Characteristics and laboratory data from three groups of diabetic <i>db/db</i> mice with or without ipragliflozin, and wild-type mice that received vehicle.

<p>Non-diabetic wild-type mice that received vehicle, diabetic <i>db/db</i> mice that received vehicle, diabetic <i>db/db</i> mice that received ipragliflozin were measured. The values show mean ± SEM. Ipragliflozin, sodium glucose co-transporter 2 inhibitor.</p><p>One-way ANOVA followed by Tukey test:</p><p>^a,<i>p</i> < 0.05 vs. wild-type mice that received vehicle;</p><p>^b,<i>p</i> < 0.05 vs. diabetic <i>db/db</i> mice that received ipragliflozin.</p><p>Characteristics and laboratory data from three groups of diabetic <i>db/db</i> mice with or without ipragliflozin, and wild-type mice that received vehicle.</p

Correlation between foam cell formation and glycemic control in diabetic <i>Apoe</i>^{<i>−/−</i>} mice (A) or diabetic <i>db/db</i> mice (B).

<p>Fig 5A shows the correlation between foam cell formation and HbA1c in non-diabetic and diabetic <i>Apoe</i>^{<i>−/−</i>} mice. Diabetic mice that received vehicle (n = 10); r = 0.77, <i>p</i> < 0.01. Diabetic mice that received dapagliflozin (n = 5); r = 0.98, <i>p</i> < 0.005. Combined diabetic mice (n = 15); r = 0.91, <i>p</i> < 0.0001. Fig 5B shows the correlation between foam cell formation and HbA1c in diabetic <i>db/db</i> mice and wild-type (C57/BL6) mice. Diabetic mice that received vehicle (n = 18); r = 0.71, <i>p</i> < 0.001. Diabetic mice that received ipragliflozin (n = 13); r = 0.87, <i>p</i> < 0.0001. Combined diabetic mice (n = 31); r = 0.88, <i>p</i> < 0.0001. <i>r</i> values indicate Pearson correlation coefficients. Pearson’s correlation test, <i>p</i> < 0.05.</p

Changes in gene expression related to foam cell formation in the macrophages obtained from <i>Apoe</i>^{<i>−/−</i>} mice.

<p>Gene expressions were measured in the peritoneal macrophages obtained from non-diabetic <i>Apoe</i>^{<i>−/−</i>} mice (n = 7), diabetic <i>Apoe</i>^{<i>−/−</i>} mice that received vehicle (n = 6), and those that received dapagliflozin (n = 5). Gene expressions of (A) lectin-like ox-LDL receptor-1 (Lox-1), (B) CD36, (C) acyl-coenzyme A:cholesterol acyltransferase 1 (ACAT1), (D) ATP-binding cassette transporter A1 (ABCA1), and (E) ATP-binding cassette sub-family G member1 (ABCG1) and the association with GAPDH were analyzed by real-time RT-PCR before the addition of ox-LDL. The values show mean ± SEM. One-way ANOVA followed by Tukey test for the comparison of <i>Apoe</i>^{<i>−/−</i>} mice: *<i>p</i> < 0.05, ^†<i>p</i> < 0.01.</p

Foam cell formation in exudate peritoneal macrophages obtained from non-diabetic and diabetic <i>Apoe</i>^{<i>−/−</i>} mice (A), and diabetic <i>db/db</i> mice (B).

<p>Four days after an intraperitoneal injection of thioglycolate, the exudated peritoneal cells were isolated from the treated non-diabetic and diabetic <i>Apoe</i>^{<i>−/−</i>} mice at 21 weeks of age (Fig 3A), or from the diabetic <i>db/db</i> mice at 13 weeks of age (Fig 3B). Adherent macrophages were incubated for 18 hours with the RPMI-1640 medium containing 10μg/mL oxidized low-density lipoprotein (LDL) in the presence of 0.1 mmoL [³H]olate that was conjugated with bovine serum albumin. The cellular lipids were extracted and the radioactivity of the cholesterol [³H]olate was determined with thin-layer chromatography. Foam cell formation was expressed as cholesteryl ester (CE) accumulation. The values show mean ± SEM. One-way ANOVA followed by Tukey test: ^‡<i>p</i> < 0.001, ^§<i>p</i> < 0.0001.</p

Suppressive effects of dapagliflozine administration against the development of aortic atherosclerotic lesions in non-diabetic and diabetic <i>Apoe</i>^{<i>−/−</i>} mice.

<p>Twenty-two mice at 15 weeks of age were made diabetic with peritoneal injections of STZ (50 mg/kg/day) for 5 consecutive days and twenty mice were treated with saline. The 17-week-old non-diabetic and diabetic <i>Apoe</i>^{<i>−/−</i>} mice were orally given SGLT2i (dapagliflozin) or vehicle for 4 weeks, starting from 17 weeks of age. Representative atherosclerotic lesions in the aortic surface stained with oil red O (a-d) and measured (m). Yellow arrows show notable atherosclerotic lesions. In the aortic root, the atheromatous plaques and monocyte/macrophage accumulations were stained with Oil red O (e-h) or anti-MOMA2 antibody (i-l). Black arrows show notable atheromatous plaques. The severity of atheromatous plaques (n) and degree of monocyte/macrophage accumulation (o) were evaluated. The data are expressed as mean ± SEM. One way ANOVA followed by Tukey test: ^†<i>p</i> < 0.01, ^‡<i>p</i> < 0.001, ^§<i>p</i> < 0.0001.</p

Oral glucose tolerance test.

<p>Glucose (0.5 g/kg body weight) was administered orally through a gavage tube after 6 h of fasting, and blood glucose levels were measured at the specified time points of 0 (pre-glucose/fasting glucose level), and at 15, 30, 60, and 120 min after administration. Glucose curve after oral glucose loading in non-diabetic and diabetic apolipoprotein E-null mice (<i>Apoe</i>^{<i>−/−</i>}) mice that received vehicle or dapagliflozin (A), and the area under the curve (AUC) (B). Glucose curve after oral glucose loading in diabetic <i>db/db</i> mice that received vehicle or ipragliflozin (C), and the AUC (D). The data are expressed as mean ± SEM. One-way analysis of variance (ANOVA) followed by Tukey test for the comparison of <i>Apoe</i>^{<i>−/−</i>} mice: a, <i>p</i> < 0.05 vs. non-diabetic <i>Apoe</i>^{<i>−/−</i>} mice that received vehicle; b, <i>p</i> < 0.05 vs. non-diabetic <i>Apoe</i>^{<i>−/−</i>} mice that received dapagliflozin; c, <i>p</i> < 0.05 vs. diabetic <i>Apoe</i>^{<i>−/−</i>} mice that received dapagliflozin. Unpaired t-test for the comparison of <i>db/db</i> mice: d, <i>p</i> < 0.05 vs. vehicle. n = 5 per group. ^†<i>p</i> < 0.01, ^‡<i>p</i> < 0.001.</p

Changes in gene expression related to foam cell formation in the macrophages obtained from <i>db/db</i> mice and wild-type mice (C57/BL6).

<p>Gene expressions were measured in the peritoneal macrophages obtained from wild-type (C57/BL6) mice (n = 3), diabetic <i>db/db</i> mice that received vehicle (n = 12), and those that received ipragliflozin (n = 6). Gene expressions of (A) Lox-1, (B) CD36, (C) ACAT1, (D) ABCA1, and (E) ABCG1 in association with GAPDH were analyzed by real-time RT-PCR before the addition of ox-LDL. The values show mean ± SEM. One-way ANOVA followed by Tukey test: *<i>p</i> < 0.05, ^†<i>p</i> < 0.01.</p

MOESM1 of The role of endothelial nitric oxide in the anti-restenotic effects of liraglutide in a mouse model of restenosis

Additional file 1. Additional figures (Figs. S1–S4)

Surface atherosclerotic lesions of the aorta in 21-week-old nondiabetic <i>Apoe</i>^{−<b>/</b>−} mice treated without/with vildagliptin infused with incretin receptor blockers.

<p>During the 4-week period of vildagliptin administration, the <i>Apoe</i>^−/− mice (17-weeks-old at the outset) were respectively infused with saline (vehicle), GLP-1 receptor blocker (Ex-9), the GIPR blocker (Pro³), and Ex-9+Pro³ by osmotic mini-pump. The entire aorta was stained with Oil Red O.</p

Expressions of GIPR in various mouse tissues or cells.

<p>Relative gene expressions of GLP-1R and GIP in various mouse tissues or cells determined by real-time PCR using 10 and 11 primers when the gene expression of the vasculature (aorta) is settled as 1.000.</p