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目的探讨正常人血小板是否含有组织因子(TF),血小板相关TF是否具有促凝活性。方法用Sepharose2B凝胶柱分离纯化血小板,ELISA法测定洗涤血小板破碎液TF含量;一期血浆凝固时间测定血小板相关TF促凝活性;RT-PCR法检测正常人血小板相关TF基因的表达。结果正常人血小板破碎液中TF含量为(16.37±6.39)ng/L;胶原活化的血小板可将TF释放到血浆中引起血浆TF水平明显升高(P<0.05);静息血小板无TF的促凝活性,胶原活化的血小板促凝活性显著增高(P<0.01),TF单抗及乏凝血因子Ⅶ(FⅦ)血浆能明显阻断该效应(P<0.01);RT-PCR法在血小板上未能检测到TF mRNA的存在;冠心病患者血小板破碎液TF水平为(20.71±8.78)ng/L,与正常对照相比明显增高(P<0.05),体外未经胶原活化的血小板促凝活性也较正常对照明显增高,并能被TF单抗抑制。结论血小板内含有TF;活化血小板可表现出TF促凝活性;血小板所含的TF可能并非自身合成;冠心病患者血小板相关TF的含量及活性较正常对照明显增高,提示血小板可能可以通过释放TF而参与凝血过程及血栓形成。
Objective To investigate whether normal human platelets contain tissue factor (TF) and whether platelet-associated TF has procoagulant activity. Methods Sepharose 2B column was used to separate and purify the platelets. ELISA was used to determine the content of TF in washing platelets. The plasma coagulation time was used to measure the procoagulant activity of platelet-associated TF. The expression of TF gene in platelets was detected by RT-PCR. Results The content of TF in the normal human platelet crush fluid was (16.37 ± 6.39) ng / L. Collagen-activated platelets could release TF into the plasma and significantly increased the level of TF in plasma (P <0.05). (P <0.01). The effect of TF monoclonal antibody and plasma of coagulation factor Ⅶ (F Ⅶ) on blood coagulation activity and collagen activated platelet aggregation was significantly inhibited (P <0.01) The level of TF mRNA was detected in the patients with coronary heart disease (20.71 ± 8.78ng ng / L), which was significantly higher than that of the normal controls (P <0.05). In addition, the activity of platelet procoagulant without collagen activation Compared with the normal control, it was significantly increased and inhibited by TF monoclonal antibody. Conclusion Platelet contains TF; activated platelets can exhibit TF procoagulant activity; platelet contains TF may not be their own synthesis; coronary heart disease patients with platelet-associated TF content and activity was significantly higher than the normal control, suggesting that platelets may be released by TF Participate in coagulation process and thrombosis.