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Effect of human 15-lipoxygenase-l metabolites on vascular function in mouse mesenteric arteries and hearts

机译:人15-脂氧基酶-L代谢物对小鼠肠系膜动脉和心脏血管功能的影响

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摘要

Lipoxygenases regulate vascular function by metabolizing arachidonic acid (AA) to dilator eicosanoids. Previously, we showed that endothelium-targeted adenoviral vector-mediated gene transfer of the human 15-lipoxygenase-l (h15-LO-l) enhances arterial relaxation through the production of vasodila-tory hydroxyepoxyeicosatrienoic acid (HEETA) and trihydroxyeicosatrienoic acid (THETA) metabolites. To further define this function, a transgenic (Tg) mouse line that overexpresses h1 5-LO-l was studied. Western blot, immunohistochemistry and RT-PCR results confirmed expression of 15-LO-l transgene in tissues, especially high quantity in coronary arterial wall, of Tg mice. Reverse-phase HPLC analysis of [14C]-AA metabolites in heart tissues revealed enhanced 15-HETE synthesis in Tg vs. WT mice. Among the 15-LO-l metabolites, 15-HETE, erythro-13-H-14,15-EETA, and 11(R),12(S),15(S)-THETA relaxed the mouse mesenteric arteries to the greatest extent. The presence of h15-LO-l increased acetylcholine-and AA-mediated relaxation in mesenteric arteries of Tg mice compared to WT mice. 15-LO-l was most abundant in the heart; therefore, we used the Langendorff heart model to test the hypothesis that elevated 15-LO-l levels would increase coronary flow following a short ischemia episode. Both peak flow and excess flow of reperfused hearts were significantly elevated in hearts from Tg compared to WT mice being 2.03 and 3.22 times greater, respectively. These results indicate that h15-LO-l -derived metabolites are highly vasoactive and may play a critical role in regulating coronary blood flow.
机译:脂氧基酶通过将花生酸(AA)代谢到扩张器籽糖烷来调节血管功能。以前,我们表明,通过生产血管发水机 - Tory羟基氧基己酸(Heeta)和三羟基己二烯酸(Heeta)和三羟基己二烯酸(Heeta)和三羟基己二烯酸(Theta)增强动脉弛豫的内皮靶向腺嘌呤载体介导的基因转移增强动脉弛豫代谢物。为了进一步定义该功能,研究了过表达H1 5-LO-L的转基因(TG)小鼠线。 Western印迹,免疫组化和RT-PCR结果证实了TG小鼠组织中15-LO-L转基因的表达,尤其是冠状动脉壁,尤其是TG小鼠的表达。 [14C] -AA代谢物的反相HPLC分析在心脏组织中显示出增强的15-HETE合成在TG与WT小鼠中。在15-LO-L代谢物中,15-HETE,eryTHRO-13-H-14,15- eETA和11(R),12(S),15(S)-THETA放松了最大的小鼠肠系膜动脉程度。与WT小鼠相比,H15-LO-L的存在增加了Tg小鼠的肠系膜中的乙酰胆碱 - 和AA介导的弛豫。 15-lo-l在内心中最丰富;因此,我们使用Langendorff心脏模型来测试升高的15-LO-L水平的假设将增加冠状动脉术后的冠状动脉流量。与WT小鼠相比,Tg的再灌注心脏的峰流量和再灌注心脏的过量流量分别为2.03和3.22倍。这些结果表明,H15-LO-L长长的代谢物高度血管活性,并且可能在调节冠状动脉血流方面发挥关键作用。

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