热打击单核细胞来源外泌体中miR-155对肝细胞炎症的影响及其机制  

The influence of exosomal miR-155 derived from monocytes stimulated by heat stress on the inflammatory response of hepatocytes

作　　者：张明[1] 李悦 杨经文[1] 施学智 肖吉彪 童华生 Zhang Ming;Li Yue;Yang Jing-Wen;Shi Xue-Zhi;Xiao Ji-Biao;Tong Hua-Sheng(Department of Critical Care Medicine,the Sixth Affiliated Hospital of Guangzhou Medical University/Qingyuan People's Hospital,Qingyuan,Guangdong 511500,China;Department of Intensive Care Unit,General Hospital of Southern Theatre Command of PLA/Key Laboratory of Trauma and Tissue Repair of Tropical Area of PLA,Guangzhou,Guangdong 510010,China)

机构地区：[1]广州医科大学附属第六医院/清远市人民医院重症医学科一区,广东清远511500 [2]解放军南部战区总医院重症医学科/全军热区损伤与组织修复重点实验室,广东广州510010

出　　处：《解放军医学杂志》2023年第9期1017-1022,共6页Medical Journal of Chinese People's Liberation Army

基　　金：广东省医学科研基金(A2021404);广东省中医药局科研项目(20212292);广东省清远市科技计划项目(200715164560747)。

摘　　要：目的研究热打击单核细胞来源外泌体对肝细胞炎症损伤的影响及其与外泌体miR-155的关系。方法将单核细胞THP-1分为对照组、热打击组、乌司他丁组,分别提取3组细胞的外泌体,将其孵育肝细胞系HepG2细胞后,检测肝细胞上清中谷丙转氨酶(ALT)、乳酸脱氢酶(LDH)水平,肝细胞存活率,以及肝细胞肿瘤坏死因子-α(TNF-α)与白细胞介素-6(IL-6)mRNA水平,评估外泌体对肝细胞的作用;采用RT-qPCR检测单核细胞来源外泌体中miR-155含量的变化;通过数据库检索、双重荧光素酶报告基因检测及Western blotting实验分析miR-155的潜在靶点;观察热打击单核细胞外泌体对肝细胞miR-155的影响,以及使用乌司他丁或miR-155抑制剂处理对这些外泌体功能的影响。结果热打击单核细胞来源外泌体使肝细胞上清ALT及LDH水平、TNF-αmRNA及IL-6 mRNA水平明显增高(P<0.01),细胞存活率下降(P<0.05);热打击能增加单核细胞及其释放外泌体内含的miR-155水平;miR-155可能以细胞因子信号传导抑制分子1(SOCS1)作为靶基因发挥作用;采用热打击单核细胞外泌体孵育肝细胞可使后者miR-155水平增高,乌司他丁或miR-155抑制剂能降低热打击单核细胞外泌体miR-155水平并减少肝细胞TNF-α和IL-6 mRNA的表达,差异有统计学意义(P<0.05)。结论热打击单核细胞可上调肝细胞炎性因子导致的炎症损伤,这可能是通过释放外泌体miR-155至肝细胞并以SOCS1为潜在靶点实现的;乌司他丁可能缓解此条件下肝细胞内炎性因子mRNA的表达。Objective To observe the influence of exosomal miR-155 derived from monocytes stimulated by heat stress on the inflammatory response of hepatocytes.Methods According to different treatments,we termed THP-1 monocytes into the control group,the heat stress group,and the ulinasatin group.We then extracted the exosomes from each group.To study the function of exsome on hepatocytes,we incubated hepatocytes with these exosomes.We then tested the alanine aminotransferase and lactate dehydrogenase levels in the supernatant,measured cell viability,and detected the relative mRNA levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the hepatocytes.Then,the expression changes of exosomal miR-155 derived from monocytes were detected using RT-qPCR.Furthermore,the potential target of miR-155 was analyzed using database retrieval,dual-luciferase report,and Western blotting assay.Finally,the effect of monocyte exosomes on hepatocyte miR-155 and the treatment with ulinastatin or miR-155 inhibitor on hepatocyte injury were observed.Results When incubating hepatocytes with the exosomes derived from monocytes stimulated by heat stress,the levels of alanine aminotransferase and lactate dehydrogenase in the supernatant and the mRNA levels of TNF-αand IL-6 in hepatocytes were all significantly increased(P<0.01)with decreased cell survival rate(P<0.05).Heat stress increased the level of miR-155 contained in monocytes and their release of exosomes.Database retrieval,dual-luciferase reports,and Western blotting assay showed that miR-155 might target SOCS1.The exosomes from monocytes stimulated by heat stress increased miR-155 in liver cells.Ulinastatin or miR-155 inhibitor could reduce the exosomal miR-155 level and decrease TNF-αand IL-6 mRNA expression in liver cells.The difference was statistically significant(P<0.05).Conclusion The monocytes stimulated by heat stress increase the inflammatory response of hepatocytes,which may be associated with increased exosomal miR-155 using SOCS1 as a potential target.Ulinastatin ma

关 键 词：中暑 外泌体 单核细胞 乌司他丁 肝细胞 MIR-155 

分 类 号：R594.13[医药卫生—内科学]