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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。
OBJECTIVE: To investigate the effect of environmental endocrine disruptors perfluorooctane suifonate (PFOS) on BV-2 and its inflammatory response in mouse microglial cells. Methods: The microglia cells cultured in vitro were exposed to PFOS at different concentrations and time. The viability of BV-2 cells was detected by MTT assay. The expressions of inducible nitric oxide synthesis, iNOS), interleukin (IL) -6 and tumor necrosis factor (TNF) -αmRNA. IL-6 was detected by ELISA, and the expression of nuclear factor-kappa B (NF-κB) was detected by Western blotting. Results: After exposure to different concentrations of PFOS for 12 or 24 hours, cell viability decreased and BV-2 cells were exposed to a significant cell injury. The results of ELISA showed that after the action of PFOS for 24 hours, the secretion of IL-6 increased. In addition, PFOS can cause iNOS, IL-6 gene expression upregulation, while TNF-αexpression has a downward trend. After PFOS incubated with cells, the inflammatory pathway NF-κB was significantly activated, showing an increase in the degree of phosphorylation. The expression of p-NF-κB increased with the increase of exposure concentration. In addition, the expression of p-NF-κB also increased with the increase of exposure time. CONCLUSION: PFOS can decrease the viability of BV-2 cells, activate the NF-κB pathway and promote the release of inflammatory cytokines. This pathway may provide a theoretical basis for elucidating the molecular mechanism of nervous system injury caused by PFOS.