摘要
目的:探讨PPARγ激活物对心脏非心肌细胞(cardiac nonmyocyte,CNM)增殖的影响及其可能涉及的PPARγ途径。方法:体外原代培养新生大鼠的CNM,以AngⅡ刺激,并分别给予不同浓度的吡格列酮和15-脱氧前列腺素J2(15d-PGJ2)。以MTT比色法和3H-胸腺嘧啶脱氧核苷掺入实验检测CNM增殖情况,以瞬时基因转染CNM测定PPARγ活化后对靶基因的转录调控活性。结果:AngII刺激后CNM明显增殖3、H-胸腺嘧啶脱氧核苷的掺入明显增加,经不同浓度的吡格列酮和15d-PGJ2作用后,呈剂量依赖性抑制CNM的增殖和3H-胸腺嘧啶脱氧核苷掺入。将PPARγ表达质粒载体与含乙酰辅酶A氧化酶(ACO)启动子的过氧化酶体增殖反应元件(PPRE)表达质粒共转染CNM,其荧光素酶的表达量较非共转染显著增强;以吡格列酮和15d-PGJ2刺激后,荧光素酶的相对表达量呈剂量依赖性显著增强。结论:PPARγ激活物吡格列酮和15d-PGJ2在体外显著抑制新生大鼠CNM的增殖,这一过程可能与PPARγ的激活有关。
Aim： To investigate the effect of PPARγ activators on inhibition of cardiac nonmyocytes （CNM） proliferation and the PPARγ- dependent pathway possibly involved. Methods： Angiotensin Ⅱ was used to induce proliferation of primarily cultured CNM from neonatal rats, and pioglitazone or 15-deoxy-△^12,14-prostaglandin J2 （15d-PGJ2） was applied to these CNM in various dosages in vitro. MTT assay and^3H-TdR uptake were determined to estimate proliferation of CNM, and transient transfection of reporter gene containing PPRE from ACO promoter （PPRE-pGL3） with or without PPARγ expression plasmid （PPARγ-pSG5） to CNM was performed to determine the control of target-gene transcription. Results： Angiotensin Ⅱ caused a significant increase in MTT value and ^3H-TdR uptake in CNM,which could be significantly reversed by pioglitazone and 15d-PGJ2 in a dose-dependent manner. Transient cotransfection of PPRE-pGL3 with PPARγ-pSG5 to CNM resulted in significant increase in luciferase activity compared with that without PPARγ-pSG5 cotransfection. Pioglitazone and 15d-PGJ2 induced increase in luciferase activity also in a dose-dependent manner. Conclusion： Pioglitazone and 15d-PGJ2, as the activators of PPARγ, inhibit proliferation of CNM from neonatal rats, the effect may be relate to the activation of PPARγ.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2006年第2期159-162,共4页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(30270551)
军队"十五"面上课题基金资助(02M012)
