Research Paper Volume 11, Issue 16 pp 6358—6370

Platelet activation in diabetic mice models: the role of vascular endothelial cell-derived protein disulfide isomerase-mediated GP IIb/IIIa receptor activation

Figure 6. Platelets of normal C57 mice were stimulated with EMP and pretreatment EMPs. Adenosine diphosphate (ADP) (10 μg/mL), RL90 (1 μg/mL), and rutin (60 μM) were used to pretreat EMPs obtained from four groups; then the EMP and the pretreatment EMP were used to stimulate the platelets of normal C57 mice, respectively. FCM was used to detect the level of platelet activation. EMP from the diabetic mice can significantly activate platelets, while RL90 and rutin could inhibit this process. Data were analyzed using one-way ANOVA. (A) Comparisons of the GP IIb/IIIa receptor expression level on the platelet surface of C57 mice stimulated using different pretreated EMPs, within each group. The GP IIb/IIIa receptor expression significantly increased in the EMP stimulus subgroup. *P < 0.05, **P < 0.01, ***P < 0.001 versus Control; #P < 0.05, ##P < 0.01, ###P < 0.001 versus EMP; P < 0.05, ††P < 0.01, P < 0.001 versus EMP + ADP (n = 3–5). (B) Comparisons of the expression level of P-selectin on the platelet surface of C57 mice stimulated using different pretreated EMPs, within each group. The P-selectin expression was significantly increased in the EMP stimulus subgroup. *P < 0.05, **P < 0.01, ***P < 0.001 versus Control; #P < 0.05, ##P < 0.01, ###P < 0.001 versus EMP; P < 0.05, ††P < 0.01, P < 0.001 versus EMP + ADP (n = 3–10). (C) Comparisons of the GP IIb/IIIa receptor expression level on the platelet surface of normal C57 mice among the four groups stimulated using different pretreated EMP. *P < 0.05, **P < 0.01,***P < 0.001 versus Chow; #P < 0.05, ##P < 0.01, ###P < 0.001 versus Chow + Rutin; P < 0.05, ††P < 0.01, P < 0.001 versus DM (n = 3–8). (D) Comparisons of the P-selectin expression level on the platelet surface of normal C57 mice among the four groups stimulated using different pretreated EMP. *P < 0.05, **P < 0.01, ***P < 0.001 versus Chow; #P < 0.05, ##P < 0.01, ###P < 0.001 versus Chow + Rutin; P < 0.05, ††P < 0.01, P < 0.001 versus DM (n = 3–9). (E) The Duolink® in-situ proximity ligation assay was used to detect the GPIIb/IIIa receptor and EMP-PDI on the platelet surface. The red dot granule represents GPIIb/IIIa receptor binding to PDI (scale = 50 μm).