序贯应用四种不同颜色的非放射性彩色微球测定器官血流量

Measuremeut of organ blood flow using 4 different non-radiated dye extraction colored microspheres in rat abdominal hypertension models

目的探讨以4种颜色的非放射性彩色微球（粉、黄、自、紫）序贯测定同一种动物经历4种不同腹腔内压状态时的脏器血流量的可行性及准确性。方法雄性Wistar大鼠6只，通过腹腔内加压气囊建立0、5、10、15mmHg（1mmHg=0．133kPa）等4种腹腔内压状态，分别以IAPx（x=0、5、10、15）表示。每次持续加压10min，即通过左心室导管注入相应地注入粉色、黄色、白色、紫色中的一色微球。抽取参比血样后立即输注同等量的同种异体血。腹压去除后，间隔15rain。每只大鼠均依次经历4种不同的腹压状态。实验结束，静脉注入饱和氯化钾处死大鼠，并收集组织标本。标本经过碱性组织液消化、超声细胞破碎仪破碎、DMF染料提取及吸光度值测定。依照组织血流量（Qt）[ml／（min·g）]：组织吸光度值（At）×参比血样的血流量（Qb）／参比血样吸光度值（Ab）公式，用以计算脏器血流量。结果微球于血液内分布均一，4组不同腹腔内压下左、右肾之间血流量差异无统计学意义（F=0．177，P〉0．05），[左肾血流量：（2．02±0．47）、（1．39±0．62）、（0．81±0．24）、（0．22±0．09）mr／（min·g）；右肾血流量：（2．23±0．26）、（1．38±0．61）、（0．89±0．31）、（0．24±0．09）mL／（min·g）]具有明显相关（P〈0．01，R=0．687）。随腹腔内压力的增加，肝脏等腹腔内脏器血流量下降明显[肝动脉血流量：（0．16±0．05）、（0．15±0．03）、（0．10±0．03）、（0．02±0．01）ml／（min·g）]；门静脉血流量：（1．73±0．41）、（1．19±0．21）、（0．83±0．18）、（0．38±0．08）ml／（min·g）]。而心、脑等重要脏器血流量在腹腔内压力处于一定范围内（5—10mmHg），能够在一定程度上得以维持，当腹腔内压力升至15mmHg时，心、脑等脏器血流量亦表现为下降趋势[心脏血流量：（2．56±0．95）、（1．69±0．95）、（1．44±0．41）、（0．71±0．24）ml／（min·g）；大脑血流量I（0．49±0．09）、（0．22±0．07）、（0．22±0．06）、（0．19±0．02）ml／（min·g）。微球回收率为（92．0±7．1）％。结论非放射性彩色微球能够完成测定4种不同腹压状态下大鼠多脏器血流量改变，是一种准确可行的血流动力学检测方法，可用于研究同一动物经历不同状态后血流动力学的变化，对于小型动物尤为适用。

Objective To investigate the feasibility and accuracy of the using 4 different colored on-radiated dye extraction colored microspheres to measure multiple organs blood flow in one rat, which ustained four conditions of intra-abdominal pressuring. Methods An inflatable gasbag was inserted into bdominal cavity of rat, which was used to create the model of non-pneumatic intra-abdominal pressure IAPx） [x =0, 5, 10, 15 mm Hg （ 1 mm Hg =0. 133 kPa） ]. Each condition of IAPx sustained 10 min. One of the four non-radiated colored dyed microspheres was infused through cannulatian tube in the left mtricle in the sequence of pink, yellow, white and purple. After drawing reference blood sample, comlementay blood was transfused in an equal amount from another transfusion rat. The interval time was 15 in. All six rats were suffered from the four conditions of intra-abdominal pressure. At the end of the exeriment, animals were killed by intravenous injection of saturated potassium chloride solution, and aimed gan samples were harvested. The organ blood flows were calculated after a sequence of dealing with sames, such as alkaline digesting, ultrasonography cell crashing, DMF dying extraction, and the dye concenftion was quantified by spectrophotometry. Tissue blood flow was calculated by an equation： Organ blood ,w = [ AU per gram of tissue ] x [ reference withdrawal rate ] / [ AU per gram of reference blood ]. Results 1 animals were endured the four conditions （0, 5, 10, 15 mmHg） of intra-abdominal pressure. The disbution of microspheres in the blood flow was uniform, and the blood flow through the right kidney and left kidney was matched equally： for left kidney [ （2.02 ±0. 47）, （ 1.39 ±0. 62）, （0. 81 ±0. 24）, （0. 22 ±0.09） ml·（min·g）]; For right kidney [（2.23 ±0.26）, （1.38 ±0.61）, （0.89 ±0.31）, （0.24±0. 09） ml/（min.g） ,F = 0. 177 ,P 〉 0. 05 ], and had close correlation （ P 〈 0. 01 ,R = 0. 687 ）. With the increase of the abdominal pressure from 5 to 10 mmHg, the blood flow of abdominal organs （such as liver） was decreased significantly [Hepatic artery： （0. 16 ±0. 05）, （0. 15 ±0. 03）, （0.10 ±0. 03）, （0. 02 ±0.01） ml·（min·g） ; Portal vein： （1.73 ±0.41）, （1.19 ±0.21）, （0. 83 ±0. 18）, （0.38 ±0.08） ml/ （ min ·g） ]. On the other hand, the blood flow of the vital organs （ such as heart and brain） was maintained in the abdominal pressure of 5-10 mm Hg. When the abdominal pressure was increased to 15 mmHg, the blood floow of the vital organs showed the decreased tendency [ Heart ： （2. 56 ±0. 95 ）, （ 1.69 ±0. 95 ）, （1.44 ±0.41）, （0.71 ±0.24） ml/（min·g） ; Brain： （0.49 ±0.09）, （0.22 ±0.07）, （0.22 ±0.06）, （0. 19 ±0. 02） ml/（ min· g） ]. The recovery rate of microspheres was （ 92. 0 ±7. 1 ） %. Conclusion The use of dye extraction colored microspheres should allow to undergo hemodynamic study on different organ blood flow in one animal. The method is feasible and accurate, especially suitable for small animals.