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
作者:
作者单位:

中南大学湘雅三医院 烧伤整形科,湖南 长沙 410013

作者简介:

尹朝奇,中南大学湘雅三医院副主任医师,主要从事烧伤脓毒症及创面修复方面的研究。

通信作者:

尹朝奇,Email: yinchaoqi@163.com

基金项目:

湖南省科技厅技术创新计划基金资助项目(2017SK50125)。


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

Department of Burns and Plastic Surgery, the Third Xiangya Hospital, Central South University, Changsha 410013, China

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 音频文件
  • |
  • 视频文件
    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ADARl基因敲除小鼠(ADAR1ECKO)与野生型小鼠(ADAR1flox/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,6 h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ADARl 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,ADAR1ECKO小鼠死亡时间早于ADAR1flox/flox小鼠,存活率低于ADAR1flox/flox小鼠(均P<0.05);组织病理学观察显示,注射LPS 6 h后,ADAR1ECKO小鼠的肝损伤比ADAR1flox/flox小鼠更严重;细胞免疫荧光观察显示,注射LPS 6 h后,ADAR1ECKO小鼠LSECs中Cav-1与VE-cadherin的表达低于ADAR1flox/flox小鼠。正常野生型小鼠的LSECs转染ADARl 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。

    Abstract:

    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 ADARl knockout mice (ADAR1ECKO) and 20 wild-type mice (ADAR1flox/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 ADARl 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 ADAR1ECKO mice was earlier than that of ADAR1flox/flox mice, and the survival rate of ADAR1ECKO mice was lower than that of ADAR1flox/flox mice after LPS injection (both P<0.05); the results of histopathological showed that the liver injury in ADAR1ECKO was severe than that in ADAR1flox/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 ADAR1ECKO mice than those from ADAR1flox/flox mice. In LSECs from normal wild-type mice after ADARl siRNA transfection, the tube formation ability was decreased, and the expression of Cav-1 downstream protein VE-cadherin was down-regulated, while the expression of β-Catenin had no obvious change.Conclusion The down-regulation or functional deficiency ADAR1 can cause the aggravation of SRLI, and the mechanism is probably associated with its regulating the activity of the Cav-1/VE-cadherin pathway. Thus, activation of the ADAR1/Cav-1/VE-cadherin pathway is potentially an effective strategy for prevention and treatment of SRLI.

    图1 小鼠肝组织HE染色(×200) A:ADAR1ECKO小鼠;B:ADAR1flox/flox小鼠Fig.1 HE staining of the mouse liver tissues (×200) A: ADAR1ECKO mouse; B: ADAR1flox/flox mouse
    图2 免疫荧光法检测LPS处理后两组小鼠LSECs中Cav-1和VE-cadherin的表达Fig.2 Expressions of Cav-1 and VE-cadherin in the LSECs from mice of the two groups after LPS treatment by cellular immunofluorescence
    图3 内皮细胞成管实验 A:空白对照组;B:对照siRNA组;C:ADARl siRNA组Fig.3 Endothelial tube formation assay A: Blank control group; B: Control siRNA group; C: ADARl siRNA group
    图4 LSECs转染ADAR1 siRNA后Cav-1下游蛋白的表达Fig.4 Expressions of the downstream proteins of Cav-1 in LSECs after ADAR1 siRNA transfection
    参考文献
    相似文献
    引证文献
引用本文

尹朝奇,陶如意,王少华,陈佳,唐封杰,刘灿,刘岱松,陈舒悦,周建大,陈丰原. 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[J].中国普通外科杂志,2022,31(7):913-919.
DOI:10.7659/j. issn.1005-6947.2022.07.008

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2022-03-13
  • 最后修改日期:2022-06-10
  • 录用日期:
  • 在线发布日期: 2022-07-31