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肝星状细胞在慢加急性肝衰竭小鼠模型发病进程中的作用及机制

田臻 王丽莎 姚耐娟 赵英仁 阮骊韬

田臻, 王丽莎, 姚耐娟, 等. 肝星状细胞在慢加急性肝衰竭小鼠模型发病进程中的作用及机制[J]. 临床肝胆病杂志, 2021, 37(3): 642-647. DOI: 10.3969/j.issn.1001-5256.2021.03.027
引用本文: 田臻, 王丽莎, 姚耐娟, 等. 肝星状细胞在慢加急性肝衰竭小鼠模型发病进程中的作用及机制[J]. 临床肝胆病杂志, 2021, 37(3): 642-647. DOI: 10.3969/j.issn.1001-5256.2021.03.027
TIAN Z, WANG LS, YAO NJ, et al. Role and mechanism of hepatic stellate cells in the pathogenesis of mice with acute-on-chronic liver failure [J]. J Clin Hepatol, 2021, 37(3): 642-647. DOI: 10.3969/j.issn.1001-5256.2021.03.027
Citation: TIAN Z, WANG LS, YAO NJ, et al. Role and mechanism of hepatic stellate cells in the pathogenesis of mice with acute-on-chronic liver failure [J]. J Clin Hepatol, 2021, 37(3): 642-647. DOI: 10.3969/j.issn.1001-5256.2021.03.027

肝星状细胞在慢加急性肝衰竭小鼠模型发病进程中的作用及机制

DOI: 10.3969/j.issn.1001-5256.2021.03.027
基金项目: 

国家自然科学基金 81800548

详细信息
    作者简介:

    田臻(1990—),男,博士,主要从事肝脏疾病基础、临床及超声诊断研究

    通讯作者:

    田臻,tianzen1984@163.com

  • 利益冲突声明:本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突,特此声明。
  • 作者贡献说明:田臻负责课题设计、拟定写作思路;王丽莎、姚耐娟负责实验操作、论文撰写;田臻、王丽莎负责数据分析;田臻、赵英仁、阮骊涛负责论文审定。
  • 中图分类号: R575.3

Role and mechanism of hepatic stellate cells in the pathogenesis of mice with acute-on-chronic liver failure

  • 摘要:   目的  探究肝星状细胞(HSC)炎症在慢加急性肝衰竭(ACLF)小鼠模型发病过程中的作用及机制。  方法  45只雄性昆明种小鼠随机分为对照组、模型组及N-乙酰-L-半胱氨酸(NAC)组,每组15只。模型组、NAC组注射人血白蛋白构建慢性肝病模型,之后腹腔注射内毒素(LPS)+D-GlaN诱导ACLF,对照组注射等量生理盐水, NAC组诱导ACLF前1周开始应用NAC。模型组及NAC组小鼠腹腔注射LPS+D-GlaN 48 h后处死。ELISA法检测血清AST、ALT及肝组织丙二醛及超氧化物歧化酶水平,肝组织行HE染色并进行病理评分,ELISA法检测小鼠血清LPS、IL-1β水平。在有或无NAC的情况下用LPS、H2O2刺激LX2细胞,ELISA法检测培养基中IL-1β及IL-6水平。用LPS、H2O2刺激LX2细胞,收取LX2培养基培养HL7702细胞,Western Blot检测HL7702细胞Caspase 8和Caspase 3表达,流式细胞法检测细胞凋亡。计量资料多组间比较采用单因素方差分析,方差齐时用LSD-t检验进行两两比较,方差不齐时采用Tamhane’s T2进行检验。Kaplan-Meier法绘制生存曲线,生存分析采用log-rank检验。  结果  48 h时对照组小鼠全部存活,模型组小鼠存活3只、NAC组存活8只。NAC组小鼠48 h累积生存率优于模型组(P<0.001);模型组血清AST、ALT及肝组织丙二醛水平较对照组和NAC组显著升高,肝组织超氧化物歧化酶水平显著降低(P值均<0.001);模型组小鼠肝脏病理评分明显高于对照组和NAC组(P值均<0.05)。LPS、H2O2均可促进LX2细胞产生IL-1β、IL-6,NAC能够有效抑制LPS、H2O2的促炎作用(P值均<0.05);H2O2、LPS作用于LX2细胞促进HL7702细胞凋亡(P值均<0.05)。  结论  LPS通过ROS促进HSC炎症,诱导肝细胞凋亡参与肝衰竭的疾病进程。
  • 图  1  各组小鼠的生存曲线

    图  2  各组小鼠血清LPS和IL-1β水平比较

    图  3  各组小鼠肝组织HE染色(×200)

    注: a, 对照组;b, 模型组; c,NAC组。

    图  4  各组小鼠肝脏病理评分

    图  5  NAC抑制LPS诱导的LX2炎症

    图  6  共培养条件下LPS/H2O2可通过HSC促进肝细胞凋亡

    注:a~b, Western Blot检测HL7702细胞凋亡蛋白Caspase3和Caspase8的表达;c~d, Annexin V-PI染色后流式细胞术检测HL7702细胞的凋亡和坏死。

    表  1  各组小鼠血清ALT、AST及肝组织MDA和SOD水平

    组别 小鼠数(只) AST(U/L) ALT(U/L) MDA(mmol/g) SOD(U/mg)
    对照组 15 25.95±3.96 21.21±2.30 2529.21±143.32 152.16±17.13
    模型组 15 6184.62±1123.121)2) 5012.41±988.351)2) 9721.34±215.381)2) 51.67±5.591)2)
    NAC组 15 965.51±106.011) 845.32±115.751) 4231.79±157.051) 83.34±7.561)
    F 628.342 896.813 119.820 126.351
    P <0.001 <0.001 <0.001 <0.001
    注:与对照组相较,1)P<0.01;与NAC组相较,2)P<0.01。
    下载: 导出CSV
  • [1] Liver Failure and Artificial Liver Group, Chinese Society of Infectious Diseases, Chinese Medical Association; Severe Liver Disease and Artificial Liver Group, Chinese Society of Hepatology, Chinese Medical Association. Guideline for diagnosis and treatment of liver failure(2018)[J]. J Clin Hepatol, 2019, 35(1): 38-44. (in Chinese)

    中华医学会感染病学分会肝衰竭与人工肝学组, 中华医学会肝脏病学分会重型肝病与人工肝学组. 肝衰竭诊治指南(2018年版)[J]. 临床肝胆病杂志, 2019, 35(1): 38-44.
    [2] ZHANG J, ZHOU XM. Value of different scoring systems in predicting short-term mortality of patients with acute-on-chronic liver failure[J]. J Clin Hepatol, 2019, 35(9): 1990-1994. (in Chinese) DOI: 10.3969/j.issn.1001-5256.2019.09.021

    张静, 周新民. 5种评分系统对慢加急性肝衰竭患者短期病死率的预测价值[J]. 临床肝胆病杂志, 2019, 35(9): 1990-1994. DOI: 10.3969/j.issn.1001-5256.2019.09.021
    [3] MEHTA G, MOOKERJEE RP, SHARMA V, et al. Systemic inflammation is associated with increased intrahepatic resistance and mortality in alcohol-related acute-on-chronic liver failure[J]. Liver Int, 2015, 35(3): 724-34. DOI: 10.1111/liv.12559
    [4] PAN C, GU Y, ZHANG W, et al. Dynamic changes of lipopolysaccharide levels in different phases of acute on chronic hepatitis B liver failure[J]. PLoS One, 2012, 7(11): e49460. DOI: 10.1371/journal.pone.0049460
    [5] FUJITA T, SOONTRAPA K, ITO Y, et al. Hepatic stellate cells relay inflammation signaling from sinusoids to parenchyma in mouse models of immune-mediated hepatitis[J]. Hepatology, 2016, 63(4): 1325-1339. DOI: 10.1002/hep.28112
    [6] LI J, ZHAO YR, TIAN Z. Roles of hepatic stellate cells in acute liver failure: From the perspective of inflammation and fibrosis[J]. World J Hepatol, 2019, 11(5): 412-420. DOI: 10.4254/wjh.v11.i5.412
    [7] JIN L, GAO H, WANG J, et al. Role and regulation of autophagy and apoptosis by nitric oxide in hepatic stellate cells during acute liver failure[J]. Liver Int, 2017, 37(11): 1651-1659. DOI: 10.1111/liv.13476
    [8] TIAN Z, CHEN Y, YAO N, et al. Role of mitophagy regulation by ROS in hepatic stellate cells during acute liver failure[J]. Am J Physiol Gastrointest Liver Physiol, 2018, 315(3): g374-g384. DOI: 10.1152/ajpgi.00032.2018
    [9] DAS J, GHOSH J, MANNA P, et al. Acetaminophen induced acute liver failure via oxidative stress and JNK activation: Protective role of taurine by the suppression of cytochrome P450 2E1[J]. Free Radic Res, 2010, 44(3): 340-55. DOI: 10.3109/10715760903513017
    [10] MONIAUX N, DARNAUD M, GARBIN K, et al. The reg3α (HIP/PAP) lectin suppresses extracellular oxidative stress in a murine model of acute liver failure[J]. PLoS One, 2015, 10(5): e0125584. DOI: 10.1371/journal.pone.0125584
    [11] KINUGASA H, WHELAN KA, TANAKA K, et al. Mitochondrial SOD2 regulates epithelial-mesenchymal transition and cell populations defined by differential CD44 expression[J]. Oncogene, 2015, 34(41): 5229-39. DOI: 10.1038/onc.2014.449
    [12] KIM SR, KIM DI, KIM SH, et al. NLRP3 inflammasome activation by mitochondrial ROS in bronchial epithelial cells is required for allergic inflammation[J]. Cell Death Dis, 2014, 5: e1498. DOI: 10.1038/cddis.2014.460
    [13] GAO YD, TIAN Y, ZHANG XY, et al. Effect of magnesium isoglycyrrhizinate on concanavalin A-induced acute liver failure in mice[J]. J Clin Hepatol, 2020, 36(7): 1571-1576. (in Chinese) DOI: 10.3969/j.issn.1001-5256.2020.07.024

    高钰迪, 田原, 张向颖, 等. 异甘草酸镁对刀豆蛋白A诱导的急性肝衰竭小鼠模型的影响[J]. 临床肝胆病杂志, 2020, 36(7): 1571-1576. DOI: 10.3969/j.issn.1001-5256.2020.07.024
    [14] EDWARDS S, LALOR PF, NASH GB, et al. Lymphocyte traffic through sinusoidal endothelial cells is regulated by hepatocytes[J]. Hepatology, 2005, 41(3): 451-459. DOI: 10.1002/hep.20585
    [15] BIEGHS V, VERHEYEN F, van GORP PJ, et al. Internalization of modified lipids by CD36 and SR-A leads to hepatic inflammation and lysosomal cholesterol storage in Kupffer cells[J]. PLoS One, 2012, 7(3): e34378. DOI: 10.1371/journal.pone.0034378
    [16] DECHÂNE A, SOWA JP, GIESELER RK, et al. Acute liver failure is associated with elevated liver stiffness and hepatic stellate cell activation[J]. Hepatology, 2010, 52(3): 1008-1016. DOI: 10.1002/hep.23754
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出版历程
  • 收稿日期:  2020-08-19
  • 修回日期:  2020-09-28
  • 刊出日期:  2021-03-16
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