摘要
目的利用适于PPAR转录激活效应检测的细胞模型,分析系列化合物对PPARα及γ亚型的活化作用。方法将PPARα(或PPARγ)表达质粒、报告基因质粒和内参照质粒共转染HepG2细胞,并以仅转染报告基因质粒和内参照质粒的细胞作为对照。5个化合物(LTD-1,3,4,6,7)分别以不同浓度作用于转染细胞24h,然后裂解细胞,测定细胞内报告基因质粒的荧光素酶活性,后者表示化合物对PPARα(或PPARγ)的激活强度。结果细胞模型稳定可靠,能够灵敏反映PPAR的特异激活效应。甘氨酸类化合物的检测表明,LTD-4、LTD-6、LTD-7均具有激活PPARα和PPARγ的双重效应,LTD-1则仅能激活PPARγ,而LTD-3对PPARα和PPARγ均无明显的激活作用。结论所建细胞模型适于PPARα和PPARγ转录激活作用的分析,化合物LTD-4,6,7具有激活PPARα和PPARγ的双重作用,为作为双激动剂先导化合物的深入研发提供了依据。
Aim To establish a screening system based on the engineered cells for identifying the synthesized compounds, which activated both the peroxisome prolif- erators activated receptor （PPAR） α and γ subtypes. Methods When HepG2 cells reached the exponential growth period, pSGS-PPARα or pSGS-PPARγ, pGL3- PPRE and pRL-TK were co-transfected into HepG2 cell according to the manufacture＇s protocol. Meanwhile, the cells nontransfected with pSGS-PPARα or pSGS-PPARγ were served as the control. The cells were separately stimulated by five synthesized chemical Glycine compounds （LTD-1,3,4,6 and 7） at different concentrations for 24 hrs. Then the fluorescent poten-cies of the stimulated cells were measured with Veritas microplate luminometer. Results The screening sys- tem was reliable and sensitive to reflect the activation of PPARs. It was indicated that LTD-4, LTD-6 and LTD-7 could activate both PPARα and PPARγ, of which LTD-7 was the most potent, while LTD-1 could only activate PPARγ and LTD-3 could not activate PPARα or PPARγ. Conclusions Among the com- pounds, LTD-4,6,7 exerted their dual effect on PPARα and PPARγ transactivation. Furthermore, LTD-7 was the best candidate for PPARα and PPARγ dual agonist.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第6期787-791,共5页
Chinese Pharmacological Bulletin
基金
军事医学科学院创新启动基金资助项目(NoYC0216)
