动脉微栓塞组织病理及血栓调节蛋白变化的实验研究

Changes of histochemical pathology and thrombomodulin in arterial microembolisation

本研究从临床角度出发，对血栓溶解后致小动脉及微动脉栓塞现象进行了研究分析。实验分析研究动脉微栓塞中纤维蛋白成分的免疫组化改变；观察动脉微栓塞后组织灌注的变化趋势：研究动脉微栓塞后组织血栓调节蛋白表达的变化。得出以下结论：（1）纤维蛋白是动脉微栓塞的主要成分。随微栓塞时间的延长，纤维蛋白含量增加。（2）微栓塞初期血流呈现先下降再上升的双相血流现象，随后进行性下降。（3）微栓塞、大栓塞及正常对照组中TM的表达呈显著性差异，微栓塞组织中TM表达显著增多。该研究为动脉微栓塞的诊断、治疗进行了有益的探索。

Objective （ 1 ） Microembolization is one of the pathogenesis of “no- reflow phenomenon” after PCI. In our study, to maintain that fibrin is one of the important ingredients of microemboli which involves in the formation of microembolization soon alter thrombolysis, and thus build up a theoretical basis for microthrombolytic therapy is the main purpose. （2）Analysize the trend of TBF after microembolisation. （3） Comparing the expression of thrombomodulin among mieroembolisation group, embolisation group and the control group.Methods Rat cremasteric arteries were embolized through photochemistry, and mierothomboli formed after thombolysis by staphylokinase. 60 adult male SD mice weighed （180±10） g were randomized separated .30 mice were separated to 5 groups： 5 min, 30 min, 6 0 min, 90 min after microthrombolization and the control group. First, TBF of each group was measured. Then, cremasters were cut down and stored in formaldehyde solution for Masson staining and immunohistochemical analysis. The other 30 adult male SD mice were randomized separated to 3 groups： 90 min after thrombolization, 90min after microthrombolization, and the control group. 5 cremasters of each group were stored in formaldehyde solution for immunohistochemitry.5 cremasters of each group were stored in liquid nitrogen for western blot. Results （ 1 ）Fibrin was observed in pathologic sections as well as fibrinolytic markers t-PA and PAI in immunochemical sections. The more time passed, the more t-PA were found reduced while PAI increased. （2）TBF of 5 rain after microembolisation decreased significantly compared to the control group （P =0.00）. While, TBF of 30 min after microembolisation increased significantly compared to 5min after microembolisation （P =0.00）. TBF of 60min after microembolisation decreased significantly compared to 30 min （P = 0.00）. The same result was found when compared 90min after microembolisation to 60 min after microembolisation （P =0.00）. When TBF measured with Laser Doppler Perfusion lmager, it could be seen that blue and green area expanded 5 min after microembolisation. On the contrary, red area expanded 30min after microembolisation. And TBF of 60 min, 90min after microembolisation witnessed blue and green area progressively decreased. （3）With the formation of thrombolisation, the content of thrombomodulin increased. In immunohistochemical section, the bully area increased.In X-ray picture of Western blot, thrombomodulin expressed in molecular weight 39KD and 46 KD. It can be found that thrombomodulin increased significantly in microthrombolization group in contrast to thrombolization group （P 〈0.05） and the control （P 〈 0.05）.Conclusions （ 1 ） Fibrin is one of the important parts of microthromboli soon after thrombolysis. And the number of fibrous emboli is increasing after microembolization. （2）TBF after microembolisation decreases in the first place. Then, increasing shortly after decreasing. After a short period of time, TBF progressively decrease. （3） Thrombomodulin expressed mostly in microembolisation group compared to embolisation group and the control group.