血浆腺苷浓度的测量：方法学与生理学考量

Measurement of plasma adenosine concentration: methodological and physiological considerations.

作者信息

Gewirtz H, Brown P, Most A S

出版信息

Proc Soc Exp Biol Med. 1987 May;185(1):93-100. doi: 10.3181/00379727-185-42523.

Abstract

This study tested the hypothesis that measurements of plasma adenosine concentration made on samples of blood obtained in dipyridamole and EHNA (i.e., "stopping solution") may be falsely elevated as a result of ongoing in vitro production and accumulation of adenosine during sample processing. Studies were performed with samples of anticoagulated blood obtained from anesthesized domestic swine. Adenosine concentration of ultra filtrated plasma was determined by high-pressure liquid chromatography (HPLC). The following parameters were evaluated: (i) rate of clearance of [3H]adenosine added to plasma, (ii) endogenous adenosine concentration of matched blood samples obtained in "stopping solution" alone, "stopping solution" plus EDTA, and perchloric acid (PCA), (iii) plasma and erythrocyte endogenous adenosine concentration in nonhemolyzed samples, and (iv) plasma adenosine concentration of samples hemolyzed in the presence of "stopping solution" alone or "stopping solution" plus EDTA. We observed that (i) greater than or equal to 95% of [3H]adenosine added to plasma is removed from it by formed elements of the blood in less than 20 s, (ii) plasma adenosine concentration of samples obtained in "stopping solution" alone is generally 10-fold greater than that of matched samples obtained in "stopping solution" plus EDTA, (iii) deliberate mechanical hemolysis of blood samples obtained in "stopping solution" alone resulted in substantial augmentation of plasma adenosine levels in comparison with matched nonhemolyzed specimens--addition of EDTA to "stopping solution" prevented this, and (iv) adenosine content of blood samples obtained in PCA agreed closely with the sum of plasma and erythrocyte adenosine content of samples obtained in "stopping solution" plus EDTA. The data obtained demonstrate that (i) plasma adenosine concentrations are falsely elevated in samples of blood obtained in "stopping solution" alone, and (ii) addition of EDTA to "stopping solution" blocks in vitro production and accumulation of adenosine. Finally, rapid removal of adenosine from plasma by formed elements of blood may make it difficult to employ measurements of plasma adenosine concentration to assess physiological processes even in the absence of in vitro production of the nucleoside.

摘要

本研究检验了以下假设

在双嘧达莫和EHNA（即“终止溶液”）中采集的血样所测得的血浆腺苷浓度，可能会因样本处理过程中腺苷的持续体外生成和积累而被错误地升高。研究使用了从麻醉的家猪采集的抗凝血液样本。通过高压液相色谱法（HPLC）测定超滤血浆中的腺苷浓度。评估了以下参数：（i）添加到血浆中的[³H]腺苷的清除率，（ii）仅在“终止溶液”、“终止溶液”加乙二胺四乙酸（EDTA）和高氯酸（PCA）中采集的匹配血样的内源性腺苷浓度，（iii）未溶血样本中的血浆和红细胞内源性腺苷浓度，以及（iv）仅在“终止溶液”或“终止溶液”加EDTA存在下溶血的样本的血浆腺苷浓度。我们观察到：（i）添加到血浆中的[³H]腺苷在不到20秒内有大于或等于95%被血液中的有形成分从血浆中清除，（ii）仅在“终止溶液”中采集的样本的血浆腺苷浓度通常比在“终止溶液”加EDTA中采集的匹配样本高10倍，（iii）仅在“终止溶液”中采集的血样经故意机械溶血后，与匹配的未溶血样本相比，血浆腺苷水平大幅升高——在“终止溶液”中添加EDTA可防止这种情况，以及（iv）在PCA中采集的血样的腺苷含量与在“终止溶液”加EDTA中采集的样本的血浆和红细胞腺苷含量之和密切相符。所获得的数据表明：（i）仅在“终止溶液”中采集的血样的血浆腺苷浓度被错误地升高，以及（ii）在“终止溶液”中添加EDTA可阻断腺苷的体外生成和积累。最后，血液中的有形成分可迅速从血浆中清除腺苷，这可能使得即使在没有核苷体外生成的情况下，利用血浆腺苷浓度测量来评估生理过程也变得困难。