Characterisation of P2Y(12) receptor responsiveness to cysteinyl leukotrienes.
Characterisation of P2Y(12) receptor responsiveness to cysteinyl leukotrienes.
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DOI：
10.1371/journal.pone.0058305
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Woszczek G
中科院分区：
文献类型：
--
作者：
Foster HR;Fuerst E;Lee TH;Cousins DJ;Woszczek G
白三烯E4（LTE 4）是半胱氨酰白三烯（cysLT）中最稳定的一种，尽管在哮喘个体中它可能有效地诱导支气管收缩、气道高反应性和炎性细胞流入肺，但它与经典的1型和2型cysLT受体的结合较差。最近的一项研究表明，嘌呤能受体P2 Y12是哮喘小鼠模型中LTE 4介导的肺部炎症所必需的，并且是响应cysLT的信号。本研究的目的是验证人P2 Y12对半胱氨酰白三烯的反应性。使用在HEK 293、CHO细胞和人血小板中过表达的人CysLT 1、CysLT 2和P2 Y12的模型，并使用细胞内钙、cAMP和β-抑制蛋白募集测定来测量对不同激动剂的反应性。CysLT在过表达CysLT 1和CysLT 2的细胞中诱导浓度依赖性钙动员，但在表达P2 Y12或P2 Y12 + Gα16的细胞中未能诱导任何钙应答。相反，选择性P2 Y12激动剂ADP和2-MeS-ADP在表达P2 Y12 + Gα16的细胞中诱导特异性钙流。类似地，当分析细胞内cAMP和β-arrestin信号传导时，在表达P2 Y12的细胞中也观察到对2-MeS-ADP的特异性应答，但对cysLT没有。使用血小板作为表达P2 Y12的人原代细胞的模型，以分析通过P2 Y12受体或受体异二聚体的潜在信号传导和细胞活化，但未观察到特异性LTE 4应答。这些结果表明，LTE 4以及其他cysLT不激活通过P2 Y12起作用的细胞内信号传导，并表明另一种LTE 4特异性受体尚未被鉴定。
Leukotriene E4 (LTE4), the most stable of the cysteinyl leukotrienes (cysLTs), binds poorly to classical type 1 and 2 cysLT receptors although in asthmatic individuals it may potently induce bronchial constriction, airway hyperresponsiveness and inflammatory cell influx to the lung. A recent study has suggested that the purinergic receptor P2Y12 is required for LTE4 mediated pulmonary inflammation in a mouse model of asthma and signals in response to cysLTs. The aim of the study was to characterise the responsiveness of human P2Y12 to cysteinyl leukotrienes. Models of human CysLT1, CysLT2 and P2Y12 overexpressed in HEK293, CHO cells and human platelets were used and responsiveness to different agonists was measured using intracellular calcium, cAMP and β-arrestin recruitment assays. CysLTs induced concentration dependent calcium mobilisation in cells overexpressing CysLT1 and CysLT2 but failed to induce any calcium response in cells expressing P2Y12 or P2Y12+ Gα16. In contrast, selective P2Y12 agonists ADP and 2-MeS-ADP induced specific calcium flux in cells expressing P2Y12+ Gα16. Similarly, specific response to 2-MeS-ADP, but not to cysLTs was also observed in cells expressing P2Y12 when intracellular cAMP and β-arrestin signalling were analysed. Platelets were used as a model of human primary cells expressing P2Y12 to analyse potential signalling and cell activation through P2Y12 receptor or receptor heterodimers but no specific LTE4 responses were observed. These results show that LTE4 as well as other cysLTs do not activate intracellular signalling acting through P2Y12 and suggest that another LTE4 specific receptor has yet to be identified.
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DOI：
10.1164/ajrccm.155.4.9105062
发表时间：
1997-04-01
影响因子：
24.7
作者：
Diamant, Z;Hiltermann, JT;Sterk, PJ
通讯作者：
Sterk, PJ
影响因子：
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作者：
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影响因子：
4.8
作者：
OFFERMANNS, S;SIMON, MI
通讯作者：
SIMON, MI
影响因子：
3.6
作者：
Baurand, A;Eckly, A;Gachet, C
通讯作者：
Gachet, C
DOI：
10.1164/ajrccm.164.8.2102033
发表时间：
2001-10-15
影响因子：
24.7
作者：
Gauvreau, GM;Parameswaran, KN;O'Byrne, PM
通讯作者：
O'Byrne, PM