Department of Critical Care Medicine, Dezhou People's Hospital, Dezhou, China.
Eur Rev Med Pharmacol Sci. 2018 Sep;22(17):5620-5626. doi: 10.26355/eurrev_201809_15827.
To investigate the possible role of microRNA-218 in the pathogenesis of sepsis and its underlying mechanism.
MicroRNA-218 expression in peripheral blood mononuclear cells (PBMCs) of 53 sepsis patients and 20 healthy controls was detected by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). MicroRNA-218 expression in Treg cells of sepsis patients and healthy controls was also detected. The binding condition of microRNA-218 to VOPP1 was confirmed by dual-luciferase reporter gene assay and RNA binding protein immunoprecipitation (RIP) assay, respectively. Furthermore, sepsis mouse model was constructed. MicroRNA-218 mimics or inhibitor was injected into mouse tail vein, respectively. The proportion of Treg cells was compared between sepsis mice injected with microRNA-218 mimics and inhibitor. Expressions of microRNA-218 and VOPP1 in Treg cells extracted from sepsis mouse were detected. ELISA (enzyme-linked immunosorbent assay) assay was conducted to detect serum levels of inflammatory factors (TNF-α, IL-6, TGF-β, and IL-10) in sepsis mouse. Finally, protein expressions of key genes in JAK/STAT pathway in sepsis mouse spleen were detected by Western blot.
MicroRNA-218 expression in sepsis patients was remarkably lower than that of healthy controls, which was gradually decreased with the deteriorating symptoms. Specifically, microRNA-218 expression was the lowest in patients who died of sepsis. Downregulated microRNA-218 was seen in Treg cells extracted from advanced sepsis patients. Both dual-luciferase reporter gene assay and RIP assay suggested that microRNA-218 can bind to VOPP1. VOPP1 expression was negatively regulated by microRNA-218. In advanced sepsis mouse, administration of microRNA-218 mimics increased expressions of TNF-α and IL-6, but decreased expressions of IL-10 and TGF-β. Western blot results indicated that microRNA-218 can inhibit the JAK/STAT pathway in sepsis mice.
MicroRNA-218 expression in the PBMCs of sepsis patients was remarkably reduced, which inhibited sepsis development via negatively regulating VOPP1 and suppressing JAK/STAT pathway.
探讨 microRNA-218 在脓毒症发病机制中的可能作用及其潜在机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 53 例脓毒症患者和 20 例健康对照者外周血单个核细胞（PBMC）中 microRNA-218 的表达，检测脓毒症患者和健康对照者 Treg 细胞中 microRNA-218 的表达。分别采用双荧光素酶报告基因检测和 RNA 结合蛋白免疫沉淀（RIP）实验验证 microRNA-218 与 VOPP1 的结合情况。此外，构建脓毒症小鼠模型，分别尾静脉注射 microRNA-218 模拟物和抑制剂，比较注射 microRNA-218 模拟物和抑制剂的脓毒症小鼠 Treg 细胞的比例。检测脓毒症小鼠 Treg 细胞中 microRNA-218 和 VOPP1 的表达。采用酶联免疫吸附试验（ELISA）检测脓毒症小鼠血清中炎症因子（TNF-α、IL-6、TGF-β和 IL-10）水平。最后，采用 Western blot 检测脓毒症小鼠脾组织中 JAK/STAT 通路关键基因的蛋白表达。
脓毒症患者 PBMC 中 microRNA-218 的表达明显低于健康对照组，且随着病情恶化逐渐降低，尤其是死于脓毒症的患者表达最低。晚期脓毒症患者 Treg 细胞中出现下调的 microRNA-218。双荧光素酶报告基因检测和 RIP 实验均提示 microRNA-218 可与 VOPP1 结合，且 VOPP1 的表达受 microRNA-218 的负调控。晚期脓毒症小鼠给予 microRNA-218 模拟物后，TNF-α和 IL-6 的表达增加，而 IL-10 和 TGF-β的表达减少。Western blot 结果表明，microRNA-218 可抑制脓毒症小鼠的 JAK/STAT 通路。
脓毒症患者 PBMC 中 microRNA-218 的表达明显降低，通过负向调控 VOPP1 抑制 JAK/STAT 通路，从而抑制脓毒症的发生发展。
