磷脂酶C对ADP诱导的兔血小板IP_3水平的影响

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目的测定磷酯酶C(PLC)对ADP诱导的兔血小板三磷酸肌醇(IP3)的影响,从而进一步阐明PLC抗血小板聚集作用的机制.方法取家兔血除去红细胞制备血小板,血小板计数后分为8个组.第1组用生理盐水处理,第2组用ADP处理,第3组用ASP处理以ADP诱导激活,第4~8组分别用不同剂量的PLC处理并以ADP诱导激活.采用三氯醋酸法制备血小板IP3,再用放射免疫分析法分别测定IP3含量.结果家兔血小板经生理盐水处理后的IP3含量为0.29 pmol/108血小板,而经ADP处理后IP3含量为0.39pmol/108血小板.经ASP 668μmol.L-1、5、10、15、20和25 U PLC.ml-1各组处理,并以ADP激活的血小板,测得的IP3含量分别为0.19、0.08、0.15、0.25、0.04和0.18(pmol/108血小板).ADP组比生理盐水组IP3有明显的提高,而ASP组、不同的PLC组比ADP组IP3有明显的降低(P“,”Aim To determine the effect of phospholipase C(PLC) on the rabbit platelet triphosphoryl inositol(IP3) induced by ADP,and then to explore the mechanism of anti-aggregation to platelet by PLC.Method PRP was made from rabbit blood by removing red cells,and then was parted to eight groups.The first group was treated with PSS.The second was treated with with ADP.The third was treated with ASP and then induced by ADP.Four to eight groups were treated with different dose of PLC respectively and then induced by ADP.IP3 of rabbit platelet was made by trichloro-acetic acid method,and its content was tested by radioimmunological assay.Result IP3 of rabbit platelet treated with PSS was 0.29 pmol/108 platelet.IP3 of rabbit platelet treated with ADP was 0.39 pmol/108 platelet.IP3 of rabbit platelet,which were treated with ASP 668 μmol·L-1,5,10,15,20 and 25 U PLC·ml-1 respectively and then induced by ADP,were 0.29,0.19,0.08,0.15,0.25,0.04 and 0.18 pmoL/108 platelet respectively.IP3 of ADP group was apparently higher than that of PSS group,but the IP3 of ASP group and different dose PLC group were significantly lower than ADP group(P<0.05).Conclusion PLC can decrease the IP3 of rabbit platelet induced by ADP,and this effect does not indicate the relationship between dose and effect.The study testifies PLC has function of decreasing IP3 of rabbit platelet.This may be one of the important reasons of anti-aggregation to platelet of PLC.
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