Heterozygous disruption of Flk-1 receptor leads to myocardial ischaemia reperfusion injury in mice: application of affymetrix gene chip analysis.
Heterozygous disruption of Flk-1 receptor leads to myocardial ischaemia reperfusion injury in mice: application of affymetrix gene chip analysis.
复制标题
Flk-1受体杂合破坏导致小鼠心肌缺血再灌注损伤：affymetrix基因芯片分析的应用。
DOI：
10.1111/j.1582-4934.2008.00269.x
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发表时间：
2008
影响因子：
5.3
通讯作者：
Maulik,N
中科院分区：
文献类型：
--
作者：
Thirunavukkarasu,M;Addya,S;Juhasz,B;Pant,R;Zhan,L;Surrey,S;Maulik,G;Menon,VP;Maulik,N
这项研究通过识别通过 Flk-1 受体触发缺血应激下心脏保护信号的血管内皮生长因子 (VEGF) 信号传导的潜在候选者，解决了一个重要的临床问题。将野生型 (WT) 和 Flk-1+/- 的离体工作小鼠心脏置于全身缺血 (I) 30 分钟。然后再灌注 2 小时 (R)。通过基线水平的定量实时 RT-PCR 检查，Flk-1+/- 心肌中的 Flk-1 mRNA 减少了近 50%。 2 小时后，Flk-1+/- 小鼠心脏在整个再灌注过程中左心室功能恢复有所减少（dp/dt 605 与 884）（P < 0.05）。再灌注 2 小时后，Flk-1+/- 中冠状动脉流量 (1.9 与 2.4 毫升) 和主动脉流量 (AF) (0.16 与 1.2 毫升) 减少。此外，在 Flk-1+/- 敲除 (KO) 小鼠中观察到梗死面积增加（38.4% 与 28.41%，P < 0.05）和心肌细胞凋亡（495 与 213）。我们还研究了缺血预处理（PC）（一种通过刺激 VEGF 信号通路来诱导心脏保护免受缺血再灌注损伤的新方法）是否可能在 Flk-1+/- 小鼠中发挥作用。我们发现，与 WT 相比，敲低 Flk-1 会导致 PC 的心脏保护作用显着降低。 Affymetrix 基因芯片分析表明，与 WT 小鼠相比，Flk-1+/- 小鼠在 IR 和预处理后缺血再灌注后重要基因下调。为了深入了解缺血性 PC 所涉及的潜在分子途径，我们使用 Ingenuity 途径分析工具确定了不同且重叠的生物过程。使用实时 RT-PCR 对选定的下调基因验证了支持 Affymetrix 结果的 mRNA 水平的独立证据，这些基因被认为在缺血性损伤后的心脏保护中发挥重要作用。总之，我们的数据首次表明，缺血性 PC 改变了杂合子 VEGFR-2/Flk-1 KO 小鼠的基因组反应，并消除了其对缺血心肌的心脏保护作用。
This study addresses an important clinical issue by identifying potential candidates of vascular endothelial growth factor (VEGF) signalling through the Flk‐1 receptor that trigger cardioprotective signals under ischaemic stress. Isolated working mouse hearts of both wild‐type (WT) and Flk‐1+/−were subjected to global ischaemia (I) for 30 min. followed by 2 hrs of reperfusion (R). Flk‐1+/−myocardium displayed almost 50% reduction in Flk‐1 mRNA as examined by quantitative real‐time RT‐PCR at the baseline level. Flk‐1+/−mouse hearts displayed reduction in left ventricular functional recovery throughout reperfusion (dp/dt 605versus884), after 2 hrs (P< 0.05). Coronary (1.9versus2.4 ml) and aortic flow (AF) (0.16versus1.2 ml) were reduced in Flk‐1+/−after 2 hrs of reperfusion. In addition, increased infarct size (38.4%versus28.41%,P< 0.05) and apoptotic cardiomyocytes (495versus213) were observed in Flk‐1+/−knockout (KO) mice. We also examined whether ischaemic preconditioning (PC), a novel method to induce cardioprotection against ischaemia reperfusion injury, through stimulating the VEGF signalling pathway might function in Flk‐1+/−mice. We found that knocking down Flk‐1 resulted in significant reduction in the cardioprotective effect by PC compared to WT. Affymetrix gene chip analysis demonstrated down‐regulation of important genes after IR and preconditioning followed by ischaemia reperfusion in Flk‐1+/−mice compared to WT. To get insight into the underlying molecular pathways involved in ischaemic PC, we determined the distinct and overlapping biological processes using Ingenuity pathway analysis tool. Independent evidence at the mRNA level supporting the Affymetrix results were validated using real‐time RT‐PCR for selected down‐regulated genes, which are thought to play important roles in cardioprotection after ischaemic insult. In summary, our data indicated for the first time that ischaemic PC modifies genomic responses in heterozygous VEGFR‐2/Flk‐1 KO mice and abolishes its cardioprotective effect on ischaemic myocardium.