RNA特异性腺苷脱氨酶1与Caveolin 1在脓毒症相关肝损伤中的作用及机制  

Roles of adenosine deaminase RNA-specific adenosine deaminase 1 and Caveolin-1 in sepsis-related liver injury and the mechanism

作　　者：尹朝奇[1] 陶如意[1] 王少华[1] 陈佳[1] 唐封杰 刘灿[1] 刘岱松 陈舒悦 周建大[1] 陈丰原[1] YIN Chaoqi;TAO Ruyi;WANG Shaohua;CHEN Jia;TANG Fengjie;LIU Can;LIU Daisong;CHEN Shuyue;ZHOU Jianda;CHEN Fengyuan(Department of Burns and Plastic Surgery,the Third Xiangya Hospital,Central South University,Changsha 410013,China)

机构地区：[1]中南大学湘雅三医院烧伤整形科,湖南长沙410013

出　　处：《中国普通外科杂志》2022年第7期913-919,共7页China Journal of General Surgery

基　　金：湖南省科技厅技术创新计划基金资助项目(2017SK50125)。

摘　　要：背景与目的:脓毒症相关肝损伤(SRLI)发病机制尚不清楚,细菌内毒素(LPS)对肝脏血管内皮细胞的炎症损害可能是重要环节。前期研究提示,RNA特异性腺苷脱氨酶1 (ADAR1)可能通过调控内皮细胞功能相关蛋白Caveolin 1 (Cav-1)在参与血管内皮应激中的局部和全身炎症反应。因此,本研究初步探讨ADAR1与Cav-1在SRLI中的作用,以期为SRLI的早期防治寻找新的方法。方法:取ADAR1基因敲除小鼠(ADAR1^(ECKO))与野生型小鼠(ADAR1^(flox/flox))各20只,腹腔注射LPS(20 mg/kg)诱导脓毒症小鼠模型脓毒症模型,6h后每组小鼠各取10只,获取肝脏组织,并分离肝窦内皮细胞(LSECs),通过HE染色观察其肝脏病理学改变,用细胞免疫荧光法观察LSECs中Cav-1及其下游蛋白VE-cadherin的表达,两组其余小鼠用于生存率分析;用ADAR1 siRNA转染正常野生型小鼠的LSECs后,通过内皮细胞成管实验观察转染后LSECs的增殖情况、Western blot检测Cav-1下游相关蛋白的表达。结果:生存观察结果显示,注射LPS后,ADAR1^(ECKO)小鼠死亡时间早于ADAR1^(flox/flox)小鼠,存活率低于ADAR1^(flox/flox)小鼠(均P<0.05);组织病理学观察显示,注射LPS 6 h后,ADAR1^(ECKO)小鼠的肝损伤比ADAR1^(flox/flox)小鼠更严重;细胞免疫荧光观察显示,注射LPS 6 h后,ADAR1^(ECKO)小鼠LSECs中Cav-1与VEcadherin的表达低于ADAR1^(flox/flox)小鼠。正常野生型小鼠的LSECs转染ADAR1 siRNA后,成管能力明显减弱,Cav-1下游蛋白VE-cadherin的表达下调,但β-Catenin的表达无明显变化。结论:ADAR1的下调或功能缺失会导致SRLI加重,机制可能涉及其调控Cav-1/VE-cadherin通路的活性。因此,激活ADAR1/Cav-1/VE-cadherin通路可能是防治SRLI的有效策略。Background and Aims:The pathogenesis of sepsis-related liver injury(SRLI) remains unclear,and the inflammatory damage of bacterial lipopolysaccharides(LPS) to the hepatic endothelial cells may be an important process.Previous studies have suggested that RNA-specific adenosine deaminase 1(ADAR1)may be involved in local and systemic inflammatory responses during endothelial stress through regulating the endothelial cell function-related protein Caveolin-1(Cav-1).Therefore,this study was conducted as a preliminary assessment to analyze the roles of ADAR1 and Cav-1 in SRLI,so as to help find a new approach for early prevention and management of SRLI.Methods:Mouse sepsis models were induced in 20 ADAR1 knockout mice(ADAR1^(ECKO)) and 20 wildtype mice(ADAR1^(flox/flox)) by injection of LPS(20 mg/kg).Ten mice in each group were sacrificed at 6 h after LPS injection,the liver tissues were harvested for histopathological observation by HE staining and the liver sinusoidal endothelial cells(LSECs) were isolated for observation of the expressions of Cav-1 and its downstream protein VE-cadherin by cellular immunofluorescence.The remaining mice in the two groups were used for survival observation.In LSECs from normal wild-type mice after ADAR1 siRNA transfection,the proliferative ability was determined by endothelial tube formation assay,and the expressions of the relevant downstream proteins of Cav-1 were determined by Western blot analysis.Results:The results of survival observation showed that the time of death of ADAR1^(ECKO) mice was earlier than that of ADAR1^(flox/flox) mice,and the survival rate of ADAR1^(ECKO) mice was lower than that of ADAR1^(flox/flox) mice after LPS injection(both P<0.05);the results of histopathological showed that the liver injury in ADAR1^(ECKO) was severe than that in ADAR1^(flox/flox) mice at 6 h after LPS injection;the results of cellular immunofluorescence showed that the expressions of Cav-1 and VE-cadherin in LSECs were lower from ADAR1^(ECKO) mice than those from ADAR1^(flox/flox) mice.

关 键 词：脓毒症 肝损伤 内皮细胞 腺苷脱氨酶1 窖蛋白1 

分 类 号：R657.3[医药卫生—外科学]