地骨皮中酰胺类生物碱衍生物改善HepG2细胞脂质代谢的作用及机制研究  

Effect and Mechanism of Amide Alkaloids of Lycii Cortex on Improving Lipid Metabolism in HepG2 Cells

作　　者：侯颖 陈辉[1,2] 袁培培[1] 傅阳 郑晓珂[1,2] 冯卫生[1,2] HOU Ying;CHEN Hui;YUAN Pei-pei;FU Yang;ZHENG Xiao-ke;FENG Wei-sheng(School of Pharmacy,,Henan University of Chinese Medicine,Zhengzhou 450046,China;Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment&New Drug Research and Development of Henan Province,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,郑州450046 [2]河南中医药大学呼吸疾病诊疗与新药研发河南省协同创新中心,郑州450046

出　　处：《中国药学杂志》2020年第16期1339-1345,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(21602047);河南省高等学校青年骨干教师资助计划资助(2017GGJS-081)。

摘　　要：目的研究地骨皮中2个酰胺类生物碱衍生物(LCAA和LCAB)对HepG2细胞脂质代谢的改善作用,并探讨其可能的作用机制。方法建立油酸钠诱导的HepG2细胞脂质堆积模型,采用LCAA和LCAB干预细胞,油红O染色法观察细胞内脂质堆积情况。采用氧化酶法测定HepG2细胞中的TC含量,GPO-PAP法检测HepG2细胞中的TG含量;Western blot法检测脂代谢相关蛋白SIRT1、FOXO1、AMPK与phospho-AMPK(p-AMPK)蛋白表达。RT-qPCR法检测AMPK、SIRT1、ACC2、COXII和COXIII的mRNA表达。结果LCAA和LCAB均可显著减少油酸钠所致的HepG2细胞内脂质堆积(P<0.01),降低TC和TG水平(P<0.05或P<0.01)。同时油酸钠所致的p-AMPK蛋白表达量升高,SIRT1和FOXO1蛋白表达量降低(P<0.01),AMPK、SIRT1、ACC2、COXII和COXIII mRNA含量降低(P<0.05或P<0.01)均被LCAA和LCAB改善。结论LCAA和LCAB可显著改善HepG2细胞脂质代谢,并可通过调节脂代谢相关蛋白p-AMPK、SIRT1、FOXO1和线粒体氧化相关因子COXII、COXIII发挥调血脂作用。OBJECTIVE To investigate the effect of two amide alkaloid derivatives(LCAA and LCAB)from Lycii Cortex on lipid metabolism in HepG2 cells and explore their possible mechanisms.METHODS The lipid accumulation model of HepG2 cells induced by sodium oleate was established.The cells were pretreated with different concentrations of LCAA and LCAB,and the lipid accumulation was observed by oil red O staining.The content of TC in HepG2 cells was measured by oxidizing enzyme method,and the content of TG in HepG2 cells was measured by GPO-PAP method.The expression of lipid metabolism related proteins,such as SIRT1,FOXO1,AMPK and phospho-AMPK(p-AMPK),was detected by Western blot.The mRNA expression of AMPK,SIRT1,ACC2,COXII and COXIII were detected by RT-qPCR.RESULTS Both LCAA and LCAB could significantly decrease the lipid accumulation in HepG2 cells induced by sodium oleate(200μmol·L-1)(P<0.01)and the TC and TG levels(P<0.05 or P<0.01).At the same time,the sodium oleate-induced high expression of p-AMPK protein,the low expression of SIRT1,FOXO1(P<0.01),and the low expression of AMPK,SIRT1,ACC2,COXII and COXIII mRNA(P<005 or P<001)were all improved by LCAA and LCAB.CONCLUSION Both LCAA and LCAB can significantly improve lipid metabolism in HepG2 cells,and regulate lipid metabolism by regulating lipid metabolism-related proteins p-AMPK,SIRT1,FOXO1 and mitochondrial oxidation-related factors COXII and COXIII.

关 键 词：地骨皮 酰胺类生物碱衍生物 HEPG2细胞 脂质堆积 脂代谢 

分 类 号：R965[医药卫生—药理学]