肝细胞外泌体抑制小鼠非酒精性脂肪肝脂质沉积  

Inhibition of liver lipid deposition by exosomes derived from hepatocytes in mice with nonalcoholic fatty liver

作　　者：武艳霜 王岩金 杨馥吉 严永敏 WU Yanshuang;WANG Yanjin;YANG Fuji;YAN Yongmin(School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013;Department of Liver Disease, Third Hospital of Zhenjiang, Zhenjiang Jiangsu 212005, China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学附属镇江三院肝病科,江苏镇江212005

出　　处：《江苏大学学报（医学版）》2022年第4期332-337,共6页Journal of Jiangsu University：Medicine Edition

基　　金：江苏省科技项目(BE2021690)。

摘　　要：目的:探讨人肝细胞系LO2细胞来源的外泌体(exosomes derived from hepatocytes,LO2-Ex)对高脂肪饮食诱导的非酒精性脂肪肝(nonalcoholic fatty liver diseases,NAFLD)小鼠肝脏脂质沉积的影响。方法:培养正常人肝细胞系LO2细胞,超速离心法提取LO2-Ex。Nanosight纳米颗粒分析系统测定外泌体的粒径和浓度,透射电镜观察外泌体形态,免疫印迹法检测外泌体表面标志物。高脂肪饮食喂养雄性C57BL/6小鼠10周建立NAFLD小鼠模型,第11周连续尾静脉注射LO2-Ex 4周后收集肝组织,HE染色观察肝脏组织结构,计算肝脏指数(肝脏质量/净体重),油红O染色检测小鼠肝脏脂质沉积程度,天狼星红染色观察肝组织胶原沉积情况。结果:LO2-Ex的粒径为120 nm左右,具有外泌体特征性的脂质双层结构,表达外泌体特异蛋白CD9和TSG101,不表达Calnexin。与模型组相比,LO2-Ex组小鼠肝脏表面红润、细腻、有光泽。LO2-Ex能够改善NAFLD小鼠肝组织结构,提高小鼠肝脏指数。HE染色、油红O染色和天狼星红染色显示LO2-Ex减轻了肝组织的空泡变性、脂肪沉积和胶原沉积。结论:LO2-Ex可以抑制肝组织脂肪沉积,延缓小鼠NAFLD进展。Objective:To investigate the effect of exosomes derived from hepatocytes(LO2-Ex)on liver lipid deposition in mice with nonalcoholic fatty liver induced by high-fat diet.Methods:Normal human hepatocyte line LO2 cells were cultured and LO2-Ex was extracted by ultracentrifugation.The particle size and concentration of exosomes were measured by nanosight nanoparticle analysis system,the morphology of exosomes was observed by transmission electron microscope,and the markers of exosomes were detected by Western blotting.C57BL/6 male mice were fed with high-fat diet for 10 weeks to establish the mice model of nonalcoholic fatty liver.From the 11th week,LO2-Ex was injected into the caudal vein continuously for 4 weeks,and the liver tissue was collected after 4 weeks,the liver tissue structure was observed by HE staining,the liver index(liver mass/net weight)was calculated,the lipid deposition in the liver was detected by oil red O staining,and the collagen deposition in the liver was observed by Sirius red staining.Results:LO2-Ex,a particle size of about 120 nm,has a lipid bilayer structure characteristic of exosomes.It expresses exosome specific proteins CD9 and TSG101,but does not express Calnexin.Compared with the model group,the liver surface of mice in LO2-Ex group were ruddy,delicate and shiny.LO2-Ex can reduce the weight of mice and improve the liver index of mice.HE staining,oil red O staining and Sirius red staining showed that LO2-Ex reduced vacuolar degeneration,fat deposition and collagen deposition in liver tissues.Conclusion:LO2-Ex can inhibit lipid deposition in liver tissues and inhibit the progression of nonalcoholic fatty liver in mice.

关 键 词：肝细胞 外泌体 非酒精性脂肪肝病 空泡变性 脂质沉积 胶原沉积 

分 类 号：R683.42[医药卫生—骨科学]