摘要
目的观察PPAR-γ激活对血管紧张素Ⅱ诱导的体外培养条件下大鼠血管平滑肌(VSMCs)的细胞外基质(ECM)释放及结缔组织生长因子(CTGF)表达的影响。方法原代培养大鼠VSMCs,用不同浓度PPAR-γ激动剂rosiglitzone和/或抑制剂GW9662、BADGE预处理后,加入0.1μmol/L血管紧张素Ⅱ(AngⅡ)刺激24h。比色法检测各组细胞培养液中羟脯氨酸含量,实时荧光定量RT-PCR方法检测各组三型胶原(ColⅢ)、层粘连蛋白(FN)及CTGFmRNA表达,Western blotting检测各组CTGF、PPAR-γ蛋白表达情况,基于ELISA的DNA-binding检测各组PPAR-γ活性。结果 AngⅡ可增加VSMCs PPAR-γ表达,但明显抑制其活性(P<0.05,vs.Control),PPAR-γ激动剂rosiglitazone可显著增加PPAR-γ蛋白表达及活化(P<0.05,vs.AngⅡ)。rosiglitazone可降低细胞培养液中羟脯氨酸含量,下调VSMCs中ColⅢ、FN、CTGF mRNA表达,抑制CTGF蛋白表达,而其他PPAR-γ激动剂(pioglitazone、15-d-PGJ2)均有相似作用,PPAR-γ拮抗剂GW9662及BADGE可拮抗这一作用。结论在VSMCs中,PPAR-γ激活可抑制AngⅡ诱导的CTGF表达,同时抑制ECM沉积。提示PPAR-γ可能在血管纤维化中起重要作用。
Objective To observe the effects of PPAR-γ activation on angiotensin(Ang)Ⅱ-induced extra cellular matrix(ECM) production and connective tissue growth factor(CTGF) expression in vitro in rat vascular smooth muscle cells(VSMCs).Methods Cultured rat VSMCs were pretreated with PPAR-γ activator rosiglitazone or/and PPAR-γ antagonist GW9662 or BADGE before treated with AngⅡ for 24 h.The content of hydroxyproline in the cell culture medium was measured by colorimetric assay.Real-time RT-PCR was used to detect collagen Ⅲ(ColⅢ),fibronectin(FN) and CTGF expression.Protein expressions of CTGF and PPAR-γ were measured by Western blotting.ELISA-based DNA-binding was used to detect PPAR-γ activation.Results AngⅡ treatment up-regulated PPAR-γ expression but significantly decreased its activity when compared with control group.Pretreatment of cells with rosiglitazone significantly increased PPAR-γ expression and activation in AngⅡ-stimulated VSMCs.Rosiglitazone decreased hydroxyproline level in cell culture medium,down-regulated ColⅢ,FN and CTGF expressions in VSMCs.Other PPAR-γ activators had the same effect as rosiglitazone,but PPAR-γ antagonist partly reduced this effect.Conclusion PPAR-γ activation may inhibit CTGF expression and ECM production in VSMCs.PPAR-γ might act as a therapeutic target for vascular fibrosis.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期5-10,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30900617
No.81070219)
教育部博士点基金新教师基金资助项目(No.2008.6981036)
陕西省科技攻关项目(No.2007K13-03(13))
国家重点研究发展基础计划(973)(No.2011CB503904)~~
