A flow cytometric analysis of the inhibition of platelet reactivity due to nitrite reduction by deoxygenated erythrocytes.
A flow cytometric analysis of the inhibition of platelet reactivity due to nitrite reduction by deoxygenated erythrocytes.
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DOI：
10.1371/journal.pone.0092435
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Schechter AN
中科院分区：
文献类型：
--
作者：
Akrawinthawong K;Park JW;Piknova B;Sibmooh N;Fucharoen S;Schechter AN
一氧化氮(NO)是一种小分子气体，长期以来一直被认为是一种有效的血小板功能抑制物，但NO抑制血小板功能的生理和病理意义尚未很好地阐明。我们最近发现，在红细胞存在的情况下，向富含血小板的血浆中加入亚硝酸盐可以抑制血小板聚集，通过P-选择素的表达和cGMP的产生来衡量，亚硝酸盐+红细胞的这种抑制作用因红细胞的脱氧而增强。为了研究不同氧水平下亚硝酸盐对血小板的影响，我们用流式细胞仪检测了血小板膜表面标志物的活化情况。在不同的血细胞比容和血氧水平下，检测ADP、胶原和凝血酶刺激对血小板膜P-选择素和活化的GPIIb/IIIa表达的影响。当血细胞比容大于23%，血氧水平低于49μ时，亚硝酸盐(0.1mMm~1.0mmHgM)可显著降低血小板膜表面标志的百分比。亚硝酸盐的抑制作用随着红细胞压积的升高和血氧饱和度的降低而增强。NO清除剂C-PTIO可拮抗亚硝酸盐+红细胞对血小板的抑制作用，而NO合成酶和黄嘌呤氧化还原酶抑制剂则无此作用。这些结果支持这样一种观点，即循环中的亚硝酸盐通过还原成NO来降低部分脱氧红细胞存在时的血小板反应性，这也可能解释了动脉和静脉血栓形成之间的某些差异，并直接支持了脱氧血红蛋白在这一过程中的作用。我们相信，我们的流式细胞仪分析为识别参与这些效应的单个分子过程提供了可能性。
Nitric oxide (NO), a small gas molecule, has long been known to be a potent inhibitor of platelet function but the physiological and pathological implications of platelet inhibition by NO have not been well clarified. We recently showed that the addition of nitrite to platelet-rich plasma in the presence of erythrocytes could inhibit platelet aggregation and this inhibitory effect of nitrite + erythrocytes was enhanced by deoxygenation of erythrocytes as measured by P-selectin expression and cGMP production. In order to study the nitrite effect on platelets at different oxygen levels, we used the flow cytometric assays to detect platelet membrane surface markers upon activation. The P-selectin and activated gpIIb/IIIa expression on platelet membranes in response to ADP, collagen and thrombin stimulation was measured at various hematocrit and oxygen levels. Nitrite (0.1 to 1.0 μM) significantly decreased the percentage of these surface markers on the platelet membrane at the hematocrit values above 23% and oxygen levels lower than 49 mmHg. The inhibitory effect of nitrite was augmented by increasing hematocrit values and decreasing oxygen saturation. C-PTIO (an NO scavenger) prevented the platelet inhibition by nitrite + erythrocytes whereas the inhibitors of NO synthase and xanthine oxidoreductase had no effect. These results support the proposal that circulating nitrite decreases platelet reactivity in the presence of partially deoxygenated erythrocytes through its reduction to NO, which may also explain certain differences between arterial and venous thrombosis and support directly the role of deoxyhemoglobin in this process. We believe that our flow cytometric assays offer a possibility to identify the individual molecular process involved in these effects.
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亚硝酸盐抑制血小板取决于红细胞和脱氧。
影响因子：
3.7
作者：
Srihirun S;Sriwantana T;Unchern S;Kittikool D;Noulsri E;Pattanapanyasat K;Fucharoen S;Piknova B;Schechter AN;Sibmooh N
通讯作者：
Sibmooh N
影响因子：
7.4
作者：
Velmurugan, Shanti;Kapil, Vikas;Ghosh, Suborno M.;Davies, Sheridan;McKnight, Andrew;Aboud, Zainab;Khambata, Rayomand S.;Webb, Andrew J.;Poole, Alastair;Ahluwalia, Amrita
通讯作者：
Ahluwalia, Amrita
影响因子：
158.5
作者：
Becattini, Cecilia;Agnelli, Giancarlo;Prandoni, Paolo
通讯作者：
Prandoni, Paolo
影响因子：
3.3
作者：
Naseem, Khalid M.;Roberts, Wayne
通讯作者：
Roberts, Wayne
影响因子：
4.8
作者：
Nagababu, E;Ramasamy, S;Rifkind, JM
通讯作者：
Rifkind, JM