Background

Tuberculosis (TB) is a very serious public health problem in the world at present. The incidence rate is rising continuously. Once it develops to the middle and late stage, it can cause serious tissue damage and necrosis, directly threatening the life and health of patients. Because of its high incidence, high infectivity and high mortality, clinical research on TB has never stopped. Previous studies have confirmed the effect of macrophages on mycobacterium tuberculosis (MTB) infection, and its regulatory mechanism has not yet been fully clarified.

Objective

To analyze the effects of NEAT1 and miR-377–3p on macrophages infected by MTB and provide new reference for the diagnosis and treatment of TB in the future.

Methods

The pulmonary TB patients admitted to our hospital from July 2017 to July 2019 and the healthy cases at the same time were selected as the research objects for prospective analysis. In addition, THP-1 cells and mycobacterium tuberculosis H37Ra were purchased to construct macrophages infected by H37Ra, and the effects of NEAT1 and miR-377–3p on macrophages and their relationship with inflammatory factors were analyzed.

Results

NEAT1 was highly expressed in pulmonary TB patients and miR-377–3p was poorly expressed (p < 0.05). The concentrations of inflammatory factors in serum of patients with pulmonary TB were significantly higher than those in healthy cases (P < 0.001). After infection with H37Ra, the inflammatory factors in macrophages increased significantly (p < 0.001), while miR-377–3p decreased (p < 0.001). Inhibition of NEAT1 and increase of miR-377–3p could decrease inflammatory factors and proliferation ability of cells, and increase apoptosis rate (p < 0.001). The double luciferase reporter assay showed that the fluorescence activity of NEAT1-WT was inhibited by the transfection of miR-377-3pmimics (P < 0.001).

Conclusion

NEAT1 participates in inflammatory response in macrophages infected by MTB through targeted regulation of miR-377–3p.

背景

结核病（TB）是目前世界范围内非常严重的公共卫生问题。发病率不断上升。一旦发展到中晚期，可引起严重的组织损伤和坏死，直接威胁患者的生命和健康。由于其高发病率、高传染性和高死亡率，结核病的临床研究从未停止。既往研究证实了巨噬细胞对结核分枝杆菌（MTB）感染的影响，其调控机制尚未完全阐明。

客观的

分析NEAT1和miR-377-3p对MTB感染巨噬细胞的影响，为今后结核病的诊治提供新的参考。

方法

选取2017年7月至2019年7月我院收治的肺结核患者及同期健康病例作为研究对象进行前瞻性分析。此外，购买THP-1细胞和结核分枝杆菌H37Ra构建H37Ra感染的巨噬细胞，分析NEAT1和miR-377-3p对巨噬细胞的影响及其与炎症因子的关系。

结果

NEAT1 在肺结核患者中高表达，而 miR-377-3p 低表达（p < 0.05）。肺结核患者血清中炎症因子浓度显着高于健康人群（P < 0.001）。感染 H37Ra 后，巨噬细胞中的炎症因子显着增加（p < 0.001），而 miR-377-3p 减少（p < 0.001）。抑制 NEAT1 和增加 miR-377-3p 可以降低炎症因子和细胞增殖能力，增加细胞凋亡率（p < 0.001）。双荧光素酶报告基因检测显示，转染 miR-377-3pmimics 可抑制 NEAT1-WT 的荧光活性（P < 0.001）。

结论

NEAT1 通过靶向调节 miR-377-3p 参与 MTB 感染的巨噬细胞的炎症反应。