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。
Objective To investigate the role of hepatic stellate cell(HSC)inflammation in the pathogenesis of acute-on-chronic liver failure(ACLF).Methods A total of 45 male Kunming mice were randomly divided into control group,model group,and N-acetylcysteine(NAC)group.The mice in the model group and the NAC group were given injection of human serum albumin to establish a model of chronic liver disease,followed by intraperitoneal injection of the endotoxins lipopolysaccharide(LPS)and D-galactosamine(D-GlaN)to induce ACLF,and those in the control group were given injection of an equal volume of normal saline;the mice in the NAC group were given NAC since 1 week before the induction of NAC.The mice in the model group and the NAC group were sacrificed at 48 hours after the injection of LPS and D-GlaN.ELISA was used to measure the serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)and the levels of malondialdehyde(MDA)and superoxide dismutase(SOD)in liver tissue;HE staining was used to determine liver pathological score;ELISA was used to measure the serum levels of LPS and interleukin-1β(IL-1β).LX2 cells were stimulated by LPS and H2O2 with the presence or absence of NAC,and ELISA was used to measure the levels of IL-1βand interleukin-6(IL-6)in medium.LX2 cells were stimulated by LPS and H2O2,and then HL7702 cells were cultured with LX2 medium;Western blot was used to measure the expression of caspase-3 and caspase-8 in HL7702 cells,and flow cytometry was used to measure the apoptosis of HL7702 cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups;the least significant difference t-test was used for comparison of data with homogeneity of variance between two groups,and the Tamhane’s T2 test was used for comparison of data with heterogeneity of variance.The Kaplan-Meier survival analysis was used to evaluate survival time,and the log-rank test was used for comparison.Results At 48 hours,all mice in control group survived,while 3 mice in the mode
作者
田臻
王丽莎
姚耐娟
赵英仁
阮骊韬
TIAN Zhen;WANG Lisha;YAO Naijuan;ZHAO Yingren;RUAN Litao(Department of Infectious Diseases,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Department of Ultrasound,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Department of Obstetrics and Gynecology,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2021年第3期642-647,共6页
Journal of Clinical Hepatology
基金
国家自然科学基金(81800548)。
关键词
慢加急性肝功能衰竭
肝星状细胞
炎症
细胞凋亡
Acute-On-Chronic Liver Failure
Hepatic Stellate Cells
Inflammation
Apoptosis
