Abstract
Expression of Toll-like receptor (TLR)4 and its downstream substances, myeloid differentiation factor 88 (MyD88), NF-κB p65, tumor necrosis factor-α (TNF-α) and GR in human alveolar epithelial cells type Ⅱ (AEC Ⅱ) infected with respiratory syncytial virus (RSV) were investigated, and the antiviral immune mechanism mediated by TLR4 was explored. Human AEC Ⅱ were divided into TLR4-/- group, normal group and TLR4+ group, and also into control group, RSV group and RSV+MP (methylprednisolone) group. MTT assay was used to measure the survival of cells after TLR4 knockout and overexpression, and the survival of normal cells after treatment with MP. The concentration of TLR4, MyD88, NF-κB p65, TNF-α, and GR was measured by ELISA after TLR4 knockout and overexpression. Reverse transcription-quantitative PCR (RT-qPCR) was used to measure the mRNA expression of the gene knockout and overexpression groups. RT-qPCR and western blot analysis were used to determine the expression of TLR4, MyD88, NF-κB p65 and GR in RSV and RSV+MP groups. The concentration of the detected substances in the TLR4-/- group was significantly lower than that in the normal group (P<0.01 and <0.001), and in the TLR4+ group was significantly higher than that in the normal group (P<0.05, <0.01 and <0.001); the expression of RSV in the TLR4-/- group was significantly higher than that in the normal group (P<0.001), and in the TLR4+ group was significantly lower than that in the normal group (P<0.05). The expression levels of TLR4, MyD88 and NF-κB p65 in the RSV and RSV+MP groups were significantly higher than those in the control group (P<0.05, <0.01 and <0.001), and the increase presented in the RSV+MP group was significantly lower than that in the RSV group (P<0.05 and <0.01). TLR4-mediated antiviral immunity of human AEC Ⅱ can reduce the levels of TLR4, MyD88, NF-κB p65 and TNF-α and increase the level of GR, participating in the immune defense and reducing the damage of the viral epithelial cells of human type Ⅱ alveoli, thus improving human immunity.