Determination of carbon monoxide (CO) in rodent tissue: Effect of heme administration and environmental CO exposure
Determination of carbon monoxide (CO) in rodent tissue: Effect of heme administration and environmental CO exposure
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DOI：
10.1016/j.ab.2005.03.019
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发表时间：
2005-06-15
影响因子：
2.9
通讯作者：
Stevenson, DK
中科院分区：
文献类型：
--
作者：
Vreman, HJ;Wong, RJ;Stevenson, DK
在血红素降解过程中内源性产生的一氧化碳（CO）被认为是血管和神经组织中的信使分子。为了研究这一作用，重要的是要确定CO浓度在目标组织前后的扰动。在这里，我们描述了一个敏感和可重复的方法，这是线性的和准确的，并提供了一些例子，其应用于定量组织CO浓度前后的扰动。成年大鼠和小鼠的组织超声（20% w/w），体积为0.04-8 mg鲜重（FW），与磺基水杨酸在0℃下孵育>= 30 min。通过气相色谱法对释放到顶空的CO进行定量分析。组织CO浓度(意思是+ / - SD, pmol / mg FW)如下:血液(47 + / - 10,45 + / - 5),肌肉(4 + / - 4,10 + / - 1)、肾(5 + / - 2,7 + / - 2)、心(6 + / - 3 6 + / - 1)、脾(11 + / - 3,6 + / - 1),肝脏(4 + / - 1,5 + / - 1),小肠(2 + / - 1,4 + / - 2)、肺(2 + / - 1,3 + / - 1),睾丸(1 + / - 1,2 + / - 1),和大脑(2 + / - 1,2 + / - 0)在未经处理的大鼠(n = 3)和鼠标(n 5),分别。在大鼠和小鼠之间，只有肌肉和脾脏中的CO浓度有显著差异(p
Carbon monoxide (CO), produced endogenously during heme degradation, is considered a messenger molecule in vascular and neurologic tissues. To study this role, it is important to determine CO concentration in target tissues pre- and post-perturbations. Here, we describe a sensitive and reproducible method, which is linear and accurate, and provide some examples of its application for quantitation of CO concentrations in tissues pre- and post-perturbations. Tissues from adult rats and mice were sonicated (20% w/w), and volumes representing 0.04-8 mg fresh weight (FW) were incubated at 0 degrees C for >= 30 min with sulfosalicylic acid. CO liberated into the headspace was quantitated by gas chromatography. Tissue CO concentrations (mean +/- SD, pmol CO/mg FW) were as follows: blood (47 +/- 10, 45 +/- 5), muscle (4 +/- 4, 10 +/- 1), kidney (5 +/- 2, 7 +/- 2), heart (6 +/- 3 6 +/- 1), spleen (11 +/- 3, 6 +/- 1), liver (4 +/- 1, 5 +/- 1), intestine (2 +/- 1, 4 +/- 2), lung (2 +/- 1, 3 +/- 1), testes (1 +/- 1, 2 +/- 1), and brain (2 +/- 1, 2 +/- 0) in untreated rat (n = 3) and mouse (n 5), respectively. Between the rat and the mouse, only CO concentrations in the muscle and spleen were significantly different (p