摘要
目的:探讨非酒精性脂肪肝患者血清中exosomes的水平及其对肝细胞脂代谢的影响。方法:通过Western-blotting,透射电镜、Nanosight鉴定exosomes;并通过BCA蛋白定量的方法检测19例NAFLD患者(NAFLD组),18例健康对照(健康对照组)血清中exosomes内蛋白含量;将10μg的exosomes与HepG2细胞共培养,48 h后,收集细胞,GPO-PAP法检测其中总胆固醇(TC)和总甘油三酯(TG)的表达;通过Western-blotting检测腺苷酸活化蛋白激酶(AMPK)的磷酸化水平。结果:exosomes高表达CD9、CD63、CD81,呈圆形或椭圆形,直径在40～210(110.00±21.36) nm。NAFLD组中的exosomes蛋白水平明显高于健康对照(P<0.05)。exosomes与HepG2细胞共培养后,其TC、TG的表达水平与对照组相比明显上调,差异均有统计学意义(P<0.05)。结论:exosomes可影响AMPK通路的活性,在肝细胞的脂代谢过程中发挥了重要作用,促进了非酒精性脂肪肝的发展。
Objective:To investigate the level of serum exosomes in patients with nonalcoholic fatty liver disease(NAFLD)and its effect on lipid metabolism in hepatic cells.Method:Western-blotting,transmission electron microscope and Nanosight were used to identify the character of exosomes.The protein content in serum exosomes in 19 cases of NAFLD(NAFLD group)and 18 cases of healthy controls(healthy control group)were detected by BCA protein assay.10μg of exosomes were co-cultured with HepG2 cells.After 48 h,the cells were harvested,and the expression of total cholesterol(TC)and total triglyceride(TG)was detected by GPO-PAP method.The phosphorylation level of adenosine monophosphate activated protein kinase(AMPK)was detected by Western-blotting.Result:The expression of CD9,CD63 and CD81 were high in exosomes,which was round or elliptical with a diameter of 40-210(110.00±21.36)nm.The protein level of exosomes in NAFLD group was higher than that of healthy control group(P<0.05).After co-culture of exosomes with HepG2 cell,the expression of TC and TG significantly increased compared with those in control group(P<0.05).Conclusion:Exosomes can affect the activity of AMPK pathway and play an important role in the lipid metabolism of hepatocyte,which promote the development of NAFLD.
作者
石柳
汤建华
温建军
曹德清
刘雪
林晔
张静智
SHI Liu;TANG Jianhua;WEN Jianjun(Ganzhou People’s Hospital,Ganzhou 341000,China)
出处
《中国医学创新》
CAS
2018年第30期13-16,共4页
Medical Innovation of China
基金
江西省卫生计生委科技计划项目(20187226)
