Mauban Joseph R H, Zacharia Joseph, Fairfax Seth, Wier Withrow Gil
Department of Physiology, School of Medicine, University of Maryland Baltimore, Baltimore, Maryland
Department of Physiology, School of Medicine, University of Maryland Baltimore, Baltimore, Maryland.
Am J Physiol Heart Circ Physiol. 2015 Jun 15;308(12):H1517-24. doi: 10.1152/ajpheart.00594.2014. Epub 2015 Apr 17.
Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Here, we used the PLC inhibitor, U-73122, and two other, specific inhibitors of PLC subtypes (PI-PLC and PC-PLC) to delineate the role of PLC in myogenic tone of pressurized murine mesenteric arteries. U-73122 inhibited depolarization-induced contractions (high external K(+) concentration), thus confirming reports of nonspecific actions of U-73122 and its limited utility for studies of myogenic tone. Edelfosine, a specific inhibitor of PI-PLC, did not affect depolarization-induced contractions but modulated myogenic tone. Because PI-PLC produces IP3, we investigated the effect of blocking IP3 receptor-mediated Ca(2+) release on myogenic tone. Incubation of arteries with xestospongin C did not affect tone, consistent with the virtual absence of Ca(2+) waves in arteries with myogenic tone. D-609, an inhibitor of PC-PLC and sphingomyelin synthase, strongly inhibited myogenic tone and had no effect on depolarization-induced contraction. D-609 appeared to act by lowering cytoplasmic Ca(2+) concentration to levels below those that activate contraction. Importantly, incubation of pressurized arteries with a membrane-permeable analog of DAG induced vasoconstriction. The results therefore mandate a reexamination of the signaling pathways activated by the Bayliss mechanism. Our results suggest that PI-PLC and IP3 are not required in maintaining myogenic tone, but DAG, produced by PC-PLC and/or SM synthase, is likely through multiple mechanisms to increase Ca(2+) entry and promote vasoconstriction.
肌源性张力是血管系统的一种内在特性,有助于血压控制和组织灌注。早期研究认为磷脂酶C(PLC)及其产物肌醇1,4,5 -三磷酸(IP3)和二酰甘油(DAG)在肌源性张力中起关键作用。在此,我们使用PLC抑制剂U - 73122以及另外两种PLC亚型的特异性抑制剂(PI - PLC和PC - PLC)来阐明PLC在加压小鼠肠系膜动脉肌源性张力中的作用。U - 73122抑制去极化诱导的收缩(高细胞外K⁺浓度),从而证实了关于U - 73122非特异性作用及其在肌源性张力研究中有限效用的报道。埃地福新,一种PI - PLC的特异性抑制剂,不影响去极化诱导的收缩,但调节肌源性张力。由于PI - PLC产生IP3,我们研究了阻断IP3受体介导的Ca²⁺释放对肌源性张力的影响。用西司他汀C孵育动脉不影响张力,这与具有肌源性张力的动脉中几乎不存在Ca²⁺波一致。D - 609,一种PC - PLC和鞘磷脂合酶的抑制剂,强烈抑制肌源性张力,对去极化诱导的收缩无影响。D - 609似乎通过将细胞质Ca²⁺浓度降低到激活收缩以下的水平起作用。重要的是,用DAG的膜渗透性类似物孵育加压动脉会诱导血管收缩。因此,这些结果要求重新审视由贝利斯机制激活的信号通路。我们的结果表明,维持肌源性张力不需要PI - PLC和IP3,但由PC - PLC和/或SM合酶产生的DAG可能通过多种机制增加Ca²⁺内流并促进血管收缩。