大鼠静脉血栓栓塞症凝血—纤溶标志物动态变化的研究

发布时间：2018-05-10 15:14

本文选题：静脉血栓栓塞症 + 可溶性纤维蛋白复合物　；参考：《大连医科大学》2012年硕士论文

【摘要】：研究背景及目的静脉血栓栓塞症（Venous Thromboembolism, VTE）包括深静脉血栓形成（Deep Venous Thrombosis, DVT）和肺血栓栓塞症（PulmonaryThromboembolism, PTE）。DVT和PTE不是两种独立的疾病，而是VTE一个连续过程。临床确诊VTE的方法主要依靠CT肺动脉造影、放射性核素肺通气/血流灌注扫描、磁共振血管造影及肺动脉造影等影像学检查，至今上述检查不仅仍未普及，而且对于重症或特殊体质患者也存在禁忌。因此，寻找一种早期敏感性和特异性高、简便易行且费用低廉的血浆标志物成为关键问题。本研究通过建立大鼠静脉血栓栓塞症模型，动态检测大鼠血浆中凝血-纤溶标记物——可溶性纤维蛋白复合物（SFC）、凝血因子ⅩⅢ (FⅩⅢ)和D-二聚体(D-dimer)的水平变化，深入理解VTE的病理生理过程以及上述凝血-纤溶标记物的作用和意义。材料和方法Sprague-Dawley大鼠18只，雌雄各半，随机分为DVT组、DVT-PTE组、假手术（Sham）组，每组6只。用微型血管夹阻滞左侧股静脉血流制备DVT模型，DVT造模成功后3天取出左侧股静脉栓子，从右侧股静脉注入，制备PTE模型。假手术组只分离左侧股静脉。造模成功后，分别于栓塞前（0h）和栓塞后不同时间点（3hs、6hs、12hs、1d、2ds、3ds）采血，应用酶联免疫吸附法（ELISA法）测定各组栓塞前和栓塞后不同时间点大鼠血浆中SFC、FⅩⅢ和D-dimer的水平，描绘各项标志物的浓度-时间柱形图及浓度-时间线图，应用计量资料以均数±标准差表示，使用配对t检验，对栓塞后和栓塞前的各项指标进行统计学分析，组间分析采用单因素方差分析，P0.05为差异具有统计学意义。结果 1.血浆SFC水平 DVT组：SFC水平在2ds达峰值（19.795±9.626ug/ml），与0h（4.108±0.809ug/ml）比较，差异具有统计学意义（t=2.923，P=0.033），至第3天恢复正常水平。 DVT-PTE组：SFC水平在3hs即达峰值22.643±19.417ug/ml，但与栓塞前（5.192±0.338ug/ml）相比，差异无统计学意义（t=2.213，P=0.078），至第2-3天仍维持较高水平（11.212±12.742ug/ml；11.138±11.839ug/ml，但与栓塞前比较差异无统计学意义（t=1.147，P=0.303；t=1.216，P=0.278）。 Sham组：SFC水平在3h即开始升高（17.816±4.702ug/ml），至12hs时达峰值（24.322±1.101ug/ml），与0h（3.698±0.676ug/ml）比较，差异均具有显著性（t=6.607，P=0.007；t=43.729，P=0.000），至第3天恢复正常水平。 2.血浆FⅩⅢ水平 DVT组：FⅩⅢ水平在6hs达峰值（557.685±473.435ng/ml），与0h（77.563±22.949ng/ml）比较，，差异具有统计学意义（t=3.204，P=0.029），至第3天仍维持较高水平（291.523±206.034ng/ml），但与栓塞前比较差异无统计学意义（t=2.259，P=0.073）。 DVT-PTE组：FⅩⅢ水平在3hs即达峰值（638.716±674.793ng/ml），但与栓塞前（242.180±26.282ng/ml）相比，差异无统计学意义（t=1.449，P=0.207）。 Sham组：FⅩⅢ水平在术后2ds达峰值（554.295±180.036ng/ml），与术前（94.716±13.475ng/ml）相比，差异具有统计学意义（t=5.517，P=0.012），至第3天仍维持较高水平（398.217±214.255ng/ml），但与栓塞前比较差异无统计学意义（t=2.670，P=0.076）。 3.血浆D-dimer水平： DVT组：D-dimer水平在6hs达峰值（353.380±74.235ng/ml），与0h（40.316±4.345ng/ml）相比，差异具有统计学意义（t=2.298，P=0.03），24小时仍维持较高水平（229.159±181.274ng/ml），与0h比较，差异具有显著性（t=2.995，P=0.02），至第3天恢复正常水平。 DVT-PTE组：D-dimer水平在12-24hs升高并达峰值（476.576±317.067ng/ml；481.177±322.763ng/ml），与栓塞前（55.785±10.489ng/ml）比较，差异均具有统计学意义（t=3.18，P=0.025；t=3.159，P=0.025）。至第2-3天仍维持较高水平（382.956±341.929ng/ml，381.943±346.108ng/ml），但与栓塞前比较差异无统计学意义（t=2.361，P=0.065；t=2.327，P=0.067）。 Sham组：D-dimer水平术后3hs即升高（450.881±165.966ng/ml），24hs达高峰（504.488±124.615ng/ml），与术前（51.050±6.795ng/ml）相比，差异均具有显著性（t=6.071，P=0.000；t=9.170，P=0.000）。2-3天恢复正常水平。结论 1.凝血-纤溶标志物——血浆SFC、FⅩⅢ和D-dimer水平的升高是实验性VTE的早期敏感指标，并呈动态变化，但并非特异。 2.DVT和PTE作为VTE的不同阶段，凝血-纤溶标志物的表达不尽相同，DVT是SFC在FⅩⅢ交联作用下血栓形成，并同时被溶解的动态过程；而PTE则以栓子栓塞后不断溶解过程为主，尽管SFC和FⅩⅢ有升高趋势，但与栓塞前比较差异无显著性。 3.手术创伤可导致凝血-纤溶标志物——血浆SFC、FⅩⅢ和D-dimer水平的升高，是VTE的易患因素。
[Abstract]:Background and objective Venous Thromboembolism (VTE) includes deep venous thrombosis (Deep Venous Thrombosis, DVT) and pulmonary thromboembolism (PulmonaryThromboembolism, PTE).DVT and PTE are not two independent diseases, but a continuous process of VTE. Radionuclide pulmonary ventilation / perfusion scan, magnetic resonance angiography and pulmonary arteriography are not only still not popularized, and there are also taboos for patients with severe or special physique. Therefore, looking for a kind of early sensitivity and specificity, easy and inexpensive plasma markers become a close. Key problem.
In this study, a rat model of venous thromboembolism was established to dynamically detect the levels of coagulation fibrinolytic markers, soluble fibrin complex (SFC), coagulation factor III (F III) and D- two polymer (D-dimer) in rat plasma, and to understand the pathophysiological process of VTE and the role and meaning of the above coagulation fibrinolytic markers. Righteousness.
Materials and methods 18 Sprague-Dawley rats, male and female, were divided randomly into group DVT, group DVT-PTE, sham operation (Sham) group, 6 in each group. DVT model was prepared by blocking the left femoral vein blood flow with microvascular clamp. The left femoral vein embolus was taken out 3 days after the success of DVT modeling, and the PTE model was prepared from the right femoral vein. The sham operation group only separated the left femoral statics. Blood samples were collected at different time points before embolism (0h) and after embolization (3hs, 6HS, 12hs, 1D, 2ds, 3ds) after embolization. The levels of SFC, F, III and D-dimer were measured by enzyme linked immunosorbent assay (ELISA method) in each group of rats before and after embolization. The concentration time column diagram and concentration time line were depicted. The measurement data were expressed with mean standard deviation, and paired t test was used to analyze the indexes of post embolism and pre embolism. The analysis between groups was analyzed by single factor analysis of variance, and the difference of P0.05 was statistically significant.
Result
1. plasma SFC level
Group DVT: the level of SFC was at the peak of 2ds (19.795 + 9.626ug/ml). Compared with 0h (4.108 + 0.809ug/ml), the difference was statistically significant (t=2.923, P=0.033), and the normal level was restored to third days.
Group DVT-PTE: the level of SFC was 22.643 + 19.417ug/ml at 3hs, but there was no statistically significant difference (t=2.213, P=0.078) compared with before embolization (t=2.213, P=0.078), and still maintained a high level (11.212 + 12.742ug/ml, 11.138 + 11.839ug/ml) on day 2-3, but there was no significant difference from before embolism (t=1.147, P=0.303; t=1.216,).
Group Sham: the level of SFC began to rise at 3H (17.816 + 4.702ug/ml) and reached a peak (24.322 + 1.101ug/ml) at 12hs. Compared with 0h (3.698 + 0.676ug/ml), the difference was significant (t=6.607, P=0.007; t=43.729, P=0.000), and recovered to the normal level to third days.
2. plasma F level III level
Group DVT: the level of F III was peak at 6HS (557.685 + 473.435ng/ml). Compared with 0h (77.563 + 22.949ng/ml), the difference was statistically significant (t=3.204, P=0.029), and still maintained a high level (291.523 + 206.034ng/ml) to third days, but there was no statistical significance (t=2.259, P=0.073) before the embolism.
Group DVT-PTE: the level of F III was peak at 3hs (638.716 + 674.793ng/ml), but there was no statistically significant difference compared with that before embolization (242.180 + 26.282ng/ml) (t=1.449, P=0.207).
In group Sham, the peak value of F III was peak (554.295 + 180.036ng/ml) after operation. Compared with preoperative (94.716 + 13.475ng/ml), the difference was statistically significant (t=5.517, P=0.012), and still maintained a high level (398.217 + 214.255ng/ml) to third days, but there was no statistical difference compared with before embolization (t=2.670, P=0.076).
3. plasma D-dimer level:
Group DVT: the peak value of D-dimer was at the peak of 6HS (353.380 + 74.235ng/ml). Compared with 0h (40.316 + 4.345ng/ml), the difference was statistically significant (t=2.298, P=0.03) and still maintained a high level (229.159 + 181.274ng/ml) for 24 hours. Compared with 0h, the difference was significant (t= 2.995, P=0.02), and recovered to the normal level to third days.
Group DVT-PTE: the level of D-dimer increased at 12-24hs and reached its peak value (476.576 + 317.067ng/ml; 481.177 + 322.763ng/ml). The difference was statistically significant (t=3.18, P=0.025; t=3.159, P=0.025) compared with before embolization (t=3.18, P=0.025; t=3.159, P=0.025). To the 2-3 day, the difference was still high (382.956 +, 381.943 + 346.108ng/ml), but with embolism. The difference was not statistically significant (t=2.361, P=0.065; t=2.327, P=0.067).
Group Sham: after D-dimer level, 3hs increased (450.881 + 165.966ng/ml) and 24hs reached a peak (504.488 + 124.615ng/ml). Compared with preoperative (51.050 + 6.795ng/ml), the difference was significant (t=6.071, P=0.000; t=9.170, P=0.000).2-3 days resumed normal levels.
conclusion
1. elevated coagulation fibrinolytic markers, plasma SFC, F III and D-dimer levels, are early sensitive indicators of experimental VTE, and are dynamic, but not specific.
2.DVT and PTE are different stages of VTE, and the expression of coagulation and fibrinolytic markers is not the same. DVT is a dynamic process of thrombus formation and simultaneous dissolution of SFC under the cross linking of F III. While PTE has been dissolved mainly after embolic embolism, although SFC and F III have a higher trend, but there is no significant difference from before embolism.
3. surgical trauma can lead to elevated coagulation and fibrinolytic markers, plasma levels of SFC, F, III and D-dimer, and is a predisposing factor for VTE.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.5

【参考文献】