他克莫司对血小板功能影响的体外研究  

In vitro effects of tacrolimus on platelet function

作　　者：杜春蔚[1] 王秀娟[1] 赵丽丽[1] 戴克胜[1] 

机构地区：[1]苏州大学附属第一医院、江苏省血液研究所,卫生部血栓与止血重点实验室,血液学协同创新中心,215006

出　　处：《中华血液学杂志》2014年第10期905-908,共4页Chinese Journal of Hematology

基　　金：科技部973前期研究专项（2012CB526609）;国家自然科学基金重点项目（81130008）;江苏省科教兴卫工程-临床医学中心资助（ZX201102）

摘　　要：目的 观察他克莫司在体外对血小板功能的影响.方法 取18～25岁、2周内未服用抗血小板药物的健康志愿者静脉血,制备洗涤血小板,分别加入0.06、0.6、6、60、120、240 μmol/L他克莫司,37℃孵育2h,用流式细胞术检测血小板线粒体跨膜电位和P-选择素变化,用Western blot方法检测血小板凋亡蛋白的表达,用血小板聚集仪检测血小板聚集功能的改变.结果 0.06 μmol/L他克莫司可以促进胶原蛋白诱导的血小板聚集、抑制凝血酶诱导的血小板聚集,对瑞斯托霉素和血管性血友病因子诱导的血小板聚集没有影响.120、240 μmol/L他克莫司引起血小板线粒体膜电位降低和半胱氨酸天冬氨酸蛋白酶3（caspase-3）凋亡蛋白的表达.结论 高浓度的他克莫司可引起血小板凋亡;0.06μmol/L（相当于50 ng/ml血药浓度）的他克莫司按刺激剂不同对血小板聚集功能的影响不同.Objective To investigate the in vitro effects of immune inhibitor tacrolimus on platelet function. Methods Fresh venous blood was collected from healthy volunteers at ages of 18-25 years old, who are not taking antiplatelet drugs within two weeks. The platelets were isolated from the blood and incubated with different concentrations of tacrolimus （0.06, 0.6, 6, 60; 120, 240μmol/L） at 37℃ for 2 hours, and then the changes of mitochondrial membrane potential and Pselectin of platelets were detected by flow cytometry, the expression of apoptosis related protein by Western Blot, and the change of the platelet aggregation function by platelet aggregation analyzer. Results Tacrolimus at concentration of 0.06 μmol/L could promote collagen induced platelet aggregation, inhibit thrombin induced platelet aggregation, have no effect on ristocetin and vWF induced platelet aggregation ffmction. Tacrolimus at concentration of 120 μmol/L and 240 μmol/L could reduce the platelet mitochondrial membrane potential and induce the expression of apoptosis protein caspase-3. Conclusion In vitro experimental results showed that high concentration of tacrolimus could lead to platelet apoptosis. But the current therapeutic dose of tacrolimus at 0.06μmol/L （which is equivalent to 50 ng/ml blood concentration） could have different effects on platelet aggregation function according to different stimulating agents.

关 键 词：他克莫司 流式细胞术 血小板聚集 细胞凋亡 

分 类 号：R558[医药卫生—血液循环系统疾病]