黄芪多糖活化AMPK减轻游离脂肪酸对C2C12成肌细胞的细胞毒性  

Astragalus polysaccharide attenuates free fatty acid-induced toxicity by activating AMPK in C2C12 myoblasts

作　　者：宋杰[1] 李静[2] 胡阳黔[1] 刘坚[3] 欧阳静萍[3] 

机构地区：[1]湖北医药学院附属东风医院,湖北十堰442000 [2]湖北医药学院,湖北十堰442000 [3]武汉大学医学院,湖北武汉430079

出　　处：《中国病理生理杂志》2012年第2期298-301,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30370673;No.30771023)

摘　　要：目的:探讨黄芪多糖(APS)减轻游离脂肪酸(free fatty acids,FFAs)对骨骼肌细胞的毒性作用及其机制。方法:培养C2C12成肌细胞分5组:对照组、APS组、5-氨基咪唑-4-甲酰胺-1-β-D-呋喃核糖苷(5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside,AICAR)组、FFAs组和FFAs+APS组。MTT法检测C2C12细胞存活率;透射电镜观察细胞超微结构;Western blotting检测细胞内腺苷酸活化蛋白激酶(AMP-activatedprotein kinase,AMPK)、磷酸化腺苷酸活化蛋白激酶(p-AMPK)和磷酸化乙酰辅酶A羧化酶(p-ACC)表达水平;高效液相色谱法测定细胞胞浆AMP/ATP比值。结果:(1)与FFAs组比较,APS显著提高细胞存活率(P<0.05);(2)透射电镜观察发现,APS作用24 h后可以减轻FFAs导致的线粒体肿胀;(3)Western blotting结果显示:与FFAs组比较,APS作用24 h后显著增加p-AMPK表达(P<0.01)及p-ACC表达(P<0.05),而不影响总AMPK表达(P>0.05);(4)AMP/ATP比值检测结果:与FFAs组比较,APS作用24 h后细胞内AMP/ATP比值增加(P<0.01)。结论:APS可以减轻FFAs对骨骼肌细胞的毒性,其机制可能与保护线粒体、激活AMPK途径有关。AIM： To examine the effects of Astragalus polysaccharide（APS） on the toxicity of free fatty acids（FFAs） in C2C12 myoblasts.METHODS： C2C12 cells were randomly divided into 5 groups： control group,APS group,5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside（AICAR）group,FFAs group and FFAs＋APS group.MTT assay was used to observe the viability of C2C12 cells.C2C12 cells pretreated with FFAs at concentration of 0.25 mmol/L for 24 h were exposed to APS at dose of 200 mg/L for 24 h.The expression of total AMP-activated protein kinase（AMPK）,phosphorylated AMPK（p-AMPK） and phosphorylated acetyl-CoA carboxylase（p-ACC） was examined by Western blotting.The content of AMP and ATP was determined by HPLC.The structural changes of the mitochondria were examined by transmission electron microscopy.RESULTS： The results of MTT assay indicated that APS improved the viability of C2C12 cells pretreated with FFAs.In FFAs＋APS group,the ratio of AMP/ATP was increased after treatment with APS.No difference of total AMPK expression in C2C12 cells between APS group and FFAs group was observed.However,the expression of p-AMPK and p-ACC increased in APS group as compared with FFAs group.The results of transmission electron microscopy indicated that APS improved the vacuolar degeneration of mitochondria resulted from treatment with FFAs in C2C12 cells.CONCLUSION： In C2C12 cells,APS attenuates FFA-induced lipotoxity via a mitochondria-and AMPK-dependent mechanism.

关 键 词：黄芪多糖 游离脂肪酸 AMP激活蛋白激酶 乙酰辅酶A羧化酶 线粒体 

分 类 号：R363[医药卫生—病理学]