The Mechanism by Which miR-93-5p Regulates the TLR4 Gene to Inhibit the Inflammatory Response of Bronchial Epithelial Cells Infected by Respiratory Syncytial Virus
This study investigated how miR-93-5p regulates the TLR4 gene in respiratory syncytial virus(RSV)infection of bronchial epithelial cells.Human 16-HBE bronchial epithelial cells were cultured and infected with RSV.Cells were divided into groups:RSV+miR-NC,RSV+miR-93-5p,RSV+si-NC,RSV+si-TLR4,RSV+pcDNA-NC,RSV+pcDNA-TLR4,RSV+miR-93-5p+pcDNA-NC,and RSV+miR-93-5p+pcDNA-TLR4.Real-time fluorescent quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to measure the expression levels of miR-93-5p and TLR4 mRNA;enzyme-linked immunosorbent assay was used to measure the levels of tumor necrosis factor-α(TNF-α)and interleukins(IL)-1β and-6;the Cell Counting Kit(CCK-8)method was used to detect cell proliferation;Western blot was used to measure cyclin 1(CyclinD1)and P21 protein expression;dual luciferase reporter assay was used to verify the targeted regulation of TLR4 by miR-93-5p.Compared with the control group,the expression level of miR-93-5p was significantly decreased in the RSV-infected group,and the expression level of TLR4 mRNA was significantly increased(P＜0.05).Compared with the RSV+miR-NC group,the expression level of miR-93-5p in 16-HBE cells in the RSV+miR-93-5p group was significantly increased;the levels of TNF-α,IL-1β,and IL-6 were significantly decreased;the cell survival rate and CyclinD1 protein expression increased significantly;and P21 protein expression decreased significantly(P＜0.05).Compared with the RSV+si-NC group,the expression level of TLR4 mRNA was significantly decreased in the 16-HBE cells of the RSV+si-TLR4 group;the levels of TNF-α,IL-1β,and IL-6 were significantly decreased;the cell survival rate and CyclinD1 protein expression were significantly increased;and the expression of P21 protein decreased significantly(P＜0.05).Compared with the RSV+miR-93-5p+pcDNA-NC group,the expression level of TLR4 mRNA in the 16-HBE cells in the RSV+miR-93-5p+pcDNA-TLR4 group increased significantly;the levels of TNF-α,IL-1β,and IL-6 increased significantly;the cell survival rate and CyclinD1 protein expression decreased significantly;and P21 expression increased significantly(P＜0.05).MiR-93-5p negatively regulates the TLR4 gene to inhibit the inflammatory response of bronchial epithelial cells infected by RSV.
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