血浆凝块收缩功能实验方法学研究  

Study on the Experimental Methodology of Plasma Clot Retraction

作　　者：罗扬淦 卢姿含 廖晗婧 郝逗逗 黄镘静 朱枝祥 LUO Yang-Gan;LU Zi-Han;LIAO Han-Jing;HAO Dou-Dou;HUANG Man-Jing;ZHU Zhi-Xiang(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院,北京100029 [2]北京中医药大学北京中医药研究院,北京100029

出　　处：《中国实验血液学杂志》2024年第4期1271-1277,共7页Journal of Experimental Hematology

摘　　要：目的:探讨影响血浆凝块收缩功能的关键因素,优化血浆凝块收缩功能实验方法,以用于研究血小板功能调控和评价药物对血浆凝块收缩功能的调节作用。方法:依次研究不同浓度的凝血酶、Ca^(2+)以及血小板对血浆凝块收缩率的影响,优化血浆凝块收缩检测体系。通过检测多种血小板信号通路抑制剂对血浆凝块收缩的调节作用验证该检测体系的有效性。结果:通过对凝血酶浓度、Ca^(2+)浓度和血小板浓度的优化研究,利用0.2 U/ml凝血酶处理含0.5 mmol/L Ca^(2+)以及40×10^(9)/L血小板的富血小板血浆(PRP)进行血浆凝块收缩分析。在此条件下,凝血酶处理15 min能够显著诱导血浆凝块收缩反应,处理30 min诱导的血浆凝块收缩率大于50%。利用此检测条件研究抑制剂对血浆凝块收缩的调节作用,PKC抑制剂Go 6983能够强烈抑制血浆凝块收缩反应,PI3K抑制剂Ly294002和p38 MAPK抑制剂SB203580能够轻度抑制血浆凝块收缩反应。结论:利用0.2 U/ml凝血酶处理含0.5 mmol/L Ca^(2+)和40×10^(9)/L血小板的PRP进行血浆凝块收缩检测,可以用于研究血小板功能调控和评价药物对血浆凝块收缩的调节作用。Objective:To explore the key factors affecting plasma clot retraction and optimize the experimental method of plasma clot retraction,in order to study the regulation of platelet function and evaluate the modulatory effects of drugs on plasma clot retraction.Methods:The effects of different concentrations of thrombin,Ca^(2+)and platelets on plasma clot retraction were studied,and the detection system of plasma clot retraction was optimized.The availability of the detection system was then validated by analyzing the regulatory effects of multiple signaling pathway inhibitors on plasma clot retraction.Results:Through the optimization study of multiple factors,platelet rich plasma(PRP)containing 0.5 mmol/L Ca^(2+)and 40×10^(9)/L platelets was treated with 0.2 U/ml thrombin to perform plasma clot retraction analysis.After treatment with thrombin for 15 min,plasma clot retracted significantly.After treatment with thrombin for 30 min,the percentage of plasma clot retraction was more than 50%.The regulatory effects of multiple signaling pathway inhibitors on plasma clot retraction were studied in this detection system.PKC inhibitor Go 6983 exhibited a significant inhibitory effect on plasma clot retraction,while PI3K inhibitor Ly294002 and p38 MAPK inhibitor SB203580 slightly suppressed plasma clot retraction.Conclusion:PRP containing 0.5 mmol/L Ca^(2+)and 40×10^(9)/L platelets can be induced with 0.2 U/ml thrombin to conduct plasma clot retraction analysis,which can be used to study the regulation of platelet function and evaluate the modulatory effects of drugs on plasma clot retraction.

关 键 词：血浆凝块收缩 凝血酶 CA^(2+) 血小板 信号通路抑制剂 

分 类 号：R446.1[医药卫生—诊断学]