血浆中腺苷二磷酸及相关物质的检测与测定

Detection and determination of adenosine diphosphate and related substances in plasma.

作者信息

Ireland D M, Mills D C

出版信息

Biochem J. 1966 May;99(2):283-96. doi: 10.1042/bj0990283.

DOI:10.1042/bj0990283

PMID:5947146

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1264995/

Abstract

A method is described for detecting and determining the products of metabolism of ADP added to plasma at initial concentrations of about 1mum-ADP. 2. ATP, ADP, AMP, adenosine, inosine and hypoxanthine were detected in human platelet-rich plasma after incubation with ADP and in the presence of either heparin or heparin-citrate. 3. The products of incubation of ADP with human platelet-poor plasma in the presence of heparin were the same as with platelet-rich plasma, except that, when the initial concentration of ADP was 1.5mum, little or no ATP was detected. 4. The ATP detected in platelet-rich plasma when 1.5mum-ADP was initially incubated was present in the platelets and not in the plasma. 5. The time for 50% decay of ADP in either platelet-rich or platelet-poor plasma in the presence of heparin was about 20min. when the initial concentration of ADP was 200mum, but was 6-9min. when the initial ADP concentration was 1.5-2.5mum. The corresponding values in the presence of heparin-citrate were about 45min. and about 9-12min. respectively. 6. Hypoxanthine accumulated to a greater extent in platelet-rich than in platelet-poor plasma after the addition of ADP. 7. After incubation for 15-20min. of either platelet-rich plasma or suspensions of washed platelets in saline with adenosine at an initial concentration of about 3-4mum, ATP, ADP and AMP were detected in the platelets. Similar incubations of washed platelets with inosine also showed the formation of these substances, but to a much less extent. 8. After the addition of adenosine to suspensions of washed platelets in saline, inosine and hypoxanthine were detected in the incubation mixture. After the addition of inosine, hypoxanthine was detected. 9. When ADP at an initial concentration of 1.5mum was added to platelet-rich plasma containing adenosine deaminase, no adenosine was detected in the incubation mixture. There was no difference in the rate of decay of ADP in the presence or absence of the deaminase, but ATP formation was decreased in its presence.

摘要

本文描述了一种用于检测和测定初始浓度约为1μM - ADP的添加到血浆中的ADP代谢产物的方法。2. 在与ADP孵育后，且存在肝素或肝素 - 柠檬酸盐的情况下，在富含人血小板的血浆中检测到了ATP、ADP、AMP、腺苷、肌苷和次黄嘌呤。3. 在肝素存在下，ADP与人贫血小板血浆孵育的产物与富含血小板血浆的相同，只是当ADP的初始浓度为1.5μM时，几乎检测不到或未检测到ATP。4. 当最初孵育1.5μM - ADP时，在富含血小板血浆中检测到的ATP存在于血小板中而非血浆中。5. 在肝素存在下，富含血小板或贫血小板血浆中ADP 50%衰减的时间，当ADP初始浓度为200μM时约为20分钟，但当ADP初始浓度为1.5 - 2.5μM时为6 - 9分钟。在肝素 - 柠檬酸盐存在下的相应值分别约为45分钟和约9 - 12分钟。6. 添加ADP后，富含血小板血浆中次黄嘌呤的积累程度比贫血小板血浆中更大。7. 在富含血小板血浆或用初始浓度约为3 - 4μM的腺苷在盐水中洗涤的血小板悬液孵育15 - 20分钟后，在血小板中检测到了ATP、ADP和AMP。用肌苷对洗涤的血小板进行类似孵育也显示了这些物质的形成，但程度要小得多。8. 向盐水中洗涤的血小板悬液中添加腺苷后，在孵育混合物中检测到了肌苷和次黄嘌呤。添加肌苷后，检测到了次黄嘌呤。9. 当将初始浓度为1.5μM的ADP添加到含有腺苷脱氨酶的富含血小板血浆中时，在孵育混合物中未检测到腺苷。存在或不存在脱氨酶时ADP的衰减速率没有差异，但存在脱氨酶时ATP的形成减少。