Dobson G P, Cieslar J H
Department of Physiology and Pharmacology, School of Biomedical and Molecular Sciences, Townsville, Queensland, 4811, Australia.
J Mol Cell Cardiol. 1997 Dec;29(12):3357-63. doi: 10.1006/jmcc.1997.0560.
The aim of this study was to determine tissue water spaces in rat heart in vivo using [14C]-inulin or D-[14C]-mannitol as extracellular markers, and 2,3DPG as whole blood tissue marker. Tracer was injected into the jugular vein of anaesthetised male Sprague-Dawley rats (60 mg/kg) and allowed to equilibrate for 30 min. A blood sample was then taken and the heart was freeze-clamped at liquid N2 temperatures within 5 s of opening the chest cavity. Extracellular space was calculated by dividing the total tissue counts (d/min/g) by plasma counts (d/min/g). The extracellular inulin and mannitol spaces were 0.212+/-0.010 and 0.236+/-0.012 ml/g wet weight tissue, respectively (n=5). Expressed as a percentage of total tissue water (0.79+/-0.012 ml/g wet weight tissue, n=9), the intracellular space was found to be 73.1 (inulin) and 70.1% (mannitol). The interstitial space was calculated by subtracting the counts in tissue due to whole blood from total tissue counts and dividing by plasma counts. The interstitial space was 19.4 and 21. 9%; and the plasma space was 7.5 and 8.0% for the inulin and mannitol methods, respectively. We conclude that (i) there was no significant differences in the inulin or mannitol spaces in the in vivo heart (P<0.05), demonstrating that either marker provides a reliable and quantitative estimate of extracellular space, and (ii) 2,3-DPG can be used as a reliable indicator of tissue whole blood which can then be used in combination with the tracer method to calculate the interstitial and plasma spaces.
本研究的目的是使用[14C]-菊粉或D-[14C]-甘露醇作为细胞外标志物,以及2,3 -二磷酸甘油酸(2,3DPG)作为全血组织标志物,在体内测定大鼠心脏的组织水间隙。将示踪剂注入麻醉的雄性Sprague-Dawley大鼠的颈静脉(60 mg/kg),并使其平衡30分钟。然后采集血样,并在打开胸腔后5秒内于液氮温度下将心脏冷冻钳夹。细胞外间隙通过将总组织计数(每分钟衰变数/克)除以血浆计数(每分钟衰变数/克)来计算。细胞外菊粉和甘露醇间隙分别为0.212±0.010和0.236±0.012 ml/克湿重组织(n = 5)。以总组织水(0.79±0.012 ml/克湿重组织,n = 9)的百分比表示,细胞内间隙分别为73.1%(菊粉)和70.1%(甘露醇)。间质间隙通过从总组织计数中减去全血在组织中的计数,再除以血浆计数来计算。菊粉和甘露醇方法的间质间隙分别为19.4%和21.9%;血浆间隙分别为7.5%和8.0%。我们得出结论:(i)体内心脏的菊粉或甘露醇间隙无显著差异(P<0.05),表明这两种标志物均可提供细胞外间隙的可靠定量估计;(ii)2,3 -二磷酸甘油酸可作为组织全血的可靠指标,然后可与示踪剂方法结合使用来计算间质和血浆间隙。