苯醚化合物神经毒性作用提供科学依据。方法:采用CCK-8法检测不同浓度BDE-47处理不同时间后的SK-N-SH细胞增殖情况;再以5、10和20μmol/L BDE-47染毒细胞24 h后,分别采用DCFH-DA荧光法、WST-1法、比色法、qPCR法测定胞内ROS水平、SOD活力、GSH-Px活力及hOGG1 m RNA表达量;采用qPCR及Western blot检测BDE-47对RXRa、TRs、PPARs受体m RNA及蛋白表达水平的影响。结果:BDE-47抑制SK-N-SH细胞增殖,其抑制作用呈明显的时间和剂量效应关系(P<0.05),24 h半数抑制浓度(IC_(50))为75.94μmol/L;与溶剂对照组相比,10和20μmol/L BDE-47导致细胞内ROS水平上升、SOD活力下降、GSH-Px活力下降、hOGG1 m RNA水平上升,并可致SK-N-SH细胞氧化损伤(P<0.05或P<0.01);BDE-47可诱导RXRα、TRs及PPARs受体各亚型m RNA及蛋白表达显著上升(P<0.05或P<0.01),并且对各亚型的诱导程度不一。结论:BDE-47抑制人神经母细胞瘤SK-N-SH细胞的增殖,导致氧化损伤,上调RXRα、TRs、PPARs核受体的表达从而介导神经毒性作用。
AIM: To investigate the effects of 2,2 ’, 4,4’-tetrabromodiphenyl ether (BDE-47) on the proliferation of human neuroblastoma SK-N-SH cells and the expression of nuclear receptor RXRα, TRs and PPARs , To provide a scientific basis for clarifying the neurotoxic effects of polybrominated diphenyl ether compounds. Methods: The proliferation of SK-N-SH cells treated with different concentrations of BDE-47 was detected by CCK-8 method. After treated with 5, 10 and 20 μmol / L BDE-47 for 24 h, DAF, WST-1 method, colorimetric method and qPCR method were used to determine intracellular ROS level, SOD activity, GSH-Px activity and hOGG1 m RNA expression; qPCR and Western blot were used to detect the expression of RXRa, TRs and PPARs Receptor m RNA and protein expression levels. Results: BDE-47 inhibited the proliferation of SK-N-SH cells in a dose-dependent and time-dependent manner (P <0.05), and the IC50 value of IC50 was 75.94μmol / L Compared with the control group, 10 and 20 μmol / L BDE-47 resulted in an increase in intracellular ROS levels, decreased SOD activity, decreased GSH-Px activity, increased hOGG1 m RNA levels and oxidative damage in SK-N-SH cells (P < 0.05 or P <0.01). BDE-47 induced a significant increase of mRNA and protein expression of RXRα, TRs and PPARs subtypes (P <0.05 or P <0.01). CONCLUSION: BDE-47 inhibits the proliferation of human neuroblastoma SK-N-SH cells, resulting in oxidative damage and upregulation of nuclear receptors of RXRα, TRs and PPARs to mediate neurotoxic effects.