摘要
目的：探讨在大鼠肝硬化发病过程中,巨噬细胞的极化状态及其与肠源性内毒素血症-内质网应激反应之间的关系。方法：36只雄性SD大鼠随机分为正常对照组和肝硬化模型组。各组动物分别于造模第4周末、6周末和8周末处死取材。ELISA法检测大鼠血浆中ALT、内毒素、Hcy的水平和肝组织匀浆中i NOS、TNF-α、IL-6、Arg-1、IL-10的水平;肝组织切片行HE染色和VG染色;实时荧光定量PCR法检测大鼠肝组织中Grp78、NF-κB、IRF5、CD86、CD206和TGF-β1的mRNA表达。结果：与相应的正常对照组相比,肝硬化模型组动物血浆中ALT、内毒素、Hcy水平和肝组织中Grp78的mRNA表达在4周、6周和8周均随病程进展逐渐升高（P〈0.05）;肝组织中NF-κB、IRF5和CD86的mRNA表达以及i NOS、TNF-α、IL-6的水平均显著升高（P〈0.05）,且它们的变化趋势为6周水平最高,4周次之,8周最低;肝组织中CD206和TGF-β1的mRNA表达以及Arg-1、IL-10的水平在4周未见明显变化,在6~8周逐渐升高（P〈0.05）。相关性分析结果显示,血浆中内毒素水平与肝组织中Grp78的mRNA表达显著相关（P〈0.01）;血浆中内毒素水平和肝组织中Grp78的mRNA表达均与肝组织中CD86和CD206的mRNA表达显著相关（P〈0.01）。结论：肝损伤-肠源性内毒素血症-内质网应激-巨噬细胞极化途径可能是肝纤维化乃至肝硬化发病的重要机制。
AIM： To explore the state of macrophage polarization and its relation with intestinal endotoxemiaendoplasmic reticulum stress in the development of liver cirrhosis induced by multiple pathogenic factors in rats.METHODS： The male SD rats（ n = 36） were randomly divided into normal control group and liver cirrhosis model group,and sacrificed at the end of the 4th,6th and 8th weeks. The rat model of liver cirrhosis was induced by multiple pathogenic factors. The levels of alanine aminotransferase（ ALT）,endotoxin,homocysteine（ Hcy） in the plasma,and inducible nitric oxide synthase（ i NOS）,tumor necrosis factor-α（ TNF-α）,interleukin-6（ IL-6）,arginase-1（ Arg-1） and interleukin-10（ IL-10） in the liver tissues were detected by ELISA. Histopathological change of the liver was observed under microscope with the staining of hematoxylin and eosin（ HE） and van Gieson（ VG）. The expression of glucose-regulated protein78（ Grp78）,nuclear factor-kappa B（ NF-κB）,interferon-regulatory factor 5（ IRF5）,CD86,CD206 and transforming growth factor-β1（ TGF-β1） at mRNA levels in the liver tissues were detected by the method of real-time fluorescence quantitative PCR. RESULTS： Compared with the corresponding normal control group,the levels of ALT,endotoxin,Hcy in the plasma and Grp78 mRNA in the liver tissues in liver cirrhosis model group were significantly and gradually increased（ P〈0. 05）. The mRNA expression of NF-κB,IRF5 and CD86,and the protein levels of i NOS,TNF-α and IL-6 in the liver tissues were significantly increased（ P〈0. 05）,and they successively increased from the 4th week to the 6th week and decreased reversely at the 8th week. The mRNA expression of CD206,TGF-β1,Arg-1 and IL-10 in the liver tissues were significantly increased from the 6th week to the 8th week（ P〈0. 05）,and no significant difference at the 4th week was observed. The level of endotoxin in the plasma was correlated with the mRNA expression of Grp78 in the liver tissues（ P〈0. 01）. Both endotoxin in the plasma and Grp78 mRNA in the liver tissues were correlated with the mRNA expression of CD86 and CD206 in the liver tissues（ P〈0. 01）. CONCLUSION： The pathway of liver damage-intestinal endotoxemiaendoplasmic reticulum stress-macrophage polarization may be critical in the pathogenesis of liver cirrhosis induced by multiple pathogenic factors.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第5期880-885,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81070339)
山西省国际科技合作计划(No.2010081068)
山西省回国留学人员科研基金资助项目(No.211-091)
山西医科大学细胞生理学省部共建教育部重点实验室主任基金资助项目(No.2010-09)
长治医学院博士科研启动经费项目(No.2010-01)
关键词
肝硬化
肝纤维化
巨噬细胞
内质网应激
内毒素
Liver cirrhosis
Liver fibrosis
Macrophage
Endoplasmic reticulum stress
Endotoxin
