A novel in vitro ischemia/reperfusion injury model
A novel in vitro ischemia/reperfusion injury model
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DOI：
10.1007/s12272-009-1316-9
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发表时间：
2009-03-01
影响因子：
6.7
通讯作者：
Lee, Yong Woo
中科院分区：
文献类型：
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作者：
Lee, Won Hee;Kang, Sungkwon;Lee, Yong Woo
血流再灌注发生在许多情况下，如中风和器官移植，极大地增加了组织损伤，造成了比长期缺血更严重的损害。在本研究中，我们设计了一种新型的双层平行板流动室(PPFC)来建立体外缺血/再灌注(I/R)损伤模型，并观察了I/R对人微血管内皮细胞(HMEC-1)炎症反应的影响。采用实时定量RT-PCR方法检测HMEC-1细胞中促炎介质白介素6(IL-6)、单核细胞趋化蛋白-1(MCP-1)、细胞间黏附分子-1(ICAM-1)、E-选择素和血管细胞黏附分子-1(VCAM-1)的表达。细胞还被抗氧化剂吡咯烷二硫代氨基甲酸酯(PDTC)预处理，以验证体外I/R损伤中的氧化机制。观察生理流对HMCE-1细胞形态和促炎介质表达的影响。相反，I/R显著上调HMEC-1中促炎介质的表达。此外，PDTC的预处理显著减少了I/R介导的促炎介质的过度表达。本研究的数据表明，我们新设计的PPFC可以作为一种有效的体外细胞培养模型系统来开发针对缺血/再灌注(I/R)损伤的新药。
The reperfusion of blood flow occurred in a number of conditions such as stroke and organ transplantation immensely augments tissue injury and causes more severe damage than prolonged ischemia. In the present study, we designed a novel double-layer parallel-plate flow chamber (PPFC) to develop an in vitro ischemia/reperfusion (I/R) injury model and examined the effects of I/R on inflammatory responses in human microvascular endothelial cells (HMEC-1). The expression of pro-inflammatory mediators, such as interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), E-selectin, and vascular cell adhesion molecule-1 (VCAM-1) in HMEC-1 was measured by quantitative real-time RT-PCR. The cells were also pre-treated with antioxidant pyrrolidine dithiocarbamate (PDTC) to verify involvement of an oxidative mechanism in I/R injury in vitro. The morphological changes and attenuated expression of pro-inflammatory mediators were observed in HMCE-1 exposed to the physiological flow. In contrast, I/R markedly and significantly up-regulated expression of pro-inflammatory mediators in HMEC-1. Additionally, pretreatment with PDTC significantly reduced I/R-mediated overexpression of pro-inflammatory mediators. The data from the present study provide evidence demonstrating that our newly designed PPFC can be utilized as an effective in vitro cell culture model system to develop new drugs specifically targeting against ischemia/reperfusion (I/R) injury.