目的 探索Connexin43(Cx43)在幼年非酒精性脂肪性肝病(NAFLD)小鼠模型肝脏中的表达变化,及其与肝细胞脂质沉积、内质网应激、炎症浸润及氧化应激的关系,为进一步探索可能的干预靶点奠定基础。方法 采用4周龄C57BL/6J(n=24只)小鼠,随机分为两组,分别给予正常饮食和高脂饮食13周,建立正常对照及NAFLD模型。测量小鼠体重、肝重。收集小鼠血液和肝脏标本,检测血糖、血脂、胆固醇、甘油三酯。采用HE和油红O染色观察肝脏病变程度。应用免疫组织化学染色检测Cx43在小鼠肝脏组织的表达及定位,及其与脂质沉积相关指标CD36,炎症浸润及氧化应激相关指标 CD68、F4/80及内质网应激相关指标GRP78、pIRE1表达的相关性。结果 与正常对照组比较,NAFLD组小鼠体重、肝重、甘油三酯、胆固醇和血糖显著增高,差异有统计学意义(P<0.001)。与对照组小鼠相比,NAFLD 小鼠肝脏Cx43 表达上调,GRP78、IRE1、CD68、F4/80、CD36表达上调(P<0.01)。CD36(r=0.724)、CD68(r=0.544)、F4/80(r=0.648)、GRP78(r=0.575)、IRE1(r=0.658)与Cx43均呈显著正相关(P<0.05)。结论 Cx43在幼年NAFLD小鼠模型中表达上调,可能参与调控幼年NAFLD小鼠肝脏中脂质沉积、内质网应激、炎症浸润及氧化应激等病理变化过程。
Abstract
Objective To explore the change of Connexin43(Cx43) expression in the liver of young mouse model of nonalcoholic fatty liver disease (NAFLD),and to analyze its relationship with hepatocyte lipid deposition,endoplasmic reticulum stress,inflammatory infiltration and oxidative stress,in order to lay a foundation for further exploration of possible intervention targets. Methods A total of 24 4-week-old C57BL/6J mice were selected in to this study,and were randomly divided into two groups fed with normal diet and high-fat diet for 13 weeks,respectively.Then normal control and NAFLD model were established.The body weight and liver weight of mice were measured.Blood and liver samples were collected to detect blood glucose,blood lipids,cholesterol and triglycerides.The degree of liver lesion was observed by HE and oil red O staining.Immunohistochemical staining was used to detect the expression and localization of Cx43 in mouse liver tissue,and its correlation with lipid deposition related index CD36,inflammatory infiltration and oxidative stress related index CD68,F4/80 and endoplasmic reticulum stress related index GRP78,pIRE1 expression. Results Compared with the normal control group,the body weight,liver weight,triglyceride,cholesterol and blood glucose levels in NAFLD group were significantly higher (P<0.001).At the same time,it was found that compared with the control group,the expression of Cx43 in the liver of NAFLD mice was significantly up-regulated,and the expression of GRP78,IRE1,CD68,F4/80 and CD36 was also up-regulated(P<0.01).Correlation analysis showed that the expression of Cx43 was positively correlated with the expression levels of CD36 (r=0.724),CD68 (r=0.544),F4/80 (r=0.648),F4/80(r=0.575) and IRE1 (r=0.658). Conclusion Hepatic Cx43 expression is up-regulated in the young mouse model of NAFLD,which may be involved in the regulation of the pathological changes of lipid deposition,endoplasmic reticulum stress,inflammatory infiltration and oxidative stress in fatty liver.
关键词
非酒精性脂肪性肝病 /
Connexin43 肝脏脂质沉积 /
内质网应激 /
炎症浸润 /
氧化应激
Key words
nonalcoholic fatty liver disease /
connexin43 /
liver lipid deposition /
endoplasmic reticulum stress /
inflammatory infiltration /
oxidative stress
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基金
国家自然科学基金青年科学基金项目(81402694)
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