摘要
目的探讨miRNA-492、IL-6在肺炎支原体(mycoplasma pneumoniae,MP)免疫发病机制中的作用。方法 (1)以肺炎支原体膜脂蛋白(LAMPS)刺激单核巨噬细胞,用实时-定量PCR法在6h、12h、24h检测单核巨噬细胞系统中的miR-492水平;(2)以不同菌落计数单位的支原体感染单核巨噬细胞,用ELISA法在6h、12h、24h检测上清IL-6的浓度;(3)构建过表达miR-492的慢病毒载体,稳定共转染THP-1细胞,予PMA刺激诱导分化为单核巨噬细胞,用ELISA法检测其IL-6的表达。结果 (1)LAMPS刺激组在6h、12h时间点单核巨噬细胞miR-492表达水平均高于空白对照组,两者之间差异具有统计学意义(P <0.05),24h时间点差异无统计学意义(P>0.05)。(2)不同菌落计数的支原体感染组在12h、24h时间点单核巨噬细胞IL-6表达均高于空白对照组,差异具有统计学意义(P <0.05),6h时间点差异无统计学意义。(3)过表达miR-492慢病毒转染组的单核巨噬细胞IL-6分泌水平高于空载体对照组,差异有统计学意义(P <0.001)。结论 MiR-492可能通过调控单核巨噬细胞系统免疫炎症因子如IL-6的分泌在肺炎支原体感染和发病中起一定作用。
Objective To investigate the potential roles of miRNA-492 and interleukin-6(IL-6)in the immune pathogenic mechanism of Mycoplasma pneumoniae. Methods ①Real-time quantitative PCR was used to determine the miRNA-492 expression in mononuclear phagocyte system at different time points(6,12 and 24 h)after being stimulated with Mycoplasma pneumonia(MP)lipid associated membrane proteins(LAMPs).②The IL-6 expression in the supernate was detected by ELISA at different time points(6,12 and 24 h)after being infected with different colony-counting of MP.③THP-1 cells were co-transfected by high-expression of miR-492 lentiviral vector,and the cells were induced to differentiate into mononuclear phagocytes,then the expressions of IL-6 of mononuclear phagocyte system were detected by ELISA. Results ①The expression of miR-492 in mononuclear phagocytes after being stimulated by LAMPs was higher than that of the blank control group at the time pints of 6 h and 12 h,and there was significant difference between the blank control group and the LAMPs-stimulated group(P<0.05),but there was no statistical difference at 24 h time point(P>0.05).②The expressions of IL-6 of mononuclear phagocytes after being infected with different colony-counting of MP were higher than those of the control group at 12 h and 24 h,and there were statistically significant difference among different colony-counting of MP infected groups and blank control group(P<0.05),but there was no statistical difference at the time point of 6 hour(P>0.05).③The expression of IL-6 in the mononuclear phagocytes after being co-transfected by high-expression of miR-492 lentiviral vector was increased compared to the blank control group,there was statistically significant difference(P<0.001). Conclusion MiR-492 plays a role in the mycoplasma pneumonia infection and disease attack by regulating the immune inflammatory factors such as the secretion of IL-6 in mononuclear phagocyte system.
作者
邓建萍
周卫芳
孙旦
李玉琴
丁莹
骆亚丽
Deng Jianping;Zhou Weifang;Sun Dan;Li Yuqin;Ding Ying;Luo Yali(Department of Infection,Children’s Hospital of Soochow University,Suzhou 215000,Jiangsu,China)
出处
《右江民族医学院学报》
2018年第5期429-432,共4页
Journal of Youjiang Medical University for Nationalities
基金
苏州市科技计划项目(SS201535)
