天麻素通过激活AMPK通路减少油酸诱导的HL-7702细胞脂肪蓄积  

Gastrodin ameliorates oleic acid-induced fat accumulation through activation of AMPK pathway in HL-7702 cells

作　　者：耿雅娜[1] 于滨[1] 孔维佳[1] 

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050

出　　处：《中国药理学通报》2015年第1期39-44,共6页Chinese Pharmacological Bulletin

基　　金：国家科技部"十二五"重大新药创制科技重大专项(No2012ZX09301-002-001)

摘　　要：目的研究天麻素(GSTD)对油酸(OA)诱导的HL-7702细胞脂肪蓄积的抑制作用,并探讨其可能的细胞信号通路。方法以噻唑蓝(MTT)法测定GSTD对HL-7702细胞存活率的影响;以1 mmol·L-1的OA处理24 h诱导细胞脂肪变性,同时加入不同浓度的GSTD,油红O(ORO)染色观察细胞脂肪蓄积情况并测定细胞内三酰甘油(TG)含量;以Western blot检测GSTD处理后细胞中AMPKα和ACC的磷酸化水平;以compound C处理细胞,研究其对GSTD药效的影响。结果 GSTD浓度≤3 386.5μmol·L-1时对HL-7702细胞没有明显毒性。OA处理24 h后细胞中出现大量脂滴蓄积,TG含量明显增加,但同时加入浓度为169.3或338.7μmol·L-1的GSTD可明显抑制HL-7702细胞的脂肪蓄积,并使TG含量分别平均下降35%和43.6%(与单加OA的细胞比较P<0.01)。GSTD处理后能时间和浓度依赖性地增加细胞中的p-AMPKα和p-ACC水平,compound C能完全阻断GSTD对AMPK通路的激活作用以及减少肝细胞TG蓄积的作用。结论 GSTD可明显抑制OA诱导的HL-7702细胞脂肪蓄积,降低TG含量,其作用依赖于细胞中AMPK通路的激活。Aim To study the inhibitory effect of gast-rodin （GSTD） on oleic acid （OA）-induced fat accu-mulation in HL-7702 cells and explore possible cellular signaling pathways. Methods The MTT method was used to study the impact of GSTD on cell viability in HL-7702 cells. Cellular steatosis was induced by 1 mmol·L-1 of OA administration for 24 h, and differ-ent concentrations of GSTD were added at the same time. Oil red O （ ORO） staining was used to determine fat accumulation in cells, and intracellular triglyceride （ TG） contents were assayed. Western blot was used to determine the phosphorylation levels of AMPKα and ACC in cells after GSTD administration. Compound C was used to treat the cells in order to study its influ-ence on the efficacies of GSTD. Results GSTD had no obvious toxicity in HL-7702 cells when its concen-tration was≤3 386. 5 μmol · L^-1 . After 24 h of OA administration, there were large amounts of lipid drop-lets accumulated in HL-7702 cells, and intracellular TG contents greatly increased as well. However, when 169. 3 or 338. 7 μmol · L^-1 of GSTD was added to-gether with OA, fat accumulation in cells was greatly inhibited, and intracellular TG contents were reduced averagely by 35% and 43 . 6%, respectively （ P〈0. 01 vs OA alone ） . After administration, GSTD could in-crease the levels of p-AMPKα and p-ACC in HL-7702 cells time and dose dependently. Compound C could completely abolish the stimulating activity of GSTD on AMPK pathway and block its reducing effect on hepatic TG accumulation. Conclusions GSTD greatly inhibits OA-induced fat accumulation and reduces intracellular TG contents in HL-7702 cells;the efficacy of GSTD is dependent on the activation of cellular AMPK pathway.

关 键 词：天麻素 油酸 HL-7702 脂肪变性 三酰甘油 AMPK 

分 类 号：R284.1[医药卫生—中药学] R322.47[医药卫生—中医学]