【文献解读】LPS在CHME-5小胶质内介导的TLR4神经炎症信号
2017/12/18 创新医学网

    

    

     文章来源:欧尔意 OAE 微信号

     作者:Sofina

    

     原文出处

     Figueroa-Hall LK, Anderson MB, Das S, Stevens CW, Davis RL. LPS-induced TLR4 neuroinflammatory signaling inCHME-5 microglial cells. Neuroimmunol Neuroinflammation 2017;4:219-31.

     DOI:10.20517/2347-8659.2017.38

     摘要

     目的:在神经炎症领域,由于获得和培养原代的小胶质细胞相对困难且花费较多,确定药物和其他因素的特定效用往往很棘手。因此永生化的小胶质细胞系具有重要价值,但只有有限数量的细胞被确定具有炎症反应信号的特性。因此,脂多糖(lipopolysaccharide,LPS)在CHME-5小胶质细胞系内诱导toll样受体4(toll-like receptor 4,TLR4)信号这一特性有望在中枢神经系统(central nervous system, CNS)中作为炎症信号实验模型来发挥重要价值。方法:最近有人提出CHME-5细胞系来源于小鼠而非人类,因此,本研究应用串联重复基因序列技术,连同免疫印迹、逆转录聚合酶链反应技术以及免疫化学技术来证实CHME-5保留着原代小胶质细胞的形态、表型和功能特性以确证这一说法。结果:LPS可以诱导抑制因子κB-α和核因子Kappa B(NF-κB)p65活化、NF-κB p65的结合活性以及肿瘤坏死因子α基因的表达。此外,研究结果同样证实了小胶质细胞表型保持,正如看到的68基因分化和蛋白表达、免疫荧光增强,同时缺乏GFAP免疫反应一样。TLR4基因表达和免疫荧光在LPS处理后显著升高。结论:以上数据表明CHME-5不是人源的,但确是研究小胶质细胞炎症反应信号的有利工具。

     Aim: In the field of neuroinflammation, identifying specific effects of pharmacological agents and other factors is problematic given the relative difficulty and expense in obtaining and culturing primary microglia. Immortalized microglial cell lines are very useful, but only a limited number have been characterized for inflammatory signaling. Therefore, characterization of lipopolysaccharide (LPS)-induced toll-like receptor 4 (TLR4) signaling in CHME-5, a microglial cell line, is expected to be of value as an experimental model of inflammatory signaling in the central nervous system (CNS). Methods: It was recently suggested that CHME-5 cells are of rat origin, not human, hence, verification of this claim using short tandem repeat genotype sequencing, along with immunoblotting, reverse transcription-polymerase chain reaction, and immunocytochemistry techniques to validate that CHME-5 retain morphological, phenotypical, and functional characteristics of primary microglia were undertaken. Results: LPS induced inhibitor kappa B-alpha and nuclear factor-kappa B (NF-κB) p65 activation, NF-κB p65 binding activity, and tumor necrosis factor alpha gene expression. Additionally, results also confirmed the maintenance of microglial phenotype as seen with increased cluster of differentiation 68 gene and protein expression, immunofluorescence, and the absence of glial fibrillary acidic protein-immunoreactivity. TLR4 gene expression and immunofluorescence were significantly increased after LPS treatment. Conclusion: These data demonstrate that CHME-5 cells are not human, but are indeed a beneficial tool for studying microglial inflammatory signaling.

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     *中文摘要由由编委,来自首都医科大学附属北京同仁医院神经内科,王佳伟教授团队李琳负责翻译。中文内容仅供参考,一切以英文原版为准。

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