miR-199a调控NF-κB通路在肺炎支原体免疫发病中的作用研究

Role of miR-199a in regulation of NF-κB pathway in immunopathogenesis of mycoplasma pneumoniae

目的:探索miR-199a调控核因子-κB(NF-κB)通路在肺炎支原体免疫发病中的作用。方法:将BALB/c小鼠分为对照组、模型组、miR-199a agomir组、miR-199a agomir阴性对照组、miR-199a antagomir组、miR-199a antagomir阴性对照组(n=10)。以HE检测各组小鼠肺部病理变化情况,行Holfbauer评分,流式细胞术检测外周血中T淋巴细胞CD4+和CD8+比例;ELISA法检测血清TNF-α、IL-6、IL-10、IFN-γ、IgG、IgM、补体C3含量;以qRT-PCR法检测肺组织miR-199a水平;以Western blot检测肺组织NF-κB通路相关蛋白(p-IKK/IKK、p-NF-κB/NF-κB)表达。结果:与模型组相比,miR-199a agomir组小鼠miR-199a、IL-10及C3水平、CD4+/CD8+均升高(P<0.05),Holfbauer评分、TNF-α、IL-6、IFN-γ、IgG及IgM水平、p-IKK/IKK、p-NF-κB/NF-κB表达均降低(P<0.05);miR-199a antagomir组小鼠各指标变化趋势与miR-199a agomir组相反。结论:miR-199a调节肺炎支原体免疫发病,上调其表达可缓解其病情进展,可能是通过抑制NF-κB通路,减轻免疫炎症反应实现的。

Objective:To explore the role of miR-199a in regulating nuclear factor-κB(NF-κB)pathway in immunopathogenesis of mycoplasma pneumoniae.Methods:BALB/c mice were divided into control group,model group,miR-199a agomir group,miR-199a agomir negative control group,miR-199a antagomir group and miR-199a antagomir negative control group(n=10).Pathological changes in each group was detected by HE staining and Holfbauer score was given,percentage of CD4+and CD8+of T lymphocyte in peripheral blood was detected by flow cytometry;serum levels of TNF-α,IL-6,IL-10,IFN-γ,IgG,IgM and complement C3 were detected by ELISA;level of miR-199a in lung tissue was detected by qRT-PCR;expressios of NF-κB pathway related proteins(p-IKK/IKK,p-NF-κB/NF-κB)in lung tissue were detected by Western blot.Results:Compared with model group,levels of miR-199a,IL-10 and C3,CD4+/CD8+were increased(P<0.05),while Holfbauer score,levels of TNF-α,IL-6,IFN-γ,IgG and IgM,p-IKK/IKK,p-NF-κB/NF-κB were decreased in mice of miR-199a agomir group(P<0.05);change trend of each index of mice in miR-199a antagomir group was contrary to that in miR-199a agomir group.Conclusion:miR-199a can regulate the immune pathogenesis of mycoplasma pneumoniae,up-regulate its expression can alleviate progress of the disease,which probably by inhibiting the NF-κB pathway and alleviating the immune inflammatory response.