Department of Orthopaedics, Changhai Hospital, Second Military Medical University, Shanghai, China.
Eur Rev Med Pharmacol Sci. 2021 Jan;25(1):65-70. doi: 10.26355/eurrev_202101_24349.
The purpose of this study was to uncover the role of microRNA-665 (miR-665) in protecting inflammatory response in microglia following spinal cord injury (SCI) and the underlying mechanism.
The serum levels of miR-665 and TREM2 (triggering receptor expressed on myeloid 2) in SCI patients (n=24) and healthy subjects (n=24) were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Then, the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). After lipopolysaccharide (LPS) induction in BV2 cells, the relative levels of miR-665 and TREM2 were detected by qRT-PCR, and relative levels of IL-6 and TNF-α in the culture medium were examined by ELISA. Next, TREM2, the target gene of miR-665, was determined by Dual-Luciferase reporter assay, and the relationship between the expression levels of TREM2 and miR-665 in SCI patients and BV2 cells was analyzed. Finally, the regulatory effects of miR-665 and TREM2 on IL-6 and TNF-α levels in the culture medium of LPS-induced BV2 cells were assessed.
It was found that miR-665 was downregulated in serum of SCI patients and LPS-induced BV2 cells, while TREM2 was upregulated. Silenced miR-665 or overexpressed TREM2 was involved in protecting inflammatory response following SCI. Besides, rescue experiments showed that miR-665 participated in the regulation of inflammatory response following SCI by targeting TREM2.
MiR-665 inhibits inflammatory response following SCI by targeting TREM2.
本研究旨在揭示 microRNA-665（miR-665）在脊髓损伤（SCI）后小胶质细胞炎症反应中的保护作用及其机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 24 例 SCI 患者和 24 例健康对照者血清 miR-665 和 TREM2（髓样细胞触发受体 2）水平，酶联免疫吸附试验（ELISA）检测白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平。用脂多糖（LPS）诱导 BV2 细胞后，qRT-PCR 检测 miR-665 和 TREM2 的相对水平，ELISA 检测培养上清中 IL-6 和 TNF-α的相对水平。双荧光素酶报告基因检测 TREM2 作为 miR-665 的靶基因，分析 SCI 患者和 BV2 细胞中 TREM2 和 miR-665 表达水平的关系。最后，评估 miR-665 和 TREM2 对 LPS 诱导的 BV2 细胞培养上清中 IL-6 和 TNF-α水平的调节作用。
发现 miR-665 在 SCI 患者血清和 LPS 诱导的 BV2 细胞中下调，而 TREM2 上调。沉默 miR-665 或过表达 TREM2 参与 SCI 后炎症反应的保护。此外，挽救实验表明，miR-665 通过靶向 TREM2 参与 SCI 后炎症反应的调节。
miR-665 通过靶向 TREM2 抑制 SCI 后的炎症反应。
