RSV感染小胶质细胞活化Toll样受体3、7介导炎性反应的初步研究

发布时间：2018-01-01 06:12

本文关键词：RSV感染小胶质细胞活化Toll样受体3、7介导炎性反应的初步研究　出处：《安徽医科大学学报》2013年08期 　论文类型：期刊论文

【摘要】：目的探讨呼吸道合胞病毒(RSV)感染小鼠小胶质细胞(BV2)后,Toll样受体3(TLR3)和Toll样受体7(TLR7)的活化及其介导的炎性反应。方法①RSV体外感染BV2,利用免疫荧光技术分别在12、24和36 h检测细胞内RSV-F蛋白。②分别于感染4、8、12、16、24和36 h收集细胞,以未感染细胞作为正常组。用TRIzol提取细胞总RNA,通过半定量RT-PCR法检测TLR3、TLR7的mRNA转录水平的变化。③Western blot法分别检测感染4、8、12、16、24和36 h后的细胞内核转录因子(NF-κB)蛋白水平的变化。④ELISA法检测细胞上清液中肿瘤坏死因子α(TNF-α)含量的变化。结果①荧光显微镜下可以见到FITC标记RSV-F蛋白的绿色荧光随感染时间的延长而增强。②TLR3、TLR7在感染RSV后mRNA水平在观察的时间内表达均升高并呈时间依赖性关系。③各感染时间点NF-κB蛋白水平表达均升高并呈时间依赖性关系。④感染RSV后,细胞上清液中TNF-α含量均升高并呈时间依赖性关系。结论 RSV感染BV2后可以上调TLR3、TLR7,从而使下游细胞炎性因子TNF-α分泌升高,过度的炎性作用可能与神经系统症状的发生相关。
[Abstract]:Objective to investigate the effect of respiratory syncytial virus (RSVV) infection on mouse microglia (BV2). Toll like receptor 3 (TLR3) and Toll like receptor 7 (TLR7) were activated and their inflammatory responses were mediated. The RSV-F protein was detected at 24 h and 36 h after infection. The cells were collected at 24 h and 36 h after infection, respectively. Uninfected cells were used as normal group. Total RNAs were extracted by TRIzol and TLR3 was detected by semi-quantitative RT-PCR method. The changes of mRNA transcription level in TLR7. 3 Western blot assay were used to detect the infection of 4p812m16. Changes of NF-魏 B protein levels in the cell nucleus after 24 and 36 hours. Detection of TNF-伪-TNF- 伪 in the supernatant by Elisa. Results 1 the green fluorescence of FITC labeled RSV-F protein was enhanced with the increase of infection time under fluorescence microscope. The expression of TLR7 increased in a time dependent manner after RSV infection. 3 the expression of NF- 魏 B protein increased in a time dependent manner at all time points. 4After RSV infection. The content of TNF- 伪 in the supernatant was increased in a time dependent manner. Conclusion RSV infection with BV2 can up-regulate TLR3 and TLR7. Thus, the secretion of TNF- 伪 was increased, and the excessive inflammatory effect may be related to the occurrence of nervous system symptoms.
【作者单位】：安徽医科大学微生物学教研室;安徽中医学院中西医结合临床学院;
【基金】：安徽高校省级自然科学研究重点项目资助(编号:KJ2012A152)
【分类号】：R373
【正文快照】： 呼吸道合胞病毒(respiratory syncytial virus,2013-04-12接收RSV)属副黏病毒科,肺炎病毒属,为有包膜非节段的单股负链RNA病毒,是导致世界范围内婴幼儿下呼吸道感染最常见的病原体之一,也是感染老人和免疫低下人群的病原体之一[1]。该病毒侵袭患儿的呼吸道后,主要通过诱导呼

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