Sullivan Jennifer C, Giulumian Ararat D, Pollock David M, Fuchs Leslie C, Pollock Jennifer S
Vascular Biology Center, Medical College of Georgia, 1459 Laney-Walker Boulevard, Augusta, GA 30912, USA.
Am J Physiol Heart Circ Physiol. 2002 Aug;283(2):H658-63. doi: 10.1152/ajpheart.00073.2002.
Previously we have demonstrated functional nitric oxide synthase (NOS) 1 in large arteries. Because resistance arteries largely determine blood pressure, this study examined whether functional NOS 1 also exists in resistance arteries. Phenylephrine (PE) contraction was measured in the absence and presence of the NOS 1 inhibitor N(5)-(1-imino-3-butenyl)-L-ornithine (VNIO) in isolated mesenteric resistance arteries (endothelium intact and denuded) from Sprague-Dawley rats. For NOS 1 activity and expression, the mesenteric arterial bed was separated into cytosolic and particulate fractions. NOS activity was assayed by measuring the conversion of [(3)H]arginine to [(3)H]citrulline inhibited by a nonselective NOS inhibitor or VNIO. VNIO increased PE sensitivity in endothelium-intact and -denuded arteries. In cytosolic and particulate fractions of the arterial bed, approximately 40% of NOS activity was inhibited by VNIO. Immunoprecipitation and Western blot analysis revealed two NOS 1 immunoreactive bands. One band corresponded to the rat brain isoform, whereas the second was of a slightly lower molecular mass. The cytosolic fraction contained both isoforms; however, the particulate fraction had only the lower molecular mass form. These studies demonstrate the existence of functional NOS 1 in resistance arteries.
此前我们已在大动脉中证实了功能性一氧化氮合酶(NOS)1的存在。由于阻力动脉在很大程度上决定血压,本研究检测了阻力动脉中是否也存在功能性NOS 1。在来自Sprague-Dawley大鼠的离体肠系膜阻力动脉(内皮完整和去内皮)中,在不存在和存在NOS 1抑制剂N(5)-(1-亚氨基-3-丁烯基)-L-鸟氨酸(VNIO)的情况下测量去氧肾上腺素(PE)收缩。对于NOS 1活性和表达,将肠系膜动脉床分离为胞质和微粒体部分。通过测量[(3)H]精氨酸向[(3)H]瓜氨酸的转化来测定NOS活性,该转化受非选择性NOS抑制剂或VNIO抑制。VNIO增加了内皮完整和去内皮动脉中PE的敏感性。在动脉床的胞质和微粒体部分,约40%的NOS活性被VNIO抑制。免疫沉淀和蛋白质印迹分析显示两条NOS 1免疫反应带。一条带对应大鼠脑同工型,而另一条带的分子量略低。胞质部分含有两种同工型;然而,微粒体部分仅具有较低分子量形式。这些研究证明了阻力动脉中存在功能性NOS 1。