SGLT2 inhibition with empagliflozin attenuates myocardial oxidative stress and fibrosis in diabetic mice heart

Chenguang Li, Jie Zhang, Mei Xue, Xiaoyu Li, Fei Han, Xiangyang Liu, Linxin Xu, Yunhong Lu, Ying Cheng, Ting Li, Xiaochen Yu, Bei Sun, Liming Chen, Chenguang Li, Jie Zhang, Mei Xue, Xiaoyu Li, Fei Han, Xiangyang Liu, Linxin Xu, Yunhong Lu, Ying Cheng, Ting Li, Xiaochen Yu, Bei Sun, Liming Chen

Abstract

Background: Hyperglycaemia associated with myocardial oxidative stress and fibrosis is the main cause of diabetic cardiomyopathy. Empagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor has recently been reported to improve glycaemic control in patients with type 2 diabetes in an insulin-independent manner. The aim of this study was to investigate the effect of empagliflozin on myocardium injury and the potential mechanism in type 2 diabetic KK-Ay mice.

Methods: Thirty diabetic KK-Ay mice were administered empagliflozin (10 mg/kg/day) by oral gavage daily for 8 weeks. After 8 weeks, heart structure and function were evaluated by echocardiography. Oxidants and antioxidants were measured and cardiac fibrosis was analysed using immunohistochemistry, Masson's trichrome stain and Western blot.

Results: Results showed that empagliflozin improved diabetic myocardial structure and function, decreased myocardial oxidative stress and ameliorated myocardial fibrosis. Further study indicated that empagliflozin suppressed oxidative stress and fibrosis through inhibition of the transforming growth factor β/Smad pathway and activation of Nrf2/ARE signaling.

Conclusions: Glycaemic control with empagliflozin significantly ameliorated myocardial oxidative stress injury and cardiac fibrosis in diabetic mice. Taken together, these results indicate that the empagliflozin is a promising agent for the prevention and treatment of diabetic cardiomyopathy.

Keywords: Empagliflozin; Myocardial fibrosis; Oxidative stress; SGLT2; Type 2 diabetes mellitus.

Figures

Fig. 1
Fig. 1
Left ventricular echocardiographic representative images
Fig. 2
Fig. 2
Effect of empagliflozin on oxidative stress in the cardiac tissue homogenate. Lipid hydroperoxide (a), glutathione peroxidase (b), superoxide dismutase (c), malondialdehyde (d), Western blotting analysis of NOX4 in the mice myocardium (e, f). Data are expressed as the mean ± SD. *P < 0.05 vs. Con; #P < 0.05 vs. DM
Fig. 3
Fig. 3
Empagliflozin suppresses matrix accumulation and myocardial fibrosis in DM mice. Immunostaining of TGF-β1, collagen I and collagen III protein expression and Masson’s trichrome staining of the myocardium (a). The percentages of positive areas of TGF-β1 (b), collagen I (c), collagen III (d) and connective tissue fraction (e). Data are expressed as the mean ± SD. *P < 0.05 vs. Con; #P < 0.05 vs. DM
Fig. 4
Fig. 4
Effect of empagliflozin on Nrf2/ARE and TGF-β/SMAD pathway in vivo. a Western blot analysis for the expression of TGF-β/SMAD pathway was performed on protein isolated from the hearts of the three groups. b Western blot analysis for the expression of Nrf2/ARE pathway of heart tissue in mice. c The relative protein levels were calculated and Lamin B was used as an internal control. d The relative protein levels were calculated and β-actin was used as an internal control. Data are expressed as the mean ± SD. *P < 0.05 vs. Con; #P < 0.05 vs. DM

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