中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
10期
905-908
,共4页
杜春蔚%王秀娟%赵丽丽%戴克胜
杜春蔚%王秀娟%趙麗麗%戴剋勝
두춘위%왕수연%조려려%대극성
他克莫司%流式细胞术%血小板聚集%细胞凋亡
他剋莫司%流式細胞術%血小闆聚集%細胞凋亡
타극막사%류식세포술%혈소판취집%세포조망
Tacrolimus%Flow cytometry%Platelet aggregation%Apoptosis
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.
目的 觀察他剋莫司在體外對血小闆功能的影響.方法 取18～25歲、2週內未服用抗血小闆藥物的健康誌願者靜脈血,製備洗滌血小闆,分彆加入0.06、0.6、6、60、120、240 μmol/L他剋莫司,37℃孵育2h,用流式細胞術檢測血小闆線粒體跨膜電位和P-選擇素變化,用Western blot方法檢測血小闆凋亡蛋白的錶達,用血小闆聚集儀檢測血小闆聚集功能的改變.結果 0.06 μmol/L他剋莫司可以促進膠原蛋白誘導的血小闆聚集、抑製凝血酶誘導的血小闆聚集,對瑞斯託黴素和血管性血友病因子誘導的血小闆聚集沒有影響.120、240 μmol/L他剋莫司引起血小闆線粒體膜電位降低和半胱氨痠天鼕氨痠蛋白酶3(caspase-3)凋亡蛋白的錶達.結論 高濃度的他剋莫司可引起血小闆凋亡;0.06μmol/L(相噹于50 ng/ml血藥濃度)的他剋莫司按刺激劑不同對血小闆聚集功能的影響不同.
목적 관찰타극막사재체외대혈소판공능적영향.방법 취18～25세、2주내미복용항혈소판약물적건강지원자정맥혈,제비세조혈소판,분별가입0.06、0.6、6、60、120、240 μmol/L타극막사,37℃부육2h,용류식세포술검측혈소판선립체과막전위화P-선택소변화,용Western blot방법검측혈소판조망단백적표체,용혈소판취집의검측혈소판취집공능적개변.결과 0.06 μmol/L타극막사가이촉진효원단백유도적혈소판취집、억제응혈매유도적혈소판취집,대서사탁매소화혈관성혈우병인자유도적혈소판취집몰유영향.120、240 μmol/L타극막사인기혈소판선립체막전위강저화반광안산천동안산단백매3(caspase-3)조망단백적표체.결론 고농도적타극막사가인기혈소판조망;0.06μmol/L(상당우50 ng/ml혈약농도)적타극막사안자격제불동대혈소판취집공능적영향불동.
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 (0.06,0.6,6,60,120,240 μmol/L) at 37 ℃ for 2 hours,and then the changes of mitochondrial membrane potential and P-selectin 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 function.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 0.06 μmol/L (which is equivalent to 50 ng/ml blood concentration) could have different effects on platelet aggregation function according to different stimulating agents.
