The multiple-indicator dilution technique for characterization of normal and retrograde flow in once-through rat liver perfusions.
The multiple-indicator dilution technique for characterization of normal and retrograde flow in once-through rat liver perfusions.
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用于表征一次性大鼠肝脏灌注中正常和逆行血流的多指示剂稀释技术。
DOI：
10.1002/hep.1840090221
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Pang,KS
中科院分区：
文献类型：
--
作者：
St-Pierre,MV;Schwab,AJ;Goresky,CA;Lee,WF;Pang,KS
正常和逆行大鼠肝脏灌注技术已被广泛用于探测药物代谢活动的区域差异。这种方法的有效性要求在正常和逆行灌注期间基质进入相同的组织空间。使用多指示剂稀释技术，我们目前研究的程度逆行灌注改变的空间可访问的非消除的参考。将~（51）Cr标记的红细胞、~（125）I-白蛋白、~（14）C-蔗糖和~（3 H2 O）分别注入大鼠门静脉（正常）或肝静脉（逆行），灌注速度为10 ml/min。在220秒内连续收集流出灌注液，以表征标签的通过时间和分布空间。在逆行灌注期间，红细胞，白蛋白和蔗糖曲线达到峰值后，低于正常灌注期间，而水曲线相似。红细胞、白蛋白和蔗糖的通过时间延长（p< 0.005），而水的通过时间没有变化。因此，逆行血流导致显著更大的窦血容量（45%），白蛋白Disse空间（42%）和蔗糖Disse空间（25%）比在正常流动，而总的和细胞内水的分布空间保持不变。通过电子显微镜证实了血管树的扩张，偶尔观察到细胞间凹和Disse间隙变宽的孤立病灶。细胞超微结构没有变化，正常和逆行灌注之间的胆汁流速，AST释放，灌注压，耗氧量和乙醇的代谢清除没有差异，乙醇是一种具有流动限制分布的底物，可迅速与细胞水平衡（肝提取率几乎相同：正常与逆行，在6至7.4 mM输入浓度下为0.50与0.48）。这些研究结果表明，肝脏的功能和代谢能力在逆行灌注过程中保持不受干扰，使该技术适合于药物代谢酶的区域差异的调查。
The technique of normal and retrograde rat liver perfusion has been widely used to probe zonal differences in drug-metabolizing activities. The validity of this approach mandates the same tissue spaces being accessed by substrates during both normal and retrograde perfusions. Using the multiple-indicator dilution technique, we presently examine the extent to which retrograde perfusion alters the spaces accessible to noneliminated references. A bolus does of 51 Cr-labeled red blood cells, 125 I-albumin, 14 C-sucrose and 3 H 2 O was injected into the portal (normal) or hepatic (retrograde) vein of rat livers perfused at 10 ml per min per liver. The outflow perfusate was serially collected over 220 sec to characterize the transit times and the distribution spaces of the labels. During retrograde perfusion, red blood cells, albumin and sucrose profiles peaked later and lower than during normal perfusion, whereas the water curves were similar. The transit times of red blood cells, albumin and sucrose were longer (p< 0.005), whereas those for water did not change. Consequently, retrograde flow resulted in significantly larger sinusoidal blood volumes (45%), albumin Disse space (42%) and sucrose Disse space (25%) than during normal flow, whereas the distribution spaces for total and intracellular water remained unaltered. The distension of the vascular tree was confirmed by electron microscopy, by which occasional isolated foci of widened intercellular recesses and spaces of Disse were observed. Cellular ultrastructure was otherwise unchanged, and there was no difference found between normal and retrograde perfusion for bile flow rates, AST release, perfusion pressure, oxygen consumption and metabolic removal of ethanol, a substrate with flow-limited distribution, which equilibrates rapidly with cell water (hepatic extraction ratios were virtually identical: normal vs. retrograde, 0.50 vs. 0.48 at 6 to 7.4 mM input concentration). These findings suggest that the functional and metabolic capacities of the liver remain unperturbed during retrograde perfusion, rendering the technique suitable for the investigation of zonal differences in drug-metabolizing enzymes.