miR-450b-3p/TLR8轴在RSV感染肾细胞后对细胞因子IL-6、IL-10、IL-17和IFN-γ分泌的研究  

Effects of miR-450b-3p/TLR8 axis on the secretion of cytokines IL-6,IL-10, IL-17 and IFN-γ after RSV infection of renal cells

作　　者：吴东 张庆红[1] 胡兆雄[1] 许为佳 杨山珊 WU Dong;ZHANG Qinghong;HU Zhaoxiong;XU Weijia;YANG Shanshan(Department of Nephrology,Taihe Hospital,Affiliated Hospitol of Hubei University of Medicine/Shiyan Taihe Hospital,Shiyan 442000,China)

机构地区：[1]湖北医药学院附属太和医院/十堰市太和医院肾病内科,442000

出　　处：《免疫学杂志》2022年第12期1046-1053,共8页Immunological Journal

摘　　要：目的 探讨呼吸道合胞病毒(RSV)感染后细胞因子风暴引起肾损伤的潜在机制。方法 RSV感染肾细胞ACHN后,检测细胞因子风暴关键分子IL-6、IL-10、IL-17和IFN-γ的分泌水平。通过miRNA高通量测序检测RSV感染后不同时间段ACHN细胞中miRNA水平,并通过Pearson相关系数分析miRNA表达水平与IL-6、IL-10、IL-17和IFN-γ分泌水平的相关性。通过miRNA inhibitor筛选高相关的miRNA与细胞因子的关系。此外,鉴定miRNA的关键底物。结果 RSV感染肾细胞ACHN后,细胞因子IL-6、IL-10、IL-17和IFN-γ的分泌水平随着感染时间增加而升高。干扰miR-450b-3p后,IL-6、IL-10、IL-17和IFN-γ的分泌水平上升。敲低TLR8时,RSV感染肾细胞后IL-6、IL-10、IL-17和IFN-γ的分泌水平下降。过表达miR-450b-3p后,TLR8的mRNA水平和蛋白水平均下降;干扰miR-450b-3p后,TLR8的m RNA水平和蛋白水平均上升。此外,miR-450b-3p靶向TLR8 mRNA的3’端非编码区。TLR8敲除后,RSV感染时过表达miR-450b-3p,IL-6、IL-10、IL-17和IFN-γ的分泌水平无显著变化。过表达miR-450b-3p后,MyD88与TRAF6的相互作用水平下降,IRF7和p65的磷酸化水平下降;干扰miR-450b-3p后,MyD88与TRAF6的相互作用水平上升,IRF7和p65的磷酸化水平上升。结论 miR-450b-3p靶向TLR8 mRNA的3’端非编码区,减少了TLR8的蛋白表达水平。RSV感染肾细胞后,miR-450b-3p水平下降,TLR8水平上升,MyD88与TRAF6的相互作用水平上升,转录因子IRF7和p65被激活,细胞因子IL-6、IL-10、IL-17和IFN-γ的分泌水平升高。It has been found that respiratory syncytial virus(RSV) can aggravate primary nephrotic syndrome,in which cytokine-mediated inflammation plays a key role. The purpose of this study was to investigate the potentialmechanism of cytokine storm-induced renal injury after RSV infection. In this study, we found that the secretionlevels of cytokines IL-6, IL-10, IL-17 and IFN-γ increased with the infection time after RSV infected ACHN cells;mir-450b-3p interference also promoted the secretion of IL-6, IL-10, IL-17 and IFN-γ. When TLR8 was knockeddown, the secretion levels of IL-6, IL-10, IL-17 and IFN-γ decreased in RSV-infected renal cells. Afteroverexpression of mir-450b-3p, the mRNA and protein levels of TLR8 decreased. Both mRNA and protein levels ofTLR8 increased after mir-450b-3p interference. In addition, mir-450B-3p targets the 3 ’non-coding region ofTLR8 mRNA. After TLR8 knockout, mir-450b-3p overexpression can not induce any significant change in thesecretion levels of IL-6, IL-10, IL-17 and IFN-γ. After overexpression of mir-450b-3p, the interaction level ofMyD88 with TRAF6 decreased, and the phosphorylation level of IRF7 and p65 decreased, while mir-450b-3p interference could reverse these changes. Therefore, mir-450b-3p targets the 3’non-coding region of TLR8 mRNAand reduces the protein expression level of TLR8. RSV infection in renal cells could down-regulate mir-450b-3p level, while up-regulate the levels of TLR8, IL-6, IL-10,IL-17 and IFN-γ, promote the interaction between MyD88 and TRAF6, and activate transcription factors IRF7and p65.

关 键 词：miR-450b-3p TLR8 RSV ACHN 先天免疫 

分 类 号：R373.1[医药卫生—病原生物学]