兔深静脉血栓形成后内皮细胞凋亡变化

    Apoptosis of endothelial cells after deep venous thrombosis in rabbits

    • 摘要: 目的: 探讨兔急性深静脉血栓形成后血管内皮细胞的凋亡及其在病理过程中的意义。方法: 55只健康家兔随机分为血栓组25只、假手术组25只和正常组5只。血栓组家兔采用开腹手术结扎肾下段后腔静脉建立血栓模型,假手术组模拟手术和正常组直接取材作对照。3组对照分别于实验后第12h及1、2、4、7天取实验段血管。采用透射电镜观察内皮细胞凋亡情况,应用TUNEL法和免疫组织化学方法检测血管内皮细胞凋亡和相关基因bax、bcl-2蛋白的表达。结果: 血栓组1、2、4天透射电镜下见内皮细胞凋亡。TUNEL法标记阳性表达在术后1、2天达高峰,两者差异无统计学意义(P>0.05),并于4、7天降低,不同时相血栓组与假手术组间差异有统计学意义(P<0.05~P<0.01)。血栓组bcl-2的表达进行性升高,术后2天达到高峰,4、7天有所下降,不同时相血栓组与假手术组间差异有统计学意义(P<0.05~P<0.01)。结论: 家兔急性静脉血栓形成后1~2天内皮细胞凋亡达到高峰,然后进行性下降。bax与bcl-2基因参与血栓形成后血管内皮细胞凋亡的调控。

       

      Abstract: Objective: To explore the apoptosis of endothelial cells after acute deep venous thrombosis in rabbit and its significanse in the pathological processes. Methods: Fifty-five adult rabbits were randomly divided into the normal group (n=5) in which the samples were gotten directly, the thrombosis group (TB, n=25) caused by ligating a sect of the posterior vena cava under renal vein to induce thrombosis model, the sham operation group (SH, n=25) that were simply belly open and close to be used as controls. In different periods after acute deep venous thrombosis, morphological change of the endothelial cell apoptosis on target vessels was observed by means of transmission electron microscopy. In situ end-labeling of DNA fragments (TUNEL) was used to detect the endothelial cell apoptosis. Bax and bcl-2 proteins expressed by endothelial cells were determined with immunohistochemical staining. Results: Venous thrombosis models were successfully obtained in all cases. Transmission electron microscopy results showed that apoptosis of partial endothelial cells on target vessels on the 1st day, the 2nd day and the 4th day. TUNEL-positive cell rate reached the maximum on the first day, the second day and there was no significant difference between the two groups (P>0.05), and decreased gradually on the 4th day and the 7th day in TB group. There was significant difference between TB group and SH group respective time (P<0.05-P<0.01). Expression of bcl-2 protein in TB group was increased gradually with the days passing by while bax protein reached the peak on 2nd days and decreased gradually 4th day, 7th day, there was marked difference between TB group and SH group at different time (P<0.05-P<0.01). Conclusions: The apoptosis of the endothelial cells after acute deep venous thrombosis occurs, reaches the peak on the first and the second day and decreases gradually after 2nd days. The bcl-2 and bax participate in the regulation and control of the apoptosis of the endothelial cells.

       

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