NTPDase1 (CD39) controls nucleotide-dependent vasoconstriction in mouse
NTPDase1 (CD39) controls nucleotide-dependent vasoconstriction in mouse
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DOI：
10.1093/cvr/cvp265
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发表时间：
2010-01-01
影响因子：
10.8
通讯作者：
Sevigny, Jean
中科院分区：
文献类型：
--
作者：
Kauffenstein, Gilles;Drouin, Annick;Sevigny, Jean
目的 细胞外核苷酸是血管活性分子。这些分子的浓度由核酸外切酶调节。在本研究中，我们利用 Entpd1(-/-) 小鼠研究了血管核酸外切酶 NTPDase1 在核苷酸的血管收缩作用中的作用。 方法和结果 使用免疫荧光、酶组织化学和 HPLC 分析来评估动脉和分离的血管平滑肌细胞 (VSMC) 中的 NTPDase 表达和活性。在体外评估血管反应性，并在静脉注射核苷酸后记录麻醉小鼠的平均动脉血压。输液。通过RT-PCR测定VSMC中核苷酸受体的表达。与对照小鼠相比，Entpd1(-/-) 小鼠在原位血管壁和 VSMC 中显示出明显的核苷酸酶活性缺陷。在 Entpd1(-/-) 小鼠的主动脉环中，UDP 和 UTP 诱导了有效且持久的收缩，与野生型环中获得的微弱反应形成鲜明对比。这种收缩通过 P2Y(6) 受体的激活发生，并且独立于其他尿嘧啶核苷酸反应受体（P2Y(2) 和 P2Y(4)）。 UDP 体内输注会增加血压，并且这种效应在 Entpd1(-/-) 小鼠中得到加强。此外，Entpd1(-/-)小鼠的受压肠系膜动脉表现出增强的肌源性反应，这与内源性释放的核苷酸的局部浓度较高相一致。这种作用可被P2受体拮抗剂RB-2抑制。结论 NTPDase1是调节VSMC表面核苷酸代谢的主要酶，因此有助于核苷酸对血管张力的局部调节。
Aims Extracellular nucleotides are vasoactive molecules. The concentrations of these molecules are regulated by ectonucleotidases. In this study, we investigated the role of the blood vessel ectonucleotidase NTPDase1, in the vasoconstrictor effect of nucleotides using Entpd1(-/-) mice.Methods and results Immunofluorescence, enzyme histochemistry, and HPLC analysis were used to evaluate both NTPDase expression and activity in arteries and isolated vascular smooth muscle cells (VSMCs). Vascular reactivity was evaluated in vitro and mean arterial blood pressure was recorded in anesthetized mice after nucleotide i.v. infusion. Expression of nucleotide receptors in VSMCs was determined by RT-PCR. Entpd1(-/-) mice displayed a dramatic deficit of nucleotidase activity in blood vessel wall in situ and in VSMCs in comparison to control mice. In aortic rings from Entpd1(-/-) mice, UDP and UTP induced a potent and long-lasting constriction contrasting with the weak response obtained in wild-type rings. This constriction occurred through activation of P2Y(6) receptor and was independent of other uracil nucleotide-responding receptors (P2Y(2) and P2Y(4)). UDP infusion in vivo increased blood pressure and this effect was potentiated in Entpd1(-/-) mice. In addition, pressurized mesenteric arteries from Entpd1(-/-) mice displayed an enhanced myogenic response, consistent with higher local concentrations of endogenously released nucleotides. This effect was inhibited by the P2 receptor antagonist RB-2.Conclusion NTPDase1 is the major enzyme regulating nucleotide metabolism at the surface of VSMCs and thus contributes to the local regulation of vascular tone by nucleotides.