A Novel Flow Cytometry based Mouse Platelet Aggregation Method and Its Application for Accessing the Effects of Tetrameth-ylpyrazine
Mei Jinping
Zhou Xin
Ji Wenjie
Ma Yongqiang
Yang Guohong
Guo Zhaozeng
Li Yuming
Zhao Jihong
Abstract：Objective To evaluate a novel method based on flow cytometry to assess mouse platelet aggregation and the effects of tetramethylpyrazine (TMPZ)on mouse platelet.Methods Platelets from the same mouse whole blood context (n=5)were labeled with CD61 PE or CD61 FITC,and were analyzed for the formation of double colored aggregates by flow cytometry after mixing and upon appropriate stimulation with ADP (20μmol/L).16 male C57BL/6J mice were randomly divided into 2 groups of normal saline group and clopidogrel group.The platelet aggregation was measured by FCM after 7 days of drug administration.Whole C57BL/6J mouse blood intervened by TMPZ was divided into 3 subgroups:low (0.5 mg/mL),middle (1 mg/mL)and high concentration (2 mg/mL)and tirofiban (5μg/mL)group.Mouse platelet aggregation was tested by FCM.Results Upon stimulation with ADP the percent-age of double colored events increased over time and their forward scatter/side scatter positioning shifted significantly compared with single platelets (P<0.01).Mouse platelet aggregation after oral clopidogrel was significantly reduced from (19.57± 1.12)% to (8.71± 1.21)% (P<0.05).Mouse whole blood was intervened by low,middle and high doses of TMPZ and tirofiban,and the mouse platelet aggregation rate dropped significantly compared with that in the normal saline group (P<0.05).Conclusion The FCM based method of evaluating platelet aggregation can show the activities of P2Y12 receptor antagonist,GPⅡb/Ⅲa receptor antagonist and TMPZ,and this is potential y very relevant for both the study of platelet and for the screening studies of the develop-ment of novel hemostatic drugs.
Keywords：platelet aggregationflow cytometrymousetetramethylpyrazine
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 585-588 )