Blocking mineralocorticoid signaling with esaxerenone reduces atherosclerosis in hyperglycemic ApoE KO mice without affecting blood pressure and glycolipid metabolism

Abstract

Endothelial damage mediated by mineralocorticoid receptor (MR) is an important factor in the development of atherosclerosis. Esaxerenone is a highly selective drug that can specifically block MR activity. The aim of this study is to examine whether specific blocking of mineralocorticoid signaling with esaxerenone exerts favorable effects on the progression of atherosclerosis. ApoE KO mice were used as a model of atherosclerosis. In addition to a non-diabetic model, we created a diabetic model using streptozotocin. These were divided into a control group and an esaxerenone group. Esaxerenone-containing diet was provided for 8 weeks starting at 10 weeks of age. Various metabolic markers and abdominal aortic mRNA expression were evaluated, and histological examination of the aortic arch and thoracic aorta was performed. We also used human aortic smooth muscle cells (HASMCs) to investigate the possible direct effects of esaxerenone on vascular smooth muscle cells. In diabetic mice, plaque area in the aortic arch was significantly smaller in esaxerenone group compared to control group, although there were no differences in blood pressure, serum lipid levels between the two groups. Inflammation-related genes, macrophage marker, cell adhesion factors and oxidative stress marker were all significantly lower in esaxerenone group. The studies using HASMCs have confirmed that esaxerenone has anti-inflammatory effects on vascular smooth muscle cells. Specific blocking of mineralocorticoid signaling with esaxerenone exerts favorable effects on the progression of atherosclerosis without influencing blood pressure and glycolipid metabolism.

Keywords: Atherosclerosis; Esaxerenone; Oxidative stress.

Fig. 1

There was no significant difference between esaxerenone and control group in (A, B) non-fasting blood glucose levels (A, B), body weights (C, D), and food intake in non-diabetic and diabetic mice (E, F) (n = 9–16). Black, control group; white, esaxerenone group.

Fig. 2

Atherosclerotic lesion in aortic arch in non-DM Control (A), non-DM Esaxerenone (B), DM Control (C) and DM Esaxerenone (D). (E) Esaxerenone treatment significantly reduced plaque area compared to control group in diabetic mice. n = 8. *: p < 0.05.

Fig. 3

(A) In esaxerenone group, urinary 8-OHdG levels were significantly lower compared to control in diabetic mice. n = 8. (B–D) mRNA expression levels of inflammatory markers in the abdominal aorta were lower in esaxerenone group compared to control in diabetic mice. n = 12. *: p < 0.05.

Fig. 4

(A–C) mRNA expression levels related to coagulation such as PAI-1, cell adhesion molecules such as V-CAM and I-CAM were significantly lower in esaxerenone group compared to control in diabetic mice. (D) Expression level of a macrophage marker F4/80 was not different between control and esaxerenone group. (E) Expression level of CD68 was significantly lower in esaxerenone group compared to control in diabetic mice. (F) Expression level of a plaque stability marker TIMP-1 was not different between the control and esaxerenone group. (G) Expression levels MMP-2 was significantly lower in esaxerenone group compared to control in diabetic mice. n = 12. *: p < 0.05.

Fig. 5

(A, B) The mRNA expression levels of inflammatory markers in human aortic smooth muscle cells (HASMCs) were significantly lower in the group treated with 100 nM of esaxerenone than in the group treated with LPS alone. (C, D) The mRNA expression levels of V-CAM and I-CAM, which are related to cell adhesion molecules, were also significantly lower in the group treated with 100 nM of esaxerenone than in the group treated with LPS alone. Con: control. LPS: stimulated with 100 ng/mL of LPS. LPS + Esa10: 10 nM of esaxerenone was administered together with LPS stimulation. LPS + E100: 100 nM of esaxerenone was administered together with LPS stimulation. n = 3. #: p < 0.05, Con versus LPS. *: p < 0.05, LPS versus LPS + Esa100.