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目的研究同位素~(99)Tc~m示踪血栓技术在家兔脑卒中模型上的应用,并动态示踪观察重组组织型纤溶酶原激活剂(rt-PA)对血栓的溶解效果。方法将新鲜洗脱的放射性高锝酸钠(5 m Ci/2 mL,92.5 MBq/mL)0.5 mL与5 mg/mL氯化亚锡30μL充分混合成示踪标记液;分别向全血、红细胞、血浆中加入20μL示踪标记液进行同位素标记,加入50μL CaCl_2(0.5 mol/L)与牛凝血酶(50 IU/mL)混合物,迅速吸入聚乙烯塑料管(PE80)中,37℃固化2 h后取出血栓,分割成10 mm的示踪血栓,并计算标记率。采用经兔颈外动脉逆向颈内动脉插管方法,注射示踪血栓栓塞大脑中动脉,制备脑卒中模型,用γ计数仪在兔头部测定放射性强度即每分钟计数(Counts Per Minute,CPM)的动态变化,并观察临床等效剂量工具药rt-PA4.5 mg/kg的溶栓效果。结果全血血栓标记率为53.5%,红细胞与血浆标记比率为4.5∶1,家兔大脑中动脉血栓栓塞后CPM明显增加,约为本底的5.1±1.3倍,表明模型制备成功,检测可靠。静脉给予工具药rt-PA能产生显著的渐进性溶栓效果。结论锝标记主要以标记红细胞为主,对血浆及纤维蛋白原影响较小。~(99)Tc~m示踪血栓技术制备的脑卒中模型,分离区明显,模型制备成功、指标检测科学,模型对药物疗效应答能力的反应稳定、可靠。
Objective To investigate the application of isotope 99 Tc m trace thrombosis in stroke model in rabbits and to observe dynamically the effect of recombinant tissue plasminogen activator (rt-PA) on the thrombosis. Methods 0.5 mL of freshly washed radioactive sodium pertechnetate (5 m Ci / 2 mL, 92.5 MBq / mL) and 30 μL of 5 mg / mL stannous chloride were mixed thoroughly into a labeled solution. The whole blood, erythrocytes , 20μL tracer was added to the plasma for isotope labeling. Fifty microliters CaCl2 (0.5 mol / L) and bovine thrombin (50 IU / mL) mixture were added to the tube and rapidly inoculated into polyethylene plastic tube (PE80) Thrombus was removed and divided into 10 mm tracer thrombus, and the labeling rate was calculated. The model of stroke was established by injecting the middle cerebral artery of thromboembolism by transcatheter arterial cannulation of rabbit external carotid artery. Counts Per Minute (CPM) The dynamic changes of thrombolytic activity were observed and the clinical thrombolysis effect of rt-PA4.5 mg / kg was observed. Results The rate of whole blood thrombosis was 53.5%. The ratio of erythrocyte to plasma was 4.5:1. The CPM of middle cerebral artery thromboembolism in rabbits increased significantly, which was about 5.1 ± 1.3 times of the background, indicating that the model was successfully prepared and the detection was reliable. Intravenous administration of the drug rt-PA produced a significant gradual thrombolytic effect. Conclusion The majority of labeled technetium labeled red blood cells, plasma and fibrinogen less affected. ~ (99) Tc ~ m tracer model of thrombosis prepared stroke model, the separation area was obvious, the model was prepared successfully, the index of scientific testing, the model response to the drug response to the efficacy of stable and reliable.