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目的:观察家兔口服纳豆激酶(NK)对血液凝血和纤溶功能的影响。方法:32只新西兰白兔随机分为4组,每组8只,包括空白对照组(A组)、模型组(B组)、低剂量NK组(C组)和高剂量NK组(D组),常规高脂饮食构建动脉粥样硬化模型。采用凝固法测定血浆凝血酶原(PT)、部分凝血活酶时间(APTT)和凝血因子Ⅰ(Fg)浓度;ELISA法测定血液纤溶酶原激活剂(tPA)和纤溶酶原激活剂抑制物(PAI-1)浓度、RT-PCR法检测主动脉组织tPA和PAI-1 mRNA的表达、免疫组化法检测主动脉组织PAI-1蛋白表达。结果:与A组相比,B组的PT明显缩短,C、D组的APTT明显延长,Fg含量明显下降(P<0.01),以D组更明显;D组的tPA浓度明显增加[(0.91±0.17)∶(1.25±0.23)μg/L,P<0.05],B组的PAI-1显著增加[(2.00±0.45)∶(2.75±0.84)μg/L,P<0.05];C、D组的tPA mRNA表达较B组明显增加(分别为1.22±0.23和1.33±0.16∶0.64±0.10,P<0.01),PAI-1 mRNA则显著减少(分别为1.34±0.17和1.31±0.09∶1.90±0.18,P<0.01);免疫组化染色显示A组、B组、C组和D组的单位面积动脉内膜及中膜PAI-1阳性染色标记物的平均A值分别为(155.86±28.19)、(190.49±19.80)、(148.52±9.86)和(138.80±3.13),P<0.05或P<0.01。结论:NK可明显提高血液纤溶活性、降低凝血活性,显著增加动脉硬化组织tPA mRNA的表达、减少PAI-1 mRNA和蛋白的表达。
Objective: To observe the effects of natto kinase (NK) on blood coagulation and fibrinolysis in rabbits. Methods: Thirty-two New Zealand white rabbits were randomly divided into 4 groups with 8 rats in each group, including blank control group (A group), model group (B group), low dose NK group (C group) and high dose NK group ), Conventional high-fat diet to construct atherosclerosis model. The concentrations of plasma prothrombin (PT), partial thromboplastin time (APTT) and coagulation factor Ⅰ (Fg) were measured by coagulation method. The levels of plasminogen activator (tPA) and plasminogen activator (PAI-1), the expression of tPA and PAI-1 mRNA in aortic tissue were detected by RT-PCR, and the expression of PAI-1 in aortic tissue was detected by immunohistochemistry. Results: Compared with group A, PT in group B was significantly shortened, APTT in group C and group D were significantly prolonged, and Fg content was significantly decreased (P <0.01), especially in group D; the level of tPA in group D was significantly increased (2.00 ± 0.45) :( 2.75 ± 0.84) μg / L, P <0.05]; C, D (± 0.17): (1.25 ± 0.23) μg / L, (1.22 ± 0.23 and 1.33 ± 0.16:0.64 ± 0.10, respectively, P <0.01), while PAI-1 mRNA was significantly decreased (1.34 ± 0.17 and 1.31 ± 0.09:1.90 ± respectively 0.18, P <0.01). The average A values of PAI-1 positive staining markers per unit area of arterial intima and media in group A, group B, group C and group D were (155.86 ± 28.19) , (190.49 ± 19.80), (148.52 ± 9.86) and (138.80 ± 3.13) respectively, P <0.05 or P <0.01. Conclusion: NK can significantly increase blood fibrinolytic activity, reduce coagulation activity, significantly increase atherosclerosis tissue tPA mRNA expression, reduce PAI-1 mRNA and protein expression.