摘要
目的 测定磷酯酶C(phospholipaseC ,简称PLC ,下同 )对静息状态和激活状态下人血小板胞质游离钙离子浓度的影响 ,从而进一步阐明PLC抗血小板聚集作用的机制。方法 将健康成人的洗涤血小板用荧光指示剂Fura 2 /AM进行负载 ,分别用生理盐水、ASA和不同剂量的PLC处理荧光负载后的血小板 ,采用双波长荧光分光光度法分别测定经过不同处理后的血小板在静息状态和以ADP为诱导剂时的激活状态下的胞质游离钙离子浓度 [Ca2 + ]i。结果 以健康成人血为样品时 ,经生理盐水、ASA 334μmol·L-1、2 5、3 75、5、10和 2 0UPLC·ml-1处理后的血小板 ,在静息状态下测得的胞质游离 [Ca2 + ]i 浓度 (nmol·L-1)分别为15 2 5 5± 15 0 7,131 6 3± 15 5 8,14 0 2 7± 12 0 3,139 4 8± 1 73,12 1 11± 9 5 8,116 6 2± 15 96和 10 7 2 0± 17 0 7,而以ADP为诱导剂激活血小板后测得的胞质游离 [Ca2 + ]i(nmol·L-1)分别为 90 2 6 2± 94 74 ,6 87 99± 6 2 86 ,810 99± 72 37,70 1 73± 2 1 37,4 2 9 6 7± 71 5 9,342 82± 4 4 86和 2 6 3 2 7± 2 5 4 6。以生理盐水作为对照 ,ASA 334μmol·L-1、2 5、3 75、5、10和 2 0UPLC·ml-1剂量组对激活状态下血小板胞质 [Ca2 + ]i 的抑制率I(% )分别为 2 5
AIM Aim to study the effect of PLC on human platelet cytoplasmic free calcium concentration, and then to explore the mechanism of PLC anti aggregation to platelet. METHODS Washed platelet of human was marked with fluorescence indicator Furu 2/AM, and then was treated with PSS, ASA and different dose of PLC respectively. Cytoplasmic free [Ca 2+ ] i of differently treated platelet at static state and activated sate induced by ADP was determined by double beam fluorescence spectrophotometric method. RESULTS Healthy human platelet was treated with PSS, ASA 334 μmol·L -1 , 2 5, 3 75, 5, 10 and 20U PLC·ml -1 . Cytoplasmic [Ca 2+ ] i determined at static state was 152 55±15 07, 131 63±15 58, 140 27±12 03, 139 48±1 73, 121 11±9 58, 116 62±15 96 and 107 20±17 07 respectively. [Ca 2+ ] i determined at activated sate induced by ADP was 902 62±94 74, 687 99±62 86, 810 99±72 37, 701 73±21 37, 429 67±71 59, 342 82±44 86 and 263 27±25 46 respectively. Compared with PSS, inhibition rates ( % ) of ASA 334 μmol·L -1 , 2 5, 3 75, 5,10, 20 U PLC·ml -1 to the increment of activated platelet cytoplasmic [Ca 2+ ] i were 25 83, 10 58, 25 04, 58 86, 69 84, 79 19 respectively. At the same condition, inhibition rate ( % ) of 10 U PLC·ml -1 to the increment of five hypertensive activated platelet cytoplasmic [Ca 2+ ] i were 74 25, 73 48, 61 32, 82 87,70 60 respectively. CONCLUSION PLC has significant effects on static cytoplasmic[Ca 2+ ] i of healthy and hypertensive human platelet ( P <0 05), and has significant dose dependant inhibition on increment of activated cytoplasmic [Ca 2+ ] i ( P <0 05). All these indicate PLC can inhibit internal calcium discharge.This may be one of the important reason why PLC can inhibit platelet aggregation.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2004年第1期37-40,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助课题 No 3 0 1710 74
国家"十五"攻关课题
No 2 0 0 1BA 707B01
中国科学院创新工程重大项目(kscx 1 sw 11)资助项目
关键词
磷酯酶C
人血小板
胞质游离钙
荧光测定
phospholipase C
human platelet
cytoplasmic free calcium
fluorescence measurement
