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探讨人偏肺病毒感染BALB/c小鼠后肺组织Toll样受体(TLRs)表达变化及PolyI:C对病毒复制的影响。实验小鼠分为hMPV感染组、polyI:C+hMPV组、polyI:C+DMEM组、DMEM对照组,均在滴鼻后1、3、5、7、9和16d处死,取肺组织用于病毒滴度测定、病理检查、通过RT-PCR和实时荧光定量PCR方法检测小鼠肺组织内TLRs mRNA的表达。结果显示:①与hMPV感染组相比,polyI:C+hMPV组小鼠肺内病毒复制水平明显减低,仅在感染后第5d及第7d检测到病毒;病理检查结果亦提示肺部炎症明显减轻。②RT-PCR结果提示:hMPV感染组小鼠与DMEM对照组相比,肺组织内大部分TLRs mRNA表达均升高。③实时荧光定量PCR结果提示:hMPV感染后小鼠肺组织TLR7-8 mRNA增高最为显著,且有时间依赖性;滴鼻后24h,polyI:C+hMPV组TLR3的表达明显升高。提示:人偏肺病毒感染BALB/c小鼠后,可上调小鼠肺组织Toll样受体表达,其中TLR7/8途径可能在启动天然免疫应答中起着重要作用;polyI:C可通过早期活化TLR3,抑制hMPV在小鼠肺内的复制,并减轻肺部炎症。
To investigate the expression of Toll-like receptors (TLRs) in lung tissue of BALB / c mice infected with human metapneumovirus and the effect of PolyI: C on virus replication. The experimental mice were divided into hMPV infection group, polyI: C + hMPV group, polyI: C + DMEM group and DMEM control group. All mice were sacrificed at 1,3,5,7,9 and 16 days after nasal infusion. Virus titer and pathological examination. The expression of TLRs mRNA in lung tissue of mice was detected by RT-PCR and real-time fluorescence quantitative PCR. The results showed that: (1) Compared with hMPV-infected group, the virus in the lungs of polyI: C + hMPV group was significantly lower than that of hMPV-infected group, and the virus was detected only on the 5th and 7th day after infection. Pathological examination also showed that lung inflammation was significantly reduced . ② The results of RT-PCR indicated that the expression of most TLRs mRNA in lung tissue of hMPV-infected mice was higher than that of DMEM control group. Real-time quantitative PCR results showed that the expression of TLR7-8 mRNA in lung tissue of hMPV-infected mice was the most significant and time-dependent. The expression of TLR3 in poly I: C + hMPV group was significantly increased after hMPV infection. Tip: Human metapneumovirus infection of BALB / c mice can up-regulate Toll-like receptor expression in mouse lung tissue, of which TLR7 / 8 pathway may play an important role in initiating innate immune responses; poly I: C can be activated early TLR3, inhibits hMPV replication in mouse lungs and reduces lung inflammation.