Department of Physiology, School of Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, Australia;
Am J Physiol Regul Integr Comp Physiol. 2013 Oct 15;305(8):R917-26. doi: 10.1152/ajpregu.00225.2013. Epub 2013 Aug 15.
In pregnancy, α-adrenoceptor-mediated vasoconstriction is augmented in uterine radial arteries and is accompanied by underlying changes in smooth muscle (SM) Ca(2+) activity. This study aims to determine the Ca(2+) entry channels associated with altered vasoconstriction in pregnancy, with the hypothesis that augmented vasoconstriction involves transient receptor potential canonical type-3 (TRPC3) and L- and T-type voltage-dependent Ca(2+) channels. Immunohistochemistry showed TRPC3, L-type Cav1.2 (as the α1C subunit), T-type Cav3.1 (α1G), and Cav3.2 (α1H) localization to the uterine radial artery SM. Fluorescence intensity of TRPC3, Cav1.2, and Cav3.2 was increased, and Cav3.1 decreased in radial artery SM from pregnant rats. Western blot analysis confirmed increased TRPC3 protein expression in the radial artery from pregnant rats. Pressure myography incorporating pharmacological intervention to examine the role of these channels in uterine radial arteries showed an attenuation of phenylephrine (PE)-induced constriction with Pyr3 {1-[4-[(2,3,3-trichloro-1-oxo-2-propen-1-yl)amino]phenyl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid}-mediated TRPC3 inhibition or with nifedipine-mediated L-type channel block alone in vessels from pregnant rats; both effects of which were diminished in radial arteries from nonpregnant rats. Combined TRPC3 and L-type inhibition attenuated PE-induced constriction in radial arteries, and the residual vasoconstriction was reduced and abolished with T-type channel block with NNC 55-0396 in arteries from nonpregnant and pregnant rats, respectively. With SM Ca(2+) stores depleted and in the presence of PE, nifedipine, and NNC 55-0396, blockade of TRPC3 reversed PE-induced constriction. These data suggest that TRPC3 channels act synergistically with L- and T-type channels to modulate radial artery vasoconstriction, with the mechanism being augmented in pregnancy.
在妊娠期间,子宫放射状动脉中的 α-肾上腺素能受体介导的血管收缩增强,并伴有平滑肌(SM)Ca(2+)活性的潜在变化。本研究旨在确定与妊娠中血管收缩改变相关的 Ca(2+)内流通道,假设增强的血管收缩涉及瞬时受体电位经典型 3(TRPC3)和 L 型和 T 型电压依赖性 Ca(2+)通道。免疫组织化学显示 TRPC3、L 型 Cav1.2(作为 α1C 亚基)、T 型 Cav3.1(α1G)和 Cav3.2(α1H)定位于子宫放射状动脉 SM。从妊娠大鼠的放射状动脉中,TRPC3、Cav1.2 和 Cav3.2 的荧光强度增加,而 Cav3.1 则减少。Western blot 分析证实了妊娠大鼠放射状动脉中 TRPC3 蛋白表达的增加。结合药理学干预的压力血管造影术检查这些通道在子宫放射状动脉中的作用表明,用 Pyr3(1-[4-[(2,3,3-三氯-1-氧代-2-丙烯基)氨基]苯基]-5-(三氟甲基)-1H-吡唑-4-羧酸)抑制 TRPC3 或单独用硝苯地平抑制 L 型通道可减弱苯肾上腺素(PE)诱导的收缩;这两种作用在非妊娠大鼠的放射状动脉中均减弱。联合 TRPC3 和 L 型抑制减弱了 PE 诱导的放射状动脉收缩,而在非妊娠和妊娠大鼠的动脉中,用 NNC 55-0396 阻断 T 型通道可进一步减少和消除剩余的血管收缩。在 SM Ca(2+)储存耗尽的情况下,在存在 PE、硝苯地平和 NNC 55-0396 的情况下,TRPC3 阻断逆转了 PE 诱导的收缩。这些数据表明,TRPC3 通道与 L 型和 T 型通道协同作用,调节放射状动脉血管收缩,其机制在妊娠中增强。