腺病毒介导的小鼠过氧化物酶体增殖剂活化受体γ基因过度表达对Raw264.7细胞小窝蛋白-1基因和蛋白表达影响的实验研究  

PPARγ1 overexpression on caveolin-1 expression of Raw264.7 cells

作　　者：胡琴[1] 张运[1] 刘春喜[1] 张梅[1] 井玛[1] 贺红[1] 冯进波[1] 王蓉[1] 蒋桂花[1] 张宪军[1] 姜虹[1] 朱清[1] 

机构地区：[1]教育部和卫生部心血管重构和功能研究重点实验室山东大学齐鲁医院心内科

出　　处：《中华心血管病杂志》2006年第5期458-463,共6页Chinese Journal of Cardiology

摘　　要：目的比较腺病毒介导的小鼠过氧化物酶体增殖剂活化受体γ(PPARγ)基因过度表达与配体活化对小鼠Raw264.7巨噬细胞小窝蛋白-1(CAV-1)基因和蛋白表达的影响,探讨PPARγ基因对巨噬细胞CAV-1的调控作用及机制。方法首先构建表达小鼠PPARγ1基因的复制缺陷型腺病毒表达载体;将Raw264.7细胞随机分成对照组、PPARγ基因过度表达组、PPARγ活化剂曲格列酮干预组以及PPARγ基因过度表达和曲格列酮共刺激组进行干预;观察各组Raw264.7细胞PPARγ和CAV-1基因和蛋白的表达变化。结果RT-PCR检测对照组Raw264.7细胞有CAV-1基因的表达,免疫印迹法未发现CAV-1蛋白表达,但免疫细胞化学证实胞膜和核膜上均有少量CAV-1表达;经RT-PCR、免疫细胞化学和免疫印迹检测发现PPARγ基因过度表达组、曲格列酮干预组和二者共刺激组Raw264.7细胞CAV-1基因和蛋白表达明显增加(且共刺激组>PPARγ基因过度表达组>曲格列酮干预组,P<0.05)。对照组Raw264.7胞浆内PPARγ表达量较低,而PPARγ基因过度表达组和共刺激组PPARγ表达均明显增加(P<0.05),而曲格列酮干预组无明显变化。结论PPARγ基因活化或过度表达均能上调Raw264.7细胞CAV-1基因和蛋白表达。曲格列酮活化PPARγ基因,增加CAV-1基因和蛋白表达,但不增加PPARγ基因和蛋白表达水平。与单一作用比较,同时活化和过度表达PPARγ基因对CAV-1基因和蛋白的表达有累积效应,说明PPARγ的这一作用在配体活化下增强。推测曲格列酮上调CAV-1表达依赖于PPARγ,非本身药理特性所致。Objective To investigate the effect of PPARγ1 gene overexpression on caveolin-1 mRNA and protein expressions in a murine macrophage cell line Raw264.7. Methods Replication-deficient recombinant adenovirus expression vector of PPARγ1 was constructed using the AdEasy system. Raw264.7 cells were randomly treated as follows： P group （PPARγ1 gene overexpression）, T group （Troglitazone 40 μmol/L in DMSO）, PT group （PPARγ1 gene overexpression and Troglitazone） and control group. Changes of PPARγ1 and caveolin-1 at mRNA and protein levels were investigated. Results Caveolin-1 expression can be detected by RT-PCR in Raw264. 7, by immunocytochemistry method in cell and nuclear membrane but not by immunoblotting at protein level. Caveolin-1 expression at mRNA and protein levels in Raw264.7 were significantly higher in P, T and PT groups compared to control group and the expression was also significantly higher in PT group than that in P group and T group （ P 〈 0.05 ）. PPARγ expression was significantly increased in PT group and P group where remained unchanged in T group compared to control group. Conclusion PPARγ1 overexpression can upregulate caveolin-1 expression in macrophages. Troglitazone upregulated caveolin-1 expression in the bsence of increased PPAR,γ1 expressions at mRNA and protein levels.

关 键 词：动脉硬化 细胞质膜微囊蛋白 巨噬细胞 过氧化物酶体增殖剂活化受体γ 

分 类 号：R543.5[医药卫生—心血管疾病]