PFOS对BV-2细胞的炎性损伤及机制研究

发布时间：2019-07-30 10:03

【摘要】：目的:探讨环境内分泌干扰物全氟辛磺酸(perfluorooctane suifonate,PFOS)对小鼠小胶质瘤细胞BV-2的损伤作用及其炎性反应。方法:利用体外培养的小胶质细胞,给予不同浓度和作用时间的PFOS进行染毒,通过MTT法检测BV-2细胞活力;实时定量PCR的方法观察诱导型一氧化氮合酶(inducible nitric oxide synthesis,i NOS)、白介素(interleukin,IL)-6和肿瘤坏死因子(tumor necrosis factor,TNF)-αm RNA的表达。ELISA法检测炎性因子IL-6,免疫蛋白印迹法检测核转录因子kappa B(nuclear factor-kappa B,NF-κB)等信号蛋白表达情况。结果:不同浓度PFOS对BV-2细胞染毒12、24 h后,细胞活力下降,引起明显的细胞损伤。ELISA结果显示,PFOS作用BV-2 24 h后,细胞炎性因子IL-6分泌增加。此外,PFOS还可引起i NOS、IL-6基因表达上调,而TNF-α表达有下降趋势。PFOS与细胞孵育后,炎性通路NF-κB明显激活,表现为磷酸化程度升高。且随染毒浓度的增加,p-NF-κB的表达有上升趋势;此外,随染毒时间的增加,p-NF-κB的表达亦有上升趋势。结论:PFOS可引起BV-2细胞活力降低,激活NF-κB通路,促进炎性因子的释放,该途径可为阐明PFOS引起的神经系统损伤的分子机制提供理论依据。
[Abstract]:Aim: to investigate the damage and inflammatory response of environmental endocrine disruptor perfluorooctanesulfonic acid (perfluorooctane suifonate,PFOS) to mouse microglioma cell line BV-2. Methods: microglia cultured in vitro were exposed to PFOS at different concentrations and time, and the activity of BV-2 cells was detected by MTT assay. The expression of (inducible nitric oxide synthesis,i NOS), IL-6 and (tumor necrosis factor,TNF-伪 m RNA was observed by real-time quantitative PCR, and the expression of kappa B (nuclear factor-kappa B, NF-魏 B and other signal proteins were detected by enzyme-linked immunosorbent assay (Elisa). Results: BV-2 cells were exposed to different concentrations of PFOS for 12 hours, and the cell activity decreased after 24 hours, resulting in obvious cell injury. Elisa results showed that the secretion of inflammatory factor IL-6 increased after PFOS treatment with BV-2 for 24 hours. In addition, PFOS could up-regulate the expression of I NOS,IL-6 gene, but the expression of TNF- 伪 decreased. After incubation with cells, the inflammatory pathway NF- kappa B was significantly activated, showing an increase in the degree of phosphorylation. With the increase of exposure concentration, the expression of p 鈮，

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