磷脂酶C对ADP诱导的兔、人血小板肌动蛋白聚合的抑制作用

目的　测定磷脂酶C(PLC)对ADP诱导的兔、人血小板肌动蛋白聚合的的影响 ,从而进一步阐明PLC抗血小板聚集作用的机制。方法　取家兔和健康成人的PRP分别用生理盐水、ASA和不同剂量的PLC处理 ,以ADP诱导激活 ,采用Triton抽提液提取和SDS PAGE分离出肌动蛋白 ,再以分光光度法分别测定其肌动蛋白相对含量。结果　家兔血小板经生理盐水处理后的肌动蛋白相对含量为 1 6 82±0 319。而经生理盐水、ASA 6 6 8μmol·L-1、PLC 5、10、15、2 0和 2 5U·ml-1各组处理 ,并以ADP激活的血小板 ,测得的肌动蛋白相对含量分别为 2 4 5 0± 0 5 6 2 ,1 0 89± 0 32 2 ,1 72 7± 0 4 4 2 ,1 4 5 0± 0 32 4 ,1 16 1± 0 30 6 ,0 85 7± 0 2 4 2和0 6 92± 0 187。以生理盐水组作为对照 ,各处理组血小板肌动蛋白聚合的抑制率I(% )分别为 5 5 5 5 ,2 9 5 1,4 0 82 ,5 2 6 1,6 5 0 2 ,71 76。人血小板经生理盐水处理后的肌动蛋白相对含量为 1 35 8± 0 376。而经生理盐水、ASA 6 6 8μmol·L-1、PLC 5、10、15、2 0和 2 5U·ml-1各组处理并以ADP激活后的血小板 ,测得的肌动蛋白相对含量分别为2 4 4 5± 0 75 0 ,1 0 96± 0 344 ,1 70 5± 0 5 0 7,1 4 37±0 4 16 ,1 16 5± 0 35 5 ,0 84 5± 0 2 5 7?

Effect of PLC on rabbit and human platelet actin polymerization induced by ADP

Aim To study the effect of PLC on rabbit and human platelet actin polymerization, and then to explore the mechanism of PLC anti-aggregation to platelet. Methods Platelets of rabbit and human were treated with PSS, ASA and different doses of PLC respectively and then were extracted by Triton abstraction. The relative concentration of actin of differently treated platelets induced by ADP was determined by SDS-PAGE and spectrophotometre. Results For rabbit platelets were treated with PSS, the relative concentration of actin determined at static state was 1.682±0.319; when the platelets were treated with ASA 668 μmol·L -1,PLC 5,10,15,20 and 25 U·ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.450±0.562,1.089±0.322,1.727±0.442,1.450±0.324,1.161±0.306, 0.857±0.242 and 0.692±0.187 respectively. Compared with PSS, inhibition rates (%) of ASA 668 μmol·L -1, PLC 5, 10, 15, 20, 25 U·ml -1 to the relative concentration of actin were 55.55,29.51, 40.82,52.61, 65.02,71.76 respectively.For human platelets were treated with PSS, the relative concentration of actin determined at static state was 1.358±0.376; when the platelets were treated with ASA 668 μmol·L -1,PLC 5,10,15,20 and 25 U·ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.445±0.750, 1.096±0.344, 1.705±0.507,1.437±0.416, 1.165±0.355, 0.845±0.257 and 0.679±0.198 respectively. Compared with PSS, inhibition rates (%) of ASA 668 μmol·L -1, PLC 5, 10, 15, 20, 25 U·ml -1 to the relative concentration of actin were 55.17,30.27, 41.23,52.35, 65.44, 72.23 respectively. Conclusion PLC has significant effects on actin polymerization of rabbit and healthy human platelets (P<0.05), and has significant dose-dependant inhibition on actin polymerization (P<0.05). All these indicate PLC can inhibit actin polymerization. This may be one of the important reasons why PLC can inhibit platelet aggregation.