Department of Intensive Care Unit, Tianjin Medical University General Hospital, Tianjin 300052, China.
Comput Intell Neurosci. 2022 Jun 2;2022:1714041. doi: 10.1155/2022/1714041. eCollection 2022.
Present study is aimed to explore the role of miR-186-5p in sepsis-induced coagulation disorders and molecular mechanisms.
Thirty-four sepsis patients and 34 respiratory infection/pneumonia patients were selected in the present study. Polymicrobial sepsis model was created by cecal ligation and puncture (CLP). The mRNA expression was detected by qRT-PCR. Western blot was utilized to measure protein expression. Thromborel S Reagent was applied to measure the prothrombin time (PT). Platelet count of blood was measured via LH 780. ELISA kits were utilized to evaluate the fibrinogen and PAI-1 concentration.
MiR-186-5p expression was lower and nicotinamide phosphoribosyltransferase (NAMPT) mRNA expression was higher in sepsis patients in contrast to control group. Coagulation time was markedly prolonged and platelet count was markedly decreased in CLP mice. In addition, fibrinogen concentration was obviously lower and PAI-1 concentration was obviously higher in CLP mice. MiR-186-5p mimic obviously decreased coagulation time and PAI-1 concentration, while raised platelet count and fibrinogen concentration. Targetscan predicted miR-186-5p might directly regulates NAMPT, and luciferase reporter assay verified this prediction. In addition, miR-186-5p mimic obviously inhibited the mRNA expression of NAMPT. Knockdown of NAMPT improved coagulation dysfunction in sepsis. Overexpression of NAMPT reversed the improvement effect of miR-186-5p on coagulation dysfunction. MiR-186-5p mimic markedly inhibited NF-B pathway.
MiR-186-5p inhibited sepsis-induced coagulation disorders via targeting NAMPT and inactivating NF-B pathway.
本研究旨在探讨 miR-186-5p 在脓毒症诱导的凝血紊乱中的作用及分子机制。
本研究纳入 34 例脓毒症患者和 34 例呼吸道感染/肺炎患者。采用盲肠结扎穿孔（CLP）建立多微生物脓毒症模型。通过 qRT-PCR 检测 mRNA 表达，Western blot 检测蛋白表达，血栓弹力 S 试剂检测凝血酶原时间（PT），LH 780 检测血小板计数，ELISA 试剂盒检测纤维蛋白原和 PAI-1 浓度。
与对照组相比，脓毒症患者的 miR-186-5p 表达降低，烟酰胺磷酸核糖转移酶（NAMPT）mRNA 表达升高。CLP 小鼠的凝血时间明显延长，血小板计数明显减少。此外，CLP 小鼠的纤维蛋白原浓度明显降低，PAI-1 浓度明显升高。miR-186-5p 模拟物明显缩短凝血时间，降低 PAI-1 浓度，增加血小板计数和纤维蛋白原浓度。Targetscan 预测 miR-186-5p 可能直接调控 NAMPT，荧光素酶报告基因实验验证了这一预测。此外，miR-186-5p 模拟物明显抑制 NAMPT 的 mRNA 表达。在脓毒症中敲低 NAMPT 可改善凝血功能障碍。过表达 NAMPT 逆转了 miR-186-5p 对凝血功能障碍的改善作用。miR-186-5p 模拟物明显抑制 NF-B 通路。
miR-186-5p 通过靶向 NAMPT 抑制 NF-B 通路，抑制脓毒症引起的凝血紊乱。
