腺苷酸基琥珀酸活化AMPK抑制肝细胞内脂质蓄积的作用机制  

Adenylosuccinate, a metabolite in purine salvage pathway, binds to AMP-activated protein kinase(AMPK) γ subunit and enhances the metabolic efficiency of lipid

作　　者：王洋 姜允嘉 成钟 赵晓宏 郭鹏 蔡大勇 谢勇 WANG Yang;JIANG Yun-jia;CHENG Zhong;ZHAO Xiao-hong;GUO Peng;CAI Da-yong;XIE Yong(Department of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所中草药物质基础与资源利用教育部重点实验室,北京100193

出　　处：《中国医药生物技术》2020年第3期240-248,共9页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81473114)。

摘　　要：目的揭示腺苷酸基琥珀酸(盐)(S-AMP)的生物学新功能。方法以AMP活化蛋白激酶(AMPK)的天然底物AMP为参照,应用表面等离子共振研究S-AMP和AMPK的相互作用特征。分别构建脂质蓄积和糖蓄积的HepG2细胞模型,通过特定浓度的S-AMP和对照药物作用后测定Hep G2细胞内甘油三酯浓度和葡萄糖浓度,研究S-AMP和AMP降脂和降糖的作用规律。用Westernblot研究S-AMP作用HepG2细胞后提高AMPK磷酸化水平及其下游糖、脂质代谢通路中关键蛋白质的表达量变化规律,提出S-AMP的作用机制。最后利用饮食诱导的糖尿病和高血脂症金黄地鼠模型研究S-AMP促进糖脂代谢的药效。结果 S-AMP和AMPKγ亚基形成复合体后提高AMPK磷酸化水平,提高AMPK下游的甘油三酯脂肪酶和乙酰辅酶A羧化酶的表达量和磷酸化。促进葡萄糖分解的PFKFB3(6-phosphofructo-2-kinase/fructose-2,6-biphosphatase3 enzyme)蛋白和转化为糖原的蛋白质糖原合成酶的表达量没有明显变化,S-AMP显示出的降脂活性高于同等浓度的洛伐他汀。结论发现了S-AMP降低脂质蓄积的作用机制。Objective To discovery of novel biological functions of adenosylsuccinate(adenosylsuccinatic acid;referred to as S-AMP). Methods Surface plasmonic resonance(SPR) was used to explore the direct interaction between S-AMP and AMP activated protein kinase(AMPK) by using the natural substrate AMP of AMP-activated protein kinase(AMPK) as the reference. HepG2 cell models of lipid and glucose accumulation were constructed respectively to evaluate the concentration of triglyceride and glucose in HepG2 cells after the addition of S-AMP at different concentrations, the lipid-lowering and glucose-lowering effects of S-AMP were studied. Western blot was used to study the phosphorylation of AMPK and the expression of downstream signaling pathways of glucose and lipid metabolism by the addition of S-AMP to HepG2 cells, and the mechanism of S-AMP on the attenuation of lipid metabolism was proposed. Finally, the effect of S-AMP was verified in mice with diet-induced diabetes and hyperlipidemia. Results It was confirmed for the first time that the formation of complex of S-AMP and AMPK increased the phosphorylation level of AMPK, enhanced the expression of ATGL and ACC2 in the fatty acid synthesis downstream of AMPK pathway, and simultaneously increased the phosphorylation of ACC2. It also increased the intracellular decomposition of triglyceride to generate fatty acids and decreased the lipid accumulation in cells. In addition, the in vivo and in vitro experiments showed that S-AMP had no significant hypoglycemic activity. Conclusion The mechanism of S-AMP reducing lipid accumulation was confirmed.

关 键 词：腺苷酸基琥珀酸(盐) AMPK 脂质蓄积 作用机制 

分 类 号：R965[医药卫生—药理学]