摘要
目的探讨过氧化物酶增殖物激活受体-γ(PPAR-γ)在调节急性冠状动脉综合征(ACS)患者外周血单核细胞源性巨噬细胞(MDM)表达基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶抑制物-1(TIMP-1)中的作用及可能机制。方法选取ACS患者48例(ACS组)、对照者28例(对照组),提取外周血单个核细胞,刺激培养为MDM。随机将ACS组及对照组MDM分别分为罗格列酮0、1、10和20umol/L亚组,分别于相应亚组加入血清、1、10和20umol/L浓度的罗格列酮干预。然后测定各亚组MDM上清液中MMP-9、TIMP-1浓度及MDM细胞中PPAR-γ MMP-9、TIMP-1mRNA表达强度,并用免疫组织化学法测定核因子-kB(NF-kB)蛋白表达变化。分析比较ACS组与对照组间不同浓度罗格列酮干预亚组表达PPAR-γ、MMP-9、TIMP-1 mRNA及MMP-9、TIMP-1、核因子-k B P65(NF—k B P65)蛋白水平上的差异。结果罗格列酮干预后,ACS组及对照组MDM中PPAR-1mRNA表达明显上调[PPAR-1mRNA,ACS组:0.25±0.05,0.26±0.07,0.74±0.12,0.86±0.12,1.16±0.26,1、10、20umo]/L亚组与干预前及0umol/L亚组比JP2较P均〈0.01;对照组:0.39±0.06,0.37±0.04,0.99±0.10,0.99±0.09,1.00±0.12,1、10、20umol/L亚组与干预前及0umol/L亚组比较P均〈0.01;数据排列顺序为干预前、0、1、10、20umol/L亚组,下同],在ACS组中表达强度与罗格列酮浓度呈正比关系。ACS组及对照组MDM上清液中MMP-9浓度下降[MMP-9浓度(mmol/L),ACS组:231±51,215±47,181±36,172±30,151±24,1、10、20umol/L亚组与干预前及0umol/L亚组比较P均〈0.01;对照组:138±15,127±15,119±13,112±17,107±18,1umol/L亚组与干预前比较P〈0.05,10、20umol/L亚组与干预前比较P均〈0.01,10umol/L亚组与0umol/L亚组比较P〈0.05,20umol/L亚组与0umol/L亚组比较P〈0.01];MDM中MMP-9mRNA表达下调[MMP-9mRNA,ACS组:0.67±0.11,0.62±0.10,0.54±0.05,0.48±0.10,0.41±0.06,1、10、20umol/L亚组与干预前比较P均〈0.01,10、20umol/L亚组与0umol/L亚组比较P均〈0.01,1umoL/L亚组与0umol/L亚组比较P〈0.05;对照组:0.24±0.08,0.26±0.09,0.20±0.05,0.22±0.05,0.20±0.04,10、20umol/L亚组与干预前及0umol/L亚组比较P均〈0.01,1umol/L亚组与干预前比较P〈0.05],ACS组中下调程度与罗格列酮浓度呈反比关系。ACS组和对照组上清液中TIMP-1浓度及MDM中TIMP-1 mRNA的表达无明显变化。ACS组和对照组MDM中NF—kB P65表达下调[NF—kB P65 OD值,ACS组:0.42±0.09,0.41±0.09,0.34±0.04,0.34±0.04,0.32±0.05,1、10、20umol/L亚组与干预前及0umol/L亚组比较P均〈0.01;对照组:0.28±0.03,0.25±0.04,0.23±0.03,0.23±0.04,0.22±0.05,1、10、20umol/L亚组与干预前及0umol/L亚组比较P均〈0.05],不同浓度罗格列酮亚组间差异无统计学意义。结论罗格列酮可能通过上调PPAR-1表达,减低ACS组NF—kB水平,而下调ACS患者MDM表达MMP-9,但TMPI-1表达不受此调节。罗格列酮可能具有稳定ACS患者动脉粥样斑块的作用。
Objective Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS) . Metalloprotease (MMPs) secreted by monocyte/ macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-γ (PPAR-γ) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-γ in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS. Methods Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony- stimulating factor (0. 1 ug/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 umol/L) for 48 hours. The concentrations of MMP-9 and TIMP-I in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-γ, MMP-9 by RT-PCR and nuclear factor-KB P65 (NF-KB P65 ) expression by immunohistochemistry. Results PPAR-3, mRNA expression was significantly lower while NF-KB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group ( all P 〈 0. 05 ). After rosiglitazone intervention, PPAR-γ mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulatio, of NF-KB P65 expression in both groups. Conclusion Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-γ expression, and by downregulaiting NF-KB expression in MDMs isolated from patients with ACS.
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2009年第8期739-745,共7页
Chinese Journal of Cardiology
基金
深圳市科技基金项目(JH200507120929A)
龙岗区科技基金资助(200514)