基础医学与临床2017，Vol.37Issue(5)：619-624,6.
以PPARγ2为靶点的传统中药细胞学筛选方法的建立和应用
Establishment and application ofChinese herbal cytological screening method targeting at PPARγ2
摘要
Abstract
Objective To establish a natural Chinese herbal cytological screening method through the establishment of stably transfected 3T3-L1 cell line with the luciferase reporter gene expression plasmid containing human PPARγ2 gene promoter.Methods The luciferase reporter gene expression plasmids containing various lengthPPARγ2 gene promoter sequence were constructed by using pGL3-Basic-Luc and pGL3-Enhancer-Luc as vector.The stably transfected 3T3-L1 cell line was stablished, and twenty-eight kinds of extracts of Chinese medical herbs (ECMH) were selected.Finally the effects of these ECMH on the luciferase expression in 3T3-L1 cells were observed.Results The luciferase expression of 3T3-L1 cells stably transfected with pGL3-Enhancer-PPARγ2 625 bp-Luc plasmid was the highest.Therefore, this cell line was selected for the following screening experiment.Our results showed that the administration of safflower ECMH(10,100, and 1 000 μg/mL)significantly promoted the luciferase expression of 3T3-L1 cells.it was 1.63, 1.90, and 2.30(P<0.05) of no drug intervention cells.Similarity, Leaves and madder ECMH (10~1 000 μg/mL)also notably increased the luciferase expression of 3T3-L1 cells.The highest actions were observed, at a concentration of 1 000 μg/mL,which were 2.03(P<0.01) and 2.00 times of the control group(P<0.01), respectively.Conclusions Using this cell line safflower, leaves, madder ECMH were found to significantly promote human PPARγ2 gene promoter activities with the screening method, and they may become potential weight loss medicine in the future.关键词
过氧化物酶体增生物激活受体γ2/中药水溶性提取物/质粒构建/稳定转染/3T3-L1前脂肪细胞Key words
peroxisome proliferator activated receptor gamma/extracts of chinese medical herbs/plasmid construction/stable transfection/3T3-L1 preadipocytes分类
医药卫生引用本文复制引用
王欣,徐剑,朱惠娟,李乃适,王林杰,阳洪波,龚凤英,潘慧..以PPARγ2为靶点的传统中药细胞学筛选方法的建立和应用[J].基础医学与临床,2017,37(5):619-624,6.基金项目
国家自然科学基金(30540036,30771026,81370898) （30540036,30771026,81370898）
北京市自然科学基金(7082079,7122146) （7082079,7122146）
国家临床重点专科建设项目(WBYZ2011-873) （WBYZ2011-873）
北京协和医院基金(PUMCH 2013-020) （PUMCH 2013-020）
