论文部分内容阅读
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?
Objective To determine the effect of phospholipase C (PLC) on the ADP-induced aggregation of rabbit and human platelet actin in order to elucidate the mechanism of PLC antiplatelet aggregation. Methods The PRP of rabbits and healthy adults were treated with normal saline, ASA and different doses of PLC respectively. ADP was used to induce activation. Actin was extracted by extraction with Triton extract and SDS PAGE, and then determined by spectrophotometry Actin relative content. Results The relative content of actin in rabbit platelets treated with saline was 1 6 82 ± 0 319. The relative content of actin in the platelets activated by ADP was measured by saline, ASA 6 6 8μmol·L-1, PLC 5,10,15,2 0 and 25 U · ml-1 Were 2450 ± 0 5 6 2, 1 0 89 ± 0 32 2, 1 727 ± 0 4 4 2, 1 45 ± 0 32 4, 1 16 1 ± 0 30 6, 0 85 7 ± 0 2 4 2 and 0 6 92 ± 0 187. Compared with saline group, the inhibition rates of platelet actin polymerization in all treatment groups were 555, 295, 1482, 526, 16502 and 7176, respectively . The relative content of actin in human platelets treated with saline was 1 35 8 ± 0 376. The relative content of actin in platelets after treatment with physiological saline, ASA 6 6 8μmol·L-1, PLC 5,10,15,2 0 and 25 U · ml-1 and activated by ADP respectively Were 2444 ± 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 ?