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

SONG Jie1,LI Jing2,HU Yang-qian1,LIU Jian3,OUYANG Jing-ping3 (1Dongfeng Affiliated Hospital of Hubei University of Medicine,2Hubei University of Medicine,Shiyan 442000,China;3Medical School of Wuhan University,Wuhan 430079,China.)  
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.
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