活性跟踪法研究地骨皮中的PPARγ激动作用成分  

Study on the active component PPARγ in Cortex lycii by the method of active tracking

作　　者：谢练武[1,2] 李顺祥[1] 潘宇[1] 喻嵘[1] 成细华[1] 

机构地区：[1]湖南中医药大学,长沙410208 [2]中南林业科技大学理学院,长沙410004

出　　处：《中华中医药杂志》2014年第8期2484-2488,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金(No.81273753;No.81102593);中国博士后科学基金(No.2013M531787);湖南省科技计划项目(No.2013RS4035);湖南省中药学重点学科资助(No.湘教通[2011]76号);中药有毒物质防控技术湖南省工程实验室项目(No.201304)~~

摘　　要：目的:筛选地骨皮中的过氧化物酶体增殖物激活受体γ(PPARγ)激动剂,确定具有调节糖脂代谢作用的潜在活性成分。方法:运用PPARγ受体表达质粒、报告质粒(tk-PPREx3-luc)与编码绿色荧光蛋白(pEGFP-N1)质粒短时转染HEK293细胞的体外筛选模型,通过活性跟踪分离筛选地骨皮中的PPARγ激动剂,评价配体浓度对PPARγ活性的响应。采用与高选择性PPARγ拮抗剂T0070907共孵育培养,分析PPARγ对配体的依耐程度。结果:地骨皮中的脂肪酸类具有较强的PPARγ激动活性,而芳香酸则无明显效果。脂肪酸的活性大小为:棕榈酸>亚麻酸>亚油酸>硬脂酸>油酸>蜂花酸。棕榈酸在1.0-10μg/mL范围内,浓度与PPARγ激动活性呈线性相关。与呲格列酮相比,T0070907能以相同模式阻断棕榈酸对PPARγ的激活作用。结论:地骨皮中具有PPARγ激动作用的主要组分是以棕榈酸为主的脂肪酸类。棕榈酸与呲格列酮类似,对PPARγ的激活作用也与浓度相关,而且PPARγ对棕榈酸也呈配体依耐性。Objective： To screen the active component PPARγ in Cortex lycii to make sure potential active ingredients that can regulate the lipid metabolism. Methods： The human HEK-293 ceils were transfected by PPARy receptor expression, PPAR luciferase reporter plasmid （tk-PPREx3-luc） and encoding enhanced green fluorescent protein plasmids （pEGFP-N1） that were used as screening model in vitro and be hatched together with the high selectivity PPARγ, antagonist T0070907. The dependence on ligand of PPARy was analyzed. Results： The fatty acids in Cortex lycii had strong PPARy agonist, while aromatic acid had no obvious effect. The activity of fatty acids was, in order, palmitic acid, linolenic acid, linoleic acid, stearic acid, oleic acid and melissic acid. The concentration of palmitic acid in 1.0-10μg/mL had linear relationship with the activity of PPARy antagonist T0070907, which could interrupt palmitic acid to active the PPARy by following the same mode as pioglitazone. Conclusion： The main active component PPARy in Cortex lycii is the fatty acids that mainly contained palmitic acid. The activation of PPARy by palmitic acid is related to its concentration and the PPARy showed ligand-dependency on palmitic acid, which is similar to the effect of pioglitazone on PPARγ.

关 键 词：地骨皮 过氧化物酶体增殖物激活受体Γ 活性跟踪分离 脂肪酸 棕榈酸 

分 类 号：R284[医药卫生—中药学]