雷洛昔芬与结合雌激素联合应用对去势大鼠血栓前状态的影响

Effects of raloxifene combined with conjugated equine estrogen on prothrombotic state in ovariectomized rats

目的通过去势建立绝经大鼠模型，将雷洛昔芬（RLX）与不同剂量的结合雌激素（CEE）配伍，观察两药联合应用对绝经大鼠血栓前状态的影响。方法将64只9～10月龄的健康雌性SD大鼠随机分为4组，每组8只。手术去势后1周进行药物干预，给药10周后处死。对照组：给予安慰剂（0．9％氯化钠溶液灌胃）；RLX组：给予RLX6mg／（kg·d）；CEE组：给予CEE0．07mg／（kg·d）；RLX＋低量CEE组：给予RLX6mg／（kg·d）＋CEE0．07mg／（kg·d）；RLX＋高量CEE组：给予RLX6mg／（kg·d）＋CEE0．5mg／（kg·d）。采用凝血酶比浊法检测各组大鼠血浆纤维蛋白原（FIB）含量；采用发射底物法检测抗凝血酶Ⅲ（ATⅢ）活性；采用ELISA双抗体夹心法检测纤溶酶原激活物抑制剂1（PAI-1）、D二聚体（D—D）、血管性假血友病因子（vwF）含量；采用硝酸还原酶法检测NO含量。结果（1）凝血及抗凝血指标：对照组FIB含量为（1．62±0．22）g／L、RLX组为（2．02±0．54）g／L、RLX＋低量CEE组为（1．97±0．16）g／L、RLX＋高量CEE组为（2．00±0．18）g／L，后3组分别与对照组比较，差异均有统计学意义（P〈0．05）；但后3组间比较，差异无统计学意义（P〉0．05）。血浆ATⅢ活性各组间比较，差异均无统计学意义（P〉0．05）。（2）纤溶和抗纤溶指标：对照组PAI-1含量为（14．1±2．8）μg／L，RLX组为（20．0±3．3）μg／L，RLX＋低量CEE组为（41．5±5．5）μg/L，RLX＋高量CEE组为（38．9±6．0）μg/L，后3组分别与对照组比较，差异均有统计学意义（P〈0．05）；但血浆D—D含量各组间比较，差异无统计学意义（P〉0．05）。（3）血管内皮功能：对照组血浆vwF含量为（43±7）％，RLX组为（49±5）％，RLX＋低量CEE组为（46±6）％，RLX＋高量CEE组为（36±5）％，RLX＋高量CEE组血浆vwF含量明显低于其他3组，差异也有统计学意义（P〈0．05）。各组NO含量比较，差异无统计学意义（P〉0．05）。结论在血栓形成的不同环节，RLX发挥了不同的效应，可能导致血栓形成的风险增加。CEE在纤溶和抗纤溶方面与RLX具有协同作用，可进一步增加RLX的致血栓效应；在血管内皮功能方面则起到一定保护作用，然而这种效应需要高剂量CEE才能显现。

Objective To study effects of raloxifene （RLX） with different doses of conjugated equine estrogen（CEE） on prothrombotic profiles in the ovariectomized rats model. Methods Total of 32 healthy female SD rats at age of 9 to 10 months were equally divided into every 8 rats at 4 groups randomly. One week after ovariectomized, they were treated by drugs, including control group with placebo（0. 9% Nael intragastrie administration） , RLX group with RLX 6 rag/（ kg· d） , RLX and low CEE group with RLX 6 mg/（ kg · d） ＋ CEE 0. 07 mg/（ kg · d） and RLX and high CEE group with RLX 6 rag/（ kg · d） ＋ CEE 0. 5 mg/（kg·d）for 10 weeks before death. Thrombin turbidimetry method was used to evaluate the plasma fibrinogen（ FIB ）, transmitting substrate method for antithrombin Ⅲ （ AT Ⅲ） activity, double-antibody sandwich ELISA for plasminogen activator inhibitor 1 （PAI-1）, D-dimer （D-D） and yon Willebrand factor （vWF） and nitrate reductase method for nitric oxide （NO）. Results （ 1 ） Coagulation and anticoagulation indieators ： it was observed （ 1.62 ± 0. 22） g/L FIB at control group, （2. 02 ± 0. 54） g/L at RLX group, （1.97 ±0. 16） g/L at RLX and low CEE group, （2.00 ±0. 18） g/L at RLX and high CEE group. There was a statistically significant difference between control group and any one of treatment groups（ P 〈 0. 05 ）and no statistical significance among those three treatment groups （ P 〉 0. 05 ）. No significant change was observed in plasma AT Ill activity among groups （ P 〉 0.05 ）. （ 2 ） Fibrinolytic and anti-fibrinolytic indicators ： it was observed （14. 1 ±2. 8） μg/L PAI-1 at control group, （20. 0±3.3） μg/L at RLX group, （41.5± 5.5） μg/L at RLX and low CEE group, （38.9 ±6.0） μg/L at RLX and high CEE group. A remarkable increase was observed between control group and any one of treatment groups（P 〈0. 05）. But there was no significant difference of D-D among groups（P 〉0. 05）. （3） Endothelial function： it was （43±7）% vWF at control group, （49±5）% at RLX group, （46 ±6）% at RLX and low CEE group, （36±5）% at RLX and high CEE group. The vWF of RLX and high CEE group was the lowest among all groups （P 〈 0. 05 ）. There was no differenee of NO among groups （ P 〉 0. 05 ）. Conclusions In the different links of thrombosis, RLX gives different fuetion and may increase the risk. CEE plays a synergism role in the matter of fibrinolysis and anti-fibrinolysis with RLX, further giving rise to thrombosis effect of RLX. And it also has a protective role in the function of vascular endothelium to some extent. But this only works with high dose.