高胰岛素状态下巨噬细胞PPARγ抗炎活性的变化  

Changes in macrophage PPAR gamma anti-inflammatory activities under high insulin levels

作　　者：张俊峰[1] 葛恒[1] 郭炳诗[2] 王长谦[1] 

机构地区：[1]上海交通大学医学院附属第二医院心内科,上海200001 [2]上海第二医科大学/中国科学院上海生命科学院健康科学中心,上海200025

出　　处：《心脏杂志》2006年第1期14-17,27,共5页Chinese Heart Journal

基　　金：国家自然基金项目资助(No.30070869)

摘　　要：目的探讨高胰岛素状态下巨噬细胞过氧化物酶体增殖物激活受体γ(PPARγ)抗炎活性的变化。方法体外培养THP1细胞,诱导分化为巨噬细胞,采用吡格列酮和不同浓度胰岛素分组干预,ELISA法测定细胞培养上清液中IL6、TNFα、MMP9浓度,GelatinZymography法测MMP9活性。半定量RTPCR、Westernblot检测巨噬细胞PPARγ基因、蛋白表达。结果PPARγ配体吡格列酮(20μmol/L)作用24h可显著降低巨噬细胞IL6(P<0.01)、TNFα、MMP9的分泌(P<0.05),抑制MMP9活性(P<0.05)。高胰岛素使吡格列酮抑制巨噬细胞分泌IL6、TNFα、MMP9的作用减弱,此作用与胰岛素浓度有关,呈浓度依赖性(0.25～0.5μmol/L)。而高胰岛素(0.1μmol/L)状态下巨噬细胞PPARγ基因、蛋白的表达均无显著变化。结论高胰岛素可减弱PPARγ配体抑制巨噬细胞分泌IL6、TNFα、MMP9的作用,但并未对巨噬细胞PPARγ基因、蛋白的表达产生影响,提示胰岛素对PPARγ的活性可能存在抑制。AIM To investigate the changes of the anti-inflammatory activities of macrophage PPAR gamma under high insulin levels. METHODS Human monocytic cell lines THP-1 were cultured and then induced into macrophage by PMA treatment. The differentiated monocytes were then cultured in medium of insulin at different concentrations treated with or without piogihazone. IL-6, TNF-α and MMP-9 levels in cell culture medium were measured by ELISA. MMP-9 activities were determined by gelatin zymography. Macrophage PPARγ mRNA and protein expression were detected by semi-quantitative RTPCR and Western blot, respectively. RESULTS Pioglitazone（20μmol/L） treatment for 24 hours significantly decreased the secretion of IL-6 （ P 〈 0.01 ） , TNF-α and MMP-9 （ P 〈 0.05 ） from macrophages as well as inhibited MMP-9 activity （P 〈 0.05 ）. High insulin（0.25μmol/L） partly counteracted the inhibitory action of pioglitazone on macrophage secretion of IL-6, TNF-α and MMP-9 in a concentration-dependent manner. High insulin （0.1 μmol/L）treatment had no significant effect on macrophage PPARγ mRNA or protein expression. CONCLUSION High insulin can counteract the inhibitory action of PPARγ agonists on macrophage secretion of IL-6, TNF-α and MMP-9, but has no significant effect on macrophage PPARγ mRNA or protein expression, suggesting that insulin may inhibit the activity of PPARγ.

关 键 词：胰岛素 巨噬细胞 过氧化物酶体增殖物激活爱体 炎症 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]