摘要
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 explore whether normal platelet contains tissue factor （TF） , and the significance of platelet-associated TF（PATF）. Methods Platelets were isolated by Sepharose 2B gel colunm. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction（RT-PCR）. Results A certain amount of TF antigen（ 16.37 ± 6.39）ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma（ P 〈 0.05 ）. Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor Ⅶ plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls （P 〈0.05 ）. Resting platelets from the patients showed a higher procoagulant activity, which could he inhibited by TF McAb. Conclusion Platelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may he involved in coagulation and thrombosis by releasing TF.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2005年第9期525-528,共4页
Chinese Journal of Hematology
