Analysis of inflammatory gene induction by oxidized phospholipids in vivo by quantitative real-time RT-PCR in comparison with effects of LPS
Analysis of inflammatory gene induction by oxidized phospholipids in vivo by quantitative real-time RT-PCR in comparison with effects of LPS
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DOI：
10.1016/s1537-1891(02)00172-6
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发表时间：
2002-04-01
影响因子：
4
通讯作者：
Leitinger, N
中科院分区：
文献类型：
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作者：
Kadl, A;Huber, J;Leitinger, N
氧化磷脂被认为在动脉粥样硬化和其他慢性炎症过程的发展中起作用。本研究采用实时定量逆转录聚合酶链式反应（RT-PCR）技术，分析了氧化l- α -棕榈酰-2-花生四烯酰基- asn -甘油-3-磷酸化胆碱（OxPAPC）诱导的炎症基因在体外和体内的表达情况。培养的人脐静脉内皮细胞(HUVEC)和monocyte-like U937细胞治疗与OxPAPC或lipopolysacchatide (LPS) 3 h。体内研究OxPAPC或有限合伙人通过静脉注入女性C57B1/6J老鼠和不同组织分离后3 h。我们发现OxPAPC和LPS诱导表达的早期生长反应因子1 (EGR-1)和单核细胞化学引诱物蛋白1 (MCP-1) HUVEC的我,鼠标MCP-1同系物,肝脏和心脏,有趣的是，OxPAPC而非LPS增加了血红素加氧酶1 （HO-1）在U937细胞、HUVEC、主动脉、心脏、肝脏和离体血细胞中的表达。LPS对E-selectin有选择性诱导作用，OxPAPC对E-selectin无选择性诱导作用。最后，用血小板活化因子（PAF）受体拮抗剂阻断oxpapc诱导的HO-1表达。我们得出结论，氧化磷脂在体内具有生物活性，并发挥特异性反应，诱导不同于LPS诱导的基因模式。此外，我们证明了实时定量RT-PCR技术是研究体内差异炎症基因诱导的合适工具。(C) 2002爱思唯尔科学有限公司版权所有。
Oxidized phospholipids are thought to play a role in the development of atherosclerosis and other chronic inflammatory processes. In this study, we analyzed the expression of inflammatory genes induced by oxidized L-alpha-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholin (OxPAPC) in vitro and in vivo using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Cultured human umbilical vein endothelial cells (HUVEC) and monocyte-like U937 cells were treated with OxPAPC or lipopolysacchatide (LPS) for 3 h. For in vivo studies, OxPAPC or LPS was injected intravenously into female C57B1/6J mice and different tissues were isolated after 3 h. We found that both OxPAPC and LPS induced expression of early growth response factor 1 (EGR-1) and monocyte chemoattractant protein 1 (MCP-1) in HUVEC and of JE, the mouse homologue of MCP-1, in liver and heart, Interestingly, OxPAPC but not LPS increased expression of heme oxygenase 1 (HO-1) in U937 cells, HUVEC, aorta, heart, liver, and isolated blood cells. In contrast, E-selectin was selectively induced by LPS, but not by OxPAPC. Finally, OxPAPC-induced expression of HO-1 was blocked by a platelet-activating factor (PAF) receptor antagonist. We conclude that oxidized phospholipids are biologically active in vivo and exert a specific response inducing a pattern of genes that is different from that induced by LPS. In addition, we demonstrate that the quantitative real-time RT-PCR technology is a proper tool to investigate differential inflammatory gene induction in vivo. (C) 2002 Elsevier Science Inc. All rights reserved.