天麻粉水浸出物抑制油酸诱导的HL-7702细胞三酰甘油蓄积的实验研究  

Experimental study of inhibitory effect of Gastrodia powder aqueous extract on oleic acid-induced triglyceride accumulation in HL-7702 cell

作　　者：左增艳[1] 于滨[1] 耿雅娜[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：《中国当代医药》2015年第13期12-15,21,共5页China Modern Medicine

基　　金：中央级公益性科研院所基本科研业务费专项项目(IMBF20060505)

摘　　要：目的研究天麻粉水浸出物抑制油酸(OA)诱导的HL-7702细胞三酰甘油(TG)蓄积的作用及相关细胞信号通路。方法以1 mmol/L的OA诱导细胞脂肪变性,同时加入不同浓度的天麻粉水浸出物共同孵育24 h后测定细胞内TG含量。在Western blot实验中,细胞以天麻粉水浸出物做相应处理后检测AMP活化蛋白激酶α(AMPKα)和乙酰辅酶A羧化酶(ACC)的磷酸化水平。在使用compound C的实验中,细胞以10μmol/L的compound C预处理30 min,然后加入天麻粉水浸出物,共同孵育8 h后进行Western blot实验;或加入OA+天麻粉水浸出物,共同孵育24 h后测定细胞内TG含量。结果 OA孵育24 h后,HL-7702细胞内TG含量较对照细胞升高约(4.0±0.5)倍(P<0.01),天麻粉水浸出物使细胞内TG呈剂量依赖性降低。天麻粉水浸出物处理后能剂量和时间依赖性地激活HL-7702细胞的AMPK信号通路,表现为磷酸化AMPKα(p-AMPKα)和磷酸化ACC(p-ACC)的水平显著增加。compound C预处理并与天麻粉水浸出物共同孵育后能完全阻断后者对AMPK通路的激活作用以及减少细胞内TG的药效。结论天麻粉水浸出物能显著抑制OA诱导的HL-7702细胞三酰甘油蓄积,此作用依赖于细胞内AMPK通路的激活。Objective To study the inhibitory effect of Gastrodia powder aqueous extract on oleic acid （OA）-induced triglyceride （TG）accumulation in HL-7702 cell and related cellular signaling pathway. Methods 1 mmol/L of OA was used to induce cellular steatosis,Gastrodia powder aqueous extract at different concentrations were added and incubated together for 24-h for determination of intracellular TG content.In Western blot experiment,cells were treated with Gas-trodia powder aqueous extract as indicated,phosphorylation level of AMP-activated protein kinaseα（AMPKα） as well as acetyl coA carboxylase（ACC）were determined.In experiment using compound C,cells were pretreated with 10 μmol/L of compound C for 30-min and then Gastrodia powder aqueous extract was added.After 8-h of incubation,Western blot was performed.Alternatively,the cells were treated with OA＋Gastrodia powder aqueous extract for 24 h after compound C pretreatment,intracellular TG contents were then assayed. Results Compared with that of control cell,the intracellular TG content increased for about （4.0±0.5） fold in HL-7702 cell after 24-h of OA incubation （P〈0.01）.Gastrodia powder aqueous extract could decrease intracellular TG in a dose-dependent manner.Administration of Gastrodia powder aque-ous extract could activate the AMPK pathway dose and time dependently in HL-7702 cell,as indicated by the obvious increase of phosphorylated AMPK α（p-AMPKα）as well as phosphorylated ACC（p-ACC）level.When the cells were pre-treated with compound C and incubated with Gastrodia powder aqueous extract,the stimulating activity of Gastrodia powder aqueous extract on AMPK pathway and its reducing effect on intracellular TG accumulation were completely blocked. Conclusion Gastrodia powder aqueous extract inhibits OA-induced TG accumulation greatly in HL-7702 cell, and this inhibitory effect depends on the activation of AMPK pathway in cells.

关 键 词：天麻粉 水浸出物 HL-7702细胞 油酸 三酰甘油 AMP活化蛋白激酶 

分 类 号：R965[医药卫生—药理学]