鸦胆子油对血小板聚集的抑制作用及其机制的体外研究  

The in vitro inhibitory effects of brucea javanica oil on platelet aggregation and the related mechanism

作　　者：费鲜明[1] 潘建平[2] 吴建国[1] 蒋雷[1] 王珍妮[1] 周永列[1] 

机构地区：[1]浙江省人民医院检验医学中心,杭州310014 [2]浙江大学免疫学研究所

出　　处：《中华检验医学杂志》2010年第1期68-72,共5页Chinese Journal of Laboratory Medicine

基　　金：浙江省医学重点学科基金（07-010）

摘　　要：目的以人洗涤血小板为模型，体外观察鸦胆子油对血小板聚集的抑制作用，并探讨其可能机制。方法将不同浓度鸦胆子油与人洗涤血小板作用，分为4组：空白组（加丙酮）、阴性对照组（加丙酮，0％），9．0％鸦胆子油组及22．5％鸦胆子油组。以血小板聚集分析仪检测ADP、AA、胶原和凝血酶诱导的血小板最大聚集率、以流式细胞仪检测活化血小板膜FIB．R和P-选择素表达水平，以SDS—PAGE分析血小板肌动蛋白聚合体的变化。结果9．0％鸦胆子油组ADP、AA、胶原和凝血酶诱导的血小板聚集率分别为（57．7±4．0）％、（62．2±3．9）％、（66．9±5．0）％和（71．8±5．1）％，均低于阴性对照（0％）组的（75．3±4．1）％、（79．3±4．8）％、（80．6±5．4）％和（84．1±6．2）％，差异均有统计学意义（t值分别为7．312、8．785、9．123和9．466，P均〈0．01）；22．5％组分别为（39．2±3．5）％、（45．8±3．4）％、（51．2±3．9）％和（56．7±4．8）％，显著低于9．0％组，差异有统计学意义（t值分别为9．123、10．221、9．533和10．243，P均〈0．01）。22．5％鸦胆子油组对血小板聚集的抑制率分别为47．9％、42．2％、36．5％和32．6％，显著高于9．0％组的23．4％、21．6％、17．0％和14．6％，差异有统计学意义（Х^2值分别为13．08、9．77、9．70和8．93，P均〈0．01）。血小板聚集水平与鸦胆子油浓度呈负相关（r=-0．952，-0．961，-0．970和-0．975，P〈0．01）。9．0％鸦胆子油组ADP诱导的血小板FIB—R表达水平和P-选择素表达水平分别为（64．7±4．0）％、（3．91±0．21）％，均低于阴性对照组的（85．5±4．6）％和（5．05±0．27）％，差异有统计学意义（t值分别为11．35、7．81，P均〈0．01）；22．5％鸦胆子油组分别为（36．2±3．9）％和（2．34±0．15）％，低于9．0％�Objective To observe the in vitro inhibitory effects of brucea javanica oil on human washed platelet and explore the possible mechanism. Methods Human washed platelets were mixed with different concentration of brucea javanica oil which were divided into four groups [ untreated control group, negative control group,9. 0% of brucea javanica oil group, and 22.5% of brucea javanica oil group]. The maximal ratio of platelet aggregation induced by adenosine liphosphate （ ADP）, arachidonic acid （ AA）, collagen, and thrombin, respectively, was measured with platelet aggregation analyzer. The expressions of flbrinogen receptor （FIB-R） and p-selectin （CD62p ） on external membrane of activated platelet were determinated with flow cytometry. The F-actin of cytoskeletal structure in activated platelet was detected by SDS-PAGE. Results At 9. 0% of brucea javanica oil, the maximal ratio of platelet aggregation [ （ 57.7 ± 4.0）%, （62.2 ±3.9）%, （66.9 ±5.0）% and （71.8 ±5.1）% ] induced by ADP, AA, collagen, and thrombin, was significantly lower than that [ （75.3 ± 4. 1 ） % , （79.3 ± 4. 8 ） %, （ 80. 6 ± 5.4 ） % , （ 84. 1 ± 6. 2） % ] at negative control （0% of brueea javanica oil ） （ P 〈 0. 01 ）, but makedly higher than that [ （ 39. 2 ± 3.5 ） %, （ 45.8 ± 3.4 ） %, （ 51.2 ± 3.9 ） % and （ 56. 7 ± 4. 8 % ） ] at 22. 5 %, respectively （ P 〈 0. 01 ）. The inhibitory rate of platelet aggregation （ 47.9% , 42. 2% , 36. 5% and 32.6% ） at 22. 5% of brueea javaniea oil was notably higher than that （ 23.4% ,21.6% , 17.0% and 14. 6% ） at 9. 0% , respectively （ P 〈 0. 001 ）. There was a negative correlation between brucea javanica oil concentration and the aggregation ratio of platelet stimulated by the four agonists, respeetively （ r = - 0. 952, - 0. 961, - 0. 970, - 0. 975, P 〈 0. 001 ）. At 9. 0% of brueea javanica oil, the expression levels of FIB-R [ （ 64. 7 ± 4. 0 ） % ] and CD62p [ （3.91 ±0. 21）% ] of platelet activated by A

关 键 词：鸦胆子素 血小板聚集 受体 纤维蛋白原 P选择素 肌动蛋白类 

分 类 号：R285[医药卫生—中药学]