绿原酸激动AMPK调节糖脂代谢的抗糖尿病作用研究  

Chlorogenic acid modulates glucose and lipid metabolisms via AMPK activation in HepG2 cells and shows its anti-hyperglycemic effect on streptozocin-induced diabetic mice

作　　者：姚宏纪 薛兆毅 王瑞琪 姜承祚 熊婧妍 夏芷萱 刘嫱[1] 刘启兵[1] 张勇[1] YAO Hong-ji;XUE Zhao-yi;WANG Rui-qi;JIANG Cheng-zuo;XIONG Jing-yan;XIA Zhi-xuan;LIU Qiang;LIU Qi-bing;ZHANG Yong(Department of Pharmacology,Hainan Medical University,Haikou 571199,China;Research Center of Pharmaceutical Engineering Technology,Harbin University of Commerce,Harbin 150076,China)

机构地区：[1]海南医学院药理教研室,海南海口571199 [2]哈尔滨商业大学制药工程技术研究中心,黑龙江哈尔滨150076

出　　处：《海南医学院学报》2022年第9期654-662,共9页Journal of Hainan Medical University

基　　金：海南省自然科学基金青年基金项目(819QN226);海南省高等学校科学研究资助项目(Hnky2019ZD-32);大学生创新创业训练计划项目(202011810010,X20201181058)。

摘　　要：目的:探讨绿原酸(chlorogenic acid,CGA)调节糖脂代谢的作用及潜在机制。方法:通过MTT法、糖消耗检测、甘油三酯测定的方法研究CGA对HepG2细胞毒性及糖脂代谢的影响。采用Western blot结合AMP激动的蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)特异性抑制剂阻断实验探究CGA调节糖脂代谢的潜在分子机制。采用高脂饲料联合链脲佐菌素诱导糖尿病合并糖脂代谢紊乱的小鼠模型,探究CGA体内降糖调脂的抗糖尿病活性,并通过口服糖耐量实验和胰岛素抵抗实验试验对糖耐量及胰岛素抵抗进行评估,以及肝组织HE染色观察其脂肪变性病理改变。结果:CGA在100μmol/L以内未见显著细胞毒性,其能够显著促进肝细胞HepG2的糖消耗水平,降低棕榈酸诱导的细胞内甘油三酯的水平。Western blot结果显示,CGA能够显著上调p-AMPKα和p-ACC水平,提示显著激活AMPK和ACC,且CGA对AMPK的激动作用可以被AMPK特异性抑制剂compound C所阻断,同时,compound C也能够阻断糖消耗和甘油三酯积累。动物实验结果表明,CGA还表现出显著的降糖调脂的抗糖尿病活性,具有保护肝功能,改善糖耐量和胰岛素抵抗的作用。结论:CGA能够通过激活AMPK发挥降糖调脂的抗糖尿病作用,是潜在可应用糖尿病治疗的候选药物。Objective:To explore the hypoglycemic effects of chlorogenic acid(CGA)in vitro and in vivo and its regulat⁃ing effect on glucose and lipid metabolism via AMPK activation.Methods:The cytotoxicity,glucose consumption and intracellu⁃lar triglyceride(TG)were detected by commercial kits.Western blot was used to detect the associated protein levels after CGA treatment,and the inhibiter blocking experiments were also done.In vivo experiment,the fasting blood-glucose,lipid metabo⁃lism,liver function,insulin resistance,glucose tolerance,and pathological change were assessed in streptozocin-induced diabetic mice.Results:CGA exhibited no cytotoxicity up to 100μmol/L.It significantly increased glucose consumption and reduced the palmitic acid-induced increase in intracellular TG level in HepG2 cells at 50μmol/L and 100μmol/L.CGA up-regulated the level of p-AMPK(Thr172)and p-ACC(Ser79)in dose-dependent manners in vitro and in vivo.The stimulating effect of CGA on AMPK was completely blocked by compound c(CC)on HepG2 cells.And the efficacies of CGA on glucose consumption and in⁃tracellular TG accumulation were also completely blocked by CC pretreatment.The CGA also exhibited potent anti-diabetic ef⁃fects with hypoglycemic activity,improving insulin resistance and glucose tolerance,regulating glucose and lipid metabolism and protecting the liver function in vivo.Conclusion:Our results suggest that CGA can regulate glucose and lipid metabolism via AMPK activation and exhibit potent anti-hyperglycemic effect in streptozocin-induced diabetic mice.It may be used as a potential effective anti-diabetes drug.

关 键 词：绿原酸 糖脂代谢 抗糖尿病活性 AMPK 

分 类 号：R587.1[医药卫生—内分泌] R589.2[医药卫生—内科学]