Department of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, 89668Tongji University, China.
Innate Immun. 2022 Jan;28(1):11-18. doi: 10.1177/17534259211064285. Epub 2021 Dec 3.
Macrophage autophagy plays a major role in the control and elimination of invading . However, the function and mechanism of circRNA on macrophage autophagy in tuberculosis remain unclear. Therefore, this study aimed to explore the role of circRNA underlying macrophage autophagy in tuberculosis. Quantitative real-time polymerase chain reaction was used to detect the expression of hsa_circ_0045474, miR-582-5p and TNKS2. Autophagy was detected by LC3B immunofluorescence and transmission electron microscopy. Dual-luciferase reporter assays were used to detect the relationship of miR-582-5p and hsa_circ_0045474 or TNKS2. Western blot was used to detect the expression of LC3-І and LC3-ІІ. The results showed that hsa_circ_0045474 was down-regulated in monocytes from patients with tuberculosis and induced autophagy in macrophages. hsa_circ_0045474 sponged miR-582-5p and negatively regulated miR-582-5p expression. Overexpression of miR-582-5p affected by hsa_circ_0045474 induced autophagy in macrophages. TNKS2 served as a target of miR-582-5p and down-regulation of TNKS2 induced autophagy in macrophages regulated by miR-582-5p. In conclusion, our results demonstrated that hsa_circ_0045474 down-regulation induced macrophage autophagy in tuberculosis via miR-582-5p/ TNKS2 axis, implying a novel strategy to treat the occurrence of active pulmonary tuberculosis caused by immune escape of .
巨噬细胞自噬在控制和清除入侵病原体中起着重要作用。然而,circRNA 在结核病中对巨噬细胞自噬的功能和机制尚不清楚。因此,本研究旨在探讨circRNA 在结核病中调控巨噬细胞自噬的作用。采用实时定量聚合酶链反应检测 hsa_circ_0045474、miR-582-5p 和 TNKS2 的表达。通过 LC3B 免疫荧光和透射电镜检测自噬。双荧光素酶报告实验检测 miR-582-5p 与 hsa_circ_0045474 或 TNKS2 的关系。Western blot 检测 LC3-І 和 LC3-ІІ 的表达。结果表明,结核患者单核细胞中 hsa_circ_0045474 下调,并诱导巨噬细胞自噬。hsa_circ_0045474 海绵吸附 miR-582-5p,并负调控 miR-582-5p 的表达。hsa_circ_0045474 过表达对 miR-582-5p 的影响诱导巨噬细胞自噬。TNKS2 是 miR-582-5p 的靶基因,下调 TNKS2 诱导 miR-582-5p 调控的巨噬细胞自噬。总之,我们的研究结果表明,hsa_circ_0045474 下调通过 miR-582-5p/TNKS2 轴诱导结核病巨噬细胞自噬,为治疗因免疫逃避而导致的活动性肺结核提供了新策略。