Qingxuan Tongluo Formula regulates TRAF6 gene through exosomal miRNA-146a regulating inflammatory injury induced by mycoplasma pneumoniae infection in alveolar epithelial cells
[Objective]To investigate the mechanism of Qingxuan Tongluo Formula in regulating Mycoplasma Pneumoniae(MP)infection in alveolar epithelial cells induced inflammatory injury based on the regulation of Tumor necrosis factor receptor-related factor-6(TRAF6)gene by exosomal miRNA-146a.[Methods]Total 24 experimental mice were randomly divided into blank group,Mycoplasma pneumoniae pneumonia(MPP)group,azithromycin group,and Qingxuan Tongluo Formula group.After the end of administration,the body temperature and cough of the mice were observed,and the HE staining,pathological score and PCR identification of lung tissue were performed.Alveolar epithelial cells are divided into normal control group,MP infection group,miRNA-146a overexpression/inhibitor group,miRNA-NC negative control group.RT-qPCR was used to detect miRNA-146a in exosomes,lung tissues of animal experimental mice,and TRAF6 mRNA expression levels in alveolar epithelial cells.Elisa was used to detect Interleukin-17(IL-17),Interleukin-6(IL-6)and Tumor necrosis factor-α(TNF-α)contents in culture medium supernatant.[Results]Compared with the blank group,tthe MPP group had an increase in body temperature,an increase in the number of coughs,a shortened cough latency,and an increase in lung histopathological scores(all P<0.01).Compared with the MPP group,the body temperature of the mice in the Qingxuan Tongluo Formula group decreased,the number of coughs decreased,the cough latency increased,and the lung histopathological score decreased(all P<0.01).Compared with normal control group,the expression level of miRNA-146a in exosomes decreased in MP infection group,and the contents of inflammatory factors as IL-17,IL-6 and TNF-α in alveolar epithelial cells and TRAF6 mRNA in culture medium supernatant were increased(all P<0.05).Compared with the MP infection group,the level of miRNA-146a in the miRNA-146a overexpression group was significantly up-regulated in exosomes,and the expression levels of TRAF6 mRNA in alveolar epithelial cells and IL-17,IL-6 and TNF-α in culture medium supernatant were decreased(all P<0.05).In the miRNA-146a inhibition group,the expression levels of miRNA-146a in exosomes were significantly decreased,and the expression levels of IL-17,IL-6 and TNF-α in culture medium supernatant and TRAF6 mRNA in culture medium supernatant were significantly increased(all P<0.05).Compared with the miRNA-146a inhibitor group,the level of inflammatory factors in the supernatant of the exosome intervention group in the Qingxuan tongluo Formula group was significantly reduced(all P<0.01).Compared with the MPP group,the content of miRNA-146a in the lung tissue of mice in the Qingxuan Tongluo Formula group was increased(P<0.01).[Conclusion]Qingxuan Tongluo formula may regulate the inflammatory injury induced by MP infection by up-regulating exosomal miRNA-146a and targeting the TRAF6 gene,which in turn affects the expression of inflammatory factors such as IL-17,IL-6 and TNF-α.
国家科技期刊平台
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