15-Hydroxy-5,8,11,13-eicosatetraenoic acid inhibits human vascular cyclooxygenase. Potential role in diabetic vascular disease.
15-Hydroxy-5,8,11,13-eicosatetraenoic acid inhibits human vascular cyclooxygenase. Potential role in diabetic vascular disease.
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15-Hydroxy-5,8,11,13-二十碳四烯酸抑制人血管环氧合酶。
DOI：
10.1172/jci112277
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Stuart,MJ
中科院分区：
文献类型：
--
作者：
Setty,BN;Stuart,MJ
人类脐动脉将花生四烯酸转化为三种羟基二十碳四烯酸（HETE）以及三尖杉酯碱。通过反相高压液相色谱法和气相色谱-质谱法鉴定了单HETE为15-HETE和11-HETE。15-动脉段中的HETE似乎主要通过15-脂氧合酶途径产生，而11-HETE和推测的二-HETE是环氧合酶的产物。去甲二氢愈创木酸是一种脂氧合酶抑制剂，刺激前列腺素类的产生，同时抑制15-HETE的形成。这些结果表明，15-HETE可能作为前列环素的内源性调节剂。在人脐动脉微粒体中，发现15-HETE以浓度依赖性方式抑制6-酮-前列腺素F1 α和总前列腺素类的产生（中位抑制常数[IC 50]分别为52 +/- 3和63 +/- 4 μ M）。然而，野牡丹素的相对分布不受影响，表明作用部位是环氧合酶。动力学分析表明，15-HETE是酶的竞争性抑制剂。虽然最大速度没有发生变化，但表观Km显著不同（对照组为9.3 +/- 6.9 μ M [1 SD]，而15-HETE处理的酶为37.6 +/- 17.7 μ M）。此外，使用培养的牛内皮细胞证实了15-HETE对前列环素产生的抑制作用。在该细胞系统中，15-HETE不仅抑制内源性前列环素的产生，而且抑制外源性[1- 14 C]花生四烯酸向前列环素的转化（IC 50为40 +/- 17 μ M）。对花生四烯酸释放无影响。为了研究我们在体外发现15-HETE抑制前列环素的产生是否与体内情况相关，我们对从糖尿病环境中获得的血管进行了最终研究。我们发现，与对照组新生儿（0.77 +/- 0.22; P <0.01）相比，糖尿病母亲的婴儿血管中15-HETE的产生显著增加（1.14 +/- 0.26 pmol/mg）。同时观察到前列环素产生减少（糖尿病母亲的婴儿为51.6 +/- 12.6 pmol/mg，对照组为71 +/- 22.3 pmol/mg）。此外，还注意到这两种类二十烷酸之间的负相关。我们的研究结果表明15-HETE对前列环素的产生具有潜在的体内调节作用。
Human umbilical arteries converted arachidonic acid to three hydroxyeicosatetraenoic acids (HETEs) as well as prostaglandins. The mono-HETEs have been identified by reverse-phase high pressure liquid chromatography and gas chromatography-mass spectroscopy as 15-HETE and 11-HETE. 15-HETE in arterial segments appears to be derived mainly via the 15-lipoxygenase pathway, whereas 11-HETE, and the presumed di-HETE(s) were products of cyclooxygenase. Nordihydroguaiaretic acid, a lipoxygenase inhibitor, stimulated prostanoid production with a concomitant inhibition of 15-HETE formation. These results suggested that 15-HETE may function as an endogenous regulator of prostacyclin. In human umbilical arterial microsomes, 15-HETE was found to inhibit 6-keto-prostaglandin F1 alpha and total prostanoid production in a concentration-dependent manner (median inhibition constant [IC50] of 52 +/- 3 and 63 +/- 4 microM respectively). The relative distribution of prostaglandins, however, remained unaffected, indicating that the site of action was cyclooxygenase. Kinetic analysis revealed that 15-HETE was a competitive inhibitor of the enzyme. Although no changes in maximum velocity occurred, the apparent Km was significantly different (9.3 +/- 6.9 microM [1 SD] for control vs. 37.6 +/- 17.7 microM for the 15-HETE-treated enzyme). Furthermore, the inhibitory effect of 15-HETE on prostacyclin production was confirmed using cultured bovine endothelial cells. In this cell system, not only did 15-HETE inhibit endogenous prostacyclin production, but also the conversion of exogenous [1-14C]arachidonic acid to prostacyclin (IC50 of 40 +/- 17 microM). No effect on arachidonic acid release was noted. To investigate whether our in vitro finding that 15-HETE inhibited prostacyclin production could be relevant to the in vivo situation, our final studies were performed on vasculature obtained from the diabetic milieu. We found that the production of 15-HETE was significantly increased in vasculature obtained from the infant of the diabetic mother (1.14 +/- 0.26 pmol/mg) when compared to control neonates (0.77 +/- 0.22; P less than 0.01). A concomitant decrease in prostacyclin production was seen (51.6 +/- 12.6 pmol/mg in infants of diabetic mothers vs. 71 +/- 22.3 in controls). Moreover, an inverse correlation between these two eicosanoids was also noted. Our results suggest a potential in vivo regulatory role for 15-HETE on prostacyclin production.Images