腺病毒介导的mPPARγ1基因转染抑制IFN-γ诱导ECV304细胞galectin-9基因和蛋白表达  

Adenovirus-mediated mPPARγ1 gene overexpression inhibits IFN-γ-induced galectin-9 gene and protein expression in ECV304

作　　者：胡琴[1] 张宪军[2] 蒋桂花[1] 张运[1] 

机构地区：[1]山东大学齐鲁医院心内科,国家教育部和卫生部心血管重构和功能研究重点实验室,山东济南250012 [2]山东大学齐鲁医院皮肤科,山东济南250012

出　　处：《中国病理生理杂志》2008年第2期279-284,共6页Chinese Journal of Pathophysiology

摘　　要：目的:观察腺病毒介导的mPPARγ1转染抑制IFN-γ诱导ECV304细胞galectin-9基因和蛋白表达。方法:构建表达小鼠PPARγ1基因的复制缺陷型腺病毒表达载体;将融合80%的ECV304细胞给予不同刺激量(1×104U/L、5×104U/L、1×105U/L和2×105U/L)的IFN-γ干预;将IFN-γ(1×105U/L)预刺激并孵育24 h的ECV304细胞分成对照组(C)、PPARγ基因过度表达组(P)、PPARγ活化剂曲格列酮干预组(T)以及PPARγ基因过度表达和曲格列酮共刺激组(PT)进行干预,观察不同剂量IFN-γ对ECV304细胞galectin-9基因和蛋白表达的作用,以及PPARγ基因过度表达和/或活化对上述作用的影响。结果:正常ECV304细胞galectin-9基因表达弱。IFNγ孵育24 h后,ECV304细胞galectin-9基因和蛋白表达增加,且galectin-9表达与IFN-γ具有量效关系。PPARγ1基因转染抑制IFN-γ诱导galectin-9基因/蛋白表达,曲格列酮对上述作用无影响;PPARγ1基因转染和曲格列酮共刺激抑制IFN-γ诱导galectin-9基因/蛋白表达与单一PPARγ1基因转染效应相似。正常ECV304细胞PPARγ表达量低,而PPARγ基因过表达和活化不影响内源性PPARγ基因表达。结论:PPARγ1基因转染抑制IFN-γ诱导ECV304细胞galectin-9基因/蛋白表达可能是PPARγ基因发挥免疫调控作用的一个重要机制。AIM ： To investigate whether adenovirus - mediated mPPARγ1 gene overexpression inhibits IFN - γ -induced galectin-9 gene and protein expression in ECV304. METHODS： A replication- deficient recombinant adenovirus expression vector of mPPARγ1was constructed by using the AdEasy system. ECV304 were incubated for 24 h with 1 × 10^4 U/L, 5 × 10^4 U/L, 1 × 10^5 U/L and 2 × 10^5 U/L IFN - γ, respectively. ECV304 stimulated with 1 × 10^5 U/L IFN - γ were divided into 4 groups in random： P group （PPARγ1 gene overexpression）, T group （treated with troglitazone 40 μmol/L in DMSO）, PT group （ PPARγ1 gene overexpression ＋ troglitazone treatment） and control group. Changes of PPARγ and galectin - 9 in mRNA and protein levels in different groups and subgroups were investigated by RT - PCR and immunoblotting. RESULTS ： Galectin - 9 expression was very few in normal ECV304. IFN - γ induced the expression of galectin - 9 in ECV304. Degree of galectin - 9 expression increased with the dose of IFN - γ. PPARγ1 gene overexpression inhibited IFN - γ - induced galectin - 9 expression in ECV304. Galectin - 9 mRNA and protein expressions from PT group and P group were inhibited in similar degree （P 〉 0. 05 ）. However, this effect was not observed in troglitazone intervention （P 〉0. 05）. PPARγ expression was also very few in normal ECV304. PPARγ1 gene overexpression/activation had no effect on endogenous mPPARγ expression. CONCLUSION： This may partly contributed to the anti - inflammatory and immuno - regulatory effect of PPARγ1 gene overexpression by inhibiting IFN - γ - induced galectin - 9 gene and protein expression in ECV304.

关 键 词：过氧化物酶体增剂活化受体γ 动脉硬化 GALECTIN-9 

分 类 号：R363[医药卫生—病理学]