过表达miR-144-3p下调感染巨噬细胞IL-6 mRNA抑制BCG的存活
Overexpression of miR-144-3p downregulates IL-6 mRNA in infected macrophages to inhibit BCG survival
江升升 1李杨 1李淑萍 2李雪芳 1张叶 1周怀珩 1刘晓倩 1余晓丽1
作者信息
- 1. 武汉轻工大学生命科学与技术学院,武汉 430023
- 2. 亳州学院生物与食品工程学院,亳州 236800
- 折叠
摘要
探究miR-144-3p对巨噬细胞IL-6 mRNA表达量和巨噬细胞免疫能力的影响,为结核诊疗提供理论参考.方法(1)建立BCG感染THP-1巨噬细胞模型,qPCR检测感染不同时间miR-144-3p和IL-6 mRNA表达变化;(2)瞬转miR-144-3p模拟物/抑制剂来过表达/抑制miR-144-3p后,qPCR检测感染不同时间miR-144-3p和IL-6 mRNA表达变化;皮尔森相关系数分析miR-144-3p和IL-6 mRNA的相关性;(3)CCK-8检测瞬转miR-144-3p模拟物/抑制剂后,CCK-8检测BCG感染的巨噬细胞增殖能力;CFU检测BCG存活状态.结果显示:(1)巨噬细胞感染BCG后,miR-144-3p和IL-6 mRNA表达均显著上升(P＜0.05),miR-144-3p表达量在12 h达到峰值,IL-6 mRNA表达量在24 h达到峰值;(2)巨噬细胞感染BCG后,过表达 miR-144-3p 使 IL-6 mRNA 表达显著下降(P＜0.01);抑制 miR-144-3p 使 IL-6 mRNA表达显著上升(P＜0.01);皮尔森相关系数分析结果显示两者存在负相关性;(3)过表达/抑制miR-144-3p不影响巨噬细胞增殖能力,但过表达miR-144-3p能抑制巨噬细胞中BCG的存活.结论:BCG感染巨噬细胞后miR-144-3p和IL-6 mRNA表达均显著上升;过表达/抑制巨噬细胞 miR-144-3p,miR-144-3p可能负调控IL-6 mRNA 表达;过表达 miR-144-3p能抑制BCG存活,进而增强巨噬细胞的先天免疫能力.
Abstract
To investigate the effect of miR-144-3p on macr-ophage IL-6 mRNA expression and macrophage immunocompetence,and to provide theore-tical reference for tuberculosis diagnosis and treatment.The method is establish a BCG-infected THP-1 macrophage model,qPCR to detect changes in miR-144-3p and IL-6 mRNA expression at different times of infection;transiently transfect miR-144-3p mimics/inhibitors to overpress/inhibiel miR-144-3p after the qPCR to detect miR-144-3p and IL-6 mRNA ex-pression changes at different times of infution;Pearson's correlation coefficient analysis the miR-144-3p and IL-6 mRNA corre-lation;CCK-8 to detect the proliferative capacity of BCG-infected macrophages after t-ransiently transfect-ing miR-144-3p was used mimics/inhibitors;and CFU was used to detect BCG survival stat-us.Both miR-144-3p and IL-6 mRNA expression increased significantly(P＜0.05)after ma-crophages were infected with BCG,with miR-144-3p expression peaking at 12 h,and IL-6 mRNA expression peaking at 24 h.Overexpres-sion of miR-144-3p after macrophages were infected with BCG resulted in a significant decrease(P＜0.01)in IL-6 mRNA expression;inhibition of miR-144-3p significantly increased IL-6 m-RNA expression(P＜0.01);there was a negative correlation between the two as analyzed by Pearson's correlation coefficient;overexpression/inhibition of miR-144-3p did not affect the proliferative capacity of macrophages,but over-expression of miR-144-3p inhibited the survival of BCG in macrophages.Both miR-144-3p and IL-6 mRNA expression were significantly increased in BCG-infected macrophages;overexpression/inhibition of miR-144-3p i-n macrophages and miR-144-3p may negatively regulate IL-6 mRNA expression;overexp-ression of miR-144-3p inhibits BCG survival and thus enhances the innate immunity of m-acrophages.
关键词
结核病/BCG/THP-1/巨噬细胞/miR-144-3p/IL-6/mRNAKey words
tuberculosis/BCG/THP-1 macrophages/miR-144-3p/IL-6 mRNA引用本文复制引用
基金项目
武汉市科技局2020年度应用基础前沿科研项目(2020020601012231)
出版年
2024
NETL
NSTL
万方数据