登革2型病毒在RAW264.7细胞和THP-1细胞中复制能力及免疫激活能力的比较  

Comparison of the Replication Ability and Immune Activation Ability of Dengue 2 Virus in RAW264.7 Cells and THP-1 Cells

作　　者：牟小会 吴家红 张海龙 匡贵榕 MU Xiaohui;WU Jiahong;ZHANG Hailong(Department of Reproductive Medicine,the People’s Hospital of Anshun City,Guizhou Province 561000)

机构地区：[1]贵州省安顺市人民医院生殖医学科,561000 [2]贵州医科大学基础医学院病原生物学实验室 [3]贵州省安顺市人民医院医学检验科

出　　处：《医学理论与实践》2024年第17期2897-2901,共5页The Journal of Medical Theory and Practice

摘　　要：目的:比较登革2型病毒(DENV2)在RAW264.7细胞和THP-1细胞中的复制能力及DENV2对这两种靶细胞的免疫激活作用。方法:将DENV2病毒液与靶细胞(RAW264.7细胞和THP-1细胞)分别孵育6h、12h、24h,孵育结束后收集细胞,Trizol法提取不同时间的细胞总RNA,检测RNA纯度和浓度后逆转录为cDNA,使用qRT-PCR检测样本中病毒目的基因DENV2 E及细胞因子IL-1β、IL-6、TNF-α、IFN-β、ISG15、CCL2的mRNA相对表达量。结果:DENV2感染RAW264.7细胞和THP-1细胞后,DENV2 E基因的表达量均显著上调(P<0.05),且呈现逐渐增加的趋势。6h、12h时,DENV2 E基因在两个靶细胞中的相对表达量一致;24h时,DENV2 E基因在RAW264.7细胞中的相对表达量显著高于THP-1细胞。与未感染DENV2组相比,DENV2感染RAW264.7细胞和THP-1细胞6h、12h、24h后,IL-1β、IL-6、TNF-α、ISG15、IFN-β、CCL2基因的相对表达量均显著升高(P<0.05),且在RAW264.7细胞中的相对表达量显著高于THP-1细胞。结论:DENV2在RAW264.7细胞中的复制能力强于THP-1细胞的复制能力,且DENV2在RAW264.7细胞中具有更强的免疫激活能力。Objective:To compare the replication ability of DENV2 in RAW264.7 cells and THP-1 cells,as well as the immune activation effect of DENV2 on these two target cells.Methods:Raw264.7 cells and THP-1 cells were treated with DENV2 virus solution for 6 hours,12 hours and 24 hours.After incubation,RNA was isolated by using the Trizol method from collecting cells,cDNA was generated and the mRNA levels of inflammatory cytokines(IL-1β,IL-6,TNF-α),type I interferon(IFN-β),antiviral protein(ISG15)and chemokine(CCL2)were detected by qRT-PCR after stimulation.Results:Compared with the uninfected DENV2 group,the expression level of DENV2 E gene was significantly upregulated(P<0.05)in DENV2 infected RAW264.7 cells and DENV2 infected THP-1 cells,which show a gradually increasing trend.At 6 hours and 12 hours,the relative expression levels of DENV2 E gene in the two target cells were consistent;however,the relative expression level of DENV2 E gene in RAW264.7 cells was significantly higher than that in THP-1 cells at 24 hours.After DENV2 infection of RAW264.7 cells and THP-1 cells for 6 hours,12 hours,and 24 hours,the relative expression levels of IL-1β,IL-6,TNF-α,ISG15,IFN-β,CCL2 genes were significantly increased(P<0.05),and the relative expression levels of those genes in RAW264.7 cells were significantly higher than those in THP-1 cells.Conclusion:The replication ability of DENV2 in RAW264.7 cells is stronger than that in THP-1 cells,and DENV2 has stronger immune activation ability in RAW264.7 cells.

关 键 词：DENV2 RAW264.7细胞 THP-1细胞 复制能力 免疫激活 

分 类 号：R-33[医药卫生]