Sex-dependent effects of canagliflozin and dapagliflozin on hemostasis in normoglycemic and hyperglycemic mice

Abstract

Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are antihyperglycemic drugs that decrease mortality from cardiovascular diseases. However, their effects on hemostasis in the cardioprotective effects have not been evaluated. Therefore, the effects of canagliflozin (CANA, 100 mg/kg, p.o.) and dapagliflozin (DAPA, 10 mg/kg, p.o.) on the parameters of hemostasis were investigated in female and male normoglycemic and streptozotocin (180 mg/kg, i.p.)-induced diabetic mice. CANA and DAPA reduced platelet activity in thrombus in male and female mice both normoglycemic and diabetic. CANA decreased thrombus formation in diabetic male mice, and platelet activation to ADP in diabetic female and male mice. Activation of fibrinolysis was observed in female mice, both normoglycemic and diabetic. DAPA reduced thrombus formation in diabetic male and female mice, and decreased platelet activation to ADP and fibrin formation in diabetic male mice. DAPA increased fibrin formation in normoglycemic female mice and activated fibrinolysis in diabetic female mice. CANA and DAPA exerted sex-specific effects, which were more pronounced in hyperglycemia. The antithrombotic effect of CANA and DAPA was more noticeable in male mice and could be due to platelet inhibition. The effect on coagulation and fibrinolysis was not clear since an increased coagulation and fibrinolysis were observed only in female mice.

Figure 1

The effects of CANA and DAPA on laser-induced thrombosis. (A) The effects of DAPA and CANA on the thrombus area. (B) Representative confocal microscopy images of thrombi. Thrombus and the vessel wall have been indicated. Bar = 10 µm. Pictures were captured with SlideBook 6.0. ^#p < 0.05, ^###p < 0.001 vs Contr Norm, *p < 0.05, **p < 0.01 vs Contr STZ; n = 7–14.

Figure 2

The effects of CANA and DAPA on the PECAM-1/thrombus ratio. ^##p < 0.01, ^###p < 0.001 vs Contr Norm, ***p < 0.001 vs Contr STZ, ^p < 0.05 vs Female CANA STZ, ^$$p < 0.01 vs Male CANA Norm; n = 7–14.

Figure 3

Assessment of the platelet activation index using flow cytometry. (A) The effects of CANA and DAPA on the platelet activation index. (B) Representative images from flow cytometry, which show platelet populations after ADP treatment. Red arrows indicate the appearance of the P3 population (platelets with PS). Dot plots were captured with CytExpert 2.4. ^###p < 0.001 vs Contr Norm, *p < 0.05, ***p < 0.001 vs Contr STZ; n = 7–10.

Figure 4

The effects of CANA and DAPA on fibrin net density. (A) The effects of CANA and DAPA on fibrin net density in clots formed after recalcination of PPP. (B) Representative confocal microscopy images of fibrin net. Bar = 10 µm. Pictures were captured with SlideBook 6.0. ^##p < 0.01, ^###p < 0.001 vs Contr Norm, ***p < 0.001 vs Contr STZ, ^^^p < 0.001 vs Female Contr STZ; n = 7–9.

Figure 5

The effects of CANA and DAPA on euglobulin clot lysis time. ^#p < 0.05, ^###p < 0.001 vs Contr Norm, ***p < 0.001 vs Contr STZ, ^^p < 0.01 vs Female Contr Norm, ^$$p < 0.01 vs Female Contr STZ; n = 7–11.