PDF(Pubmed)
Abstract：
BACKGROUND: Peripheral blood oxygen monitoring via chemoreceptors in the carotid body (CB) is an integral function of the autonomic cardiorespiratory regulation. The presence of the purinergic P2Y12 receptor (P2Y12R) has been implicated in CB; however, the exact role of the receptor in O2 sensing and signal transduction is unknown.
METHODS: The presence of P2Y12R was established by immunoblotting, RT qPCR and immunohistochemistry. Primary glomus cells were used to assess P2Y12R function during hypoxia and hypercapnia, where monoamines were measured by HPLC; calcium signal was recorded utilizing OGB-1 and N-STORM Super-Resolution System. Ingravescent hypoxia model was tested in anaesthetized mice of mixed gender and cardiorespiratory parameters were recorded in control and receptor-deficient or drug-treated experimental animals.
RESULTS: Initially, the expression of P2Y12R in adult murine CB was confirmed. Hypoxia induced a P2Y12R-dependent release of monoamine transmitters from isolated CB cells. Receptor activation with the endogenous ligand ADP promoted release of neurotransmitters under normoxic conditions, while blockade disrupted the amplitude and duration of the intracellular calcium concentration. In anaesthetised mice, blockade of P2Y12R expressed in the CB abrogated the initiation of compensatory cardiorespiratory changes in hypoxic environment, while centrally inhibited receptors (i.e. microglial receptors) or receptor-deficiency induced by platelet depletion had limited influence on the physiological adjustment to hypoxia.
CONCLUSIONS: Peripheral P2Y12R inhibition interfere with the complex mechanisms of acute oxygen sensing by influencing the calcium signalling and the release of neurotransmitter molecules to evoke compensatory response to hypoxia. Prospectively, the irreversible blockade of glomic receptors by anti-platelet drugs targeting P2Y12Rs, propose a potential, formerly unrecognized side-effect to anti-platelet medications in patients with pulmonary morbidities.
METHODS: The presence of P2Y12R was established by immunoblotting, RT qPCR and immunohistochemistry. Primary glomus cells were used to assess P2Y12R function during hypoxia and hypercapnia, where monoamines were measured by HPLC; calcium signal was recorded utilizing OGB-1 and N-STORM Super-Resolution System. Ingravescent hypoxia model was tested in anaesthetized mice of mixed gender and cardiorespiratory parameters were recorded in control and receptor-deficient or drug-treated experimental animals.
RESULTS: Initially, the expression of P2Y12R in adult murine CB was confirmed. Hypoxia induced a P2Y12R-dependent release of monoamine transmitters from isolated CB cells. Receptor activation with the endogenous ligand ADP promoted release of neurotransmitters under normoxic conditions, while blockade disrupted the amplitude and duration of the intracellular calcium concentration. In anaesthetised mice, blockade of P2Y12R expressed in the CB abrogated the initiation of compensatory cardiorespiratory changes in hypoxic environment, while centrally inhibited receptors (i.e. microglial receptors) or receptor-deficiency induced by platelet depletion had limited influence on the physiological adjustment to hypoxia.
CONCLUSIONS: Peripheral P2Y12R inhibition interfere with the complex mechanisms of acute oxygen sensing by influencing the calcium signalling and the release of neurotransmitter molecules to evoke compensatory response to hypoxia. Prospectively, the irreversible blockade of glomic receptors by anti-platelet drugs targeting P2Y12Rs, propose a potential, formerly unrecognized side-effect to anti-platelet medications in patients with pulmonary morbidities.
摘要：
背景：通过颈动脉体（CB）中的化学感受器监测外周血氧是自主性心肺调节的不可或缺的功能。嘌呤能P2Y12受体（P2Y12R）的存在与CB有关；然而，受体在O2传感和信号转导中的确切作用尚不清楚。
方法：通过免疫印迹确定P2Y12R的存在，RT一qPCR和免疫组织化学。原代血管球细胞用于评估P2Y12R在缺氧和高碳酸血症期间的功能，其中通过HPLC测量单胺;使用OGB-1和N-STORM超分辨率系统记录钙信号。在性别混合的麻醉小鼠中测试了严重缺氧模型，并在对照和受体缺陷或药物治疗的实验动物中记录了心肺参数。
结果：最初，证实P2Y12R在成年鼠CB中的表达。缺氧诱导P2Y12R依赖性的单胺递质从分离的CB细胞释放。在常氧条件下，内源性配体ADP的受体激活促进了神经递质的释放，而阻断破坏了细胞内钙浓度的幅度和持续时间。在麻醉小鼠中，在CB中表达的P2Y12R的阻断消除了低氧环境中代偿性心肺变化的开始，而中枢抑制受体（即小胶质细胞受体）或血小板耗竭诱导的受体缺乏对缺氧的生理调节影响有限。
结论：外周P2Y12R抑制通过影响钙信号和神经递质分子的释放来干扰急性氧敏感的复杂机制，从而引起对缺氧的代偿反应。Prospective,靶向P2Y12Rs的抗血小板药物对glomic受体的不可逆阻断，提出一个潜力，以前未发现的抗血小板药物对肺部疾病患者的副作用。
方法：通过免疫印迹确定P2Y12R的存在，RT一qPCR和免疫组织化学。原代血管球细胞用于评估P2Y12R在缺氧和高碳酸血症期间的功能，其中通过HPLC测量单胺;使用OGB-1和N-STORM超分辨率系统记录钙信号。在性别混合的麻醉小鼠中测试了严重缺氧模型，并在对照和受体缺陷或药物治疗的实验动物中记录了心肺参数。
结果：最初，证实P2Y12R在成年鼠CB中的表达。缺氧诱导P2Y12R依赖性的单胺递质从分离的CB细胞释放。在常氧条件下，内源性配体ADP的受体激活促进了神经递质的释放，而阻断破坏了细胞内钙浓度的幅度和持续时间。在麻醉小鼠中，在CB中表达的P2Y12R的阻断消除了低氧环境中代偿性心肺变化的开始，而中枢抑制受体（即小胶质细胞受体）或血小板耗竭诱导的受体缺乏对缺氧的生理调节影响有限。
结论：外周P2Y12R抑制通过影响钙信号和神经递质分子的释放来干扰急性氧敏感的复杂机制，从而引起对缺氧的代偿反应。Prospective,靶向P2Y12Rs的抗血小板药物对glomic受体的不可逆阻断，提出一个潜力，以前未发现的抗血小板药物对肺部疾病患者的副作用。
