FALSE IN VITRO AND IN VIVO ELEVATIONS OF URIC ACID LEVELS IN MOUSE BLOOD
FALSE IN VITRO AND IN VIVO ELEVATIONS OF URIC ACID LEVELS IN MOUSE BLOOD
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DOI：
10.1080/15257770.2013.865742
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发表时间：
2014-01-01
影响因子：
1.3
通讯作者：
Hosoyamada, Makoto
中科院分区：
文献类型：
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作者：
Watanabe, Tamaki;Tomioka, Naoko H.;Hosoyamada, Makoto
据报道，小鼠血液中的尿酸 (UA) 水平范围广泛，从 0.1 muM 到 760 muM。本研究的目的是证明小鼠血液中 UA 水平的体外和体内虚假升高。雄性ICR小鼠用戊巴比妥（呼吸小鼠）麻醉或用过量乙醚处死（非呼吸小鼠）。将收集的血液分配到 MiniCollect (R) 管中并在室温下体外孵育 0 或 30 分钟。分离血浆或血清后，使用HPLC测定UA和次黄嘌呤的水平。从未孵化的呼吸小鼠血浆中，体内UA水平的真实值为13.5+/-1.4μM。然而，体外孵化后，小鼠血液中的UA水平增加了3.9倍。采血后小鼠血液中 UA 水平的这种“体外假性升高”受到别嘌呤醇（一种黄嘌呤氧化酶抑制剂）的抑制。黄嘌呤氧化酶将小鼠血清中的次黄嘌呤转化为 UA，而次黄嘌呤是在培养过程中从血细胞中释放出来的。不呼吸小鼠的血浆 UA 水平比呼吸小鼠高 19 倍。采血前 UA 水平的这种“体内假性升高”可通过使用 α-拮抗剂酚妥拉明进行预处理来抑制。乙醚过度麻醉可能引起血管收缩和缺血，从而将细胞内 ATP 降解为 UA。为了准确测量小鼠血液中的 UA 水平，必须在麻醉呼吸小鼠采血后立即分离血浆，以避免体外和体内 UA 水平的错误升高。
Uric acid (UA) levels in mouse blood have been reported to range widely from 0.1 mu M to 760 mu M. The aim of this study was to demonstrate false in vitro and in vivo elevations of UA levels in mouse blood. Male ICR mice were anesthetized with pentobarbital (breathing mice) or sacrificed with overdose ether (non-breathing mice). Collected blood was dispensed into MiniCollect (R) tubes and incubated in vitro for 0 or 30 min at room temperature. After separation of plasma or serum, the levels of UA and hypoxanthine were determined using HPLC. From the non-incubated plasma of breathing mice, the true value of UA level in vivo was 13.5 +/- 1.4 mu M. However, UA levels in mouse blood increased by a factor of 3.9 following incubation in vitro. This "false in vitro elevation" of UA levels in mouse blood after blood sampling was inhibited by allopurinol, a xanthine oxidase inhibitor. Xanthine oxidase was converted to UA in mouse serum from hypoxanthine which was released from blood cells during incubation. Plasma UA levels from non-breathing mice were 19 times higher than those from breathing mice. This "false in vivo elevation" of UA levels before blood sampling was inhibited by pre-treatment with phentolamine, an a-antagonist. Over-anesthesia with ether might induce a-vasoconstriction and ischemia and thus degrade intracellular ATP to UA. For the accurate measurement of UA levels in mouse blood, the false in vitro and in vivo elevations of UA level must be avoided by immediate separation of plasma after blood sampling from anesthetized breathing mice.