Diabetes Metab J.  2020 Feb;44(1):186-192. 10.4093/dmj.2018.0271.

Evogliptin, a Dipeptidyl Peptidase-4 Inhibitor, Attenuates Renal Fibrosis Caused by Unilateral Ureteral Obstruction in Mice

• Kim MJ
• Kim NY
• Jung YA
• Lee S
• Jung GS
• Kim JG
• Lee IK
• Lee S
• Choi YK
• Park KG

• KMID: 2470966
• DOI: http://doi.org/10.4093/dmj.2018.0271

Abstract

Renal fibrosis is considered to be the final common outcome of chronic kidney disease. Dipeptidyl peptidase-4 (DPP-4) inhibitors have demonstrated protective effects against diabetic kidney disease. However, the anti-fibrotic effect of evogliptin, a DPP-4 inhibitor, has not been studied. Here, we report the beneficial effects of evogliptin on unilateral ureteral obstruction (UUO)-induced renal fibrosis in mice. Evogliptin attenuated UUO-induced renal atrophy and tubulointerstitial fibrosis. Immunohistochemistry and Western blotting demonstrated that evogliptin treatment inhibits pro-fibrotic gene expressions and extracellular matrix production. In vitro findings showed that the beneficial effects of evogliptin on renal fibrosis are mediated by inhibition of the transforming growth factor-β/Smad3 signaling pathway. The present study demonstrates that evogliptin is protective against UUO-induced renal fibrosis, suggesting that its clinical applications could extend to the treatment of kidney disease of non-diabetic origin.

Keyword

Dipeptidyl-peptidase IV inhibitors; Kidney failure, chronic; Transforming growth factor beta

MeSH Terms

Animals
Atrophy
Blotting, Western
Diabetic Nephropathies
Dipeptidyl-Peptidase IV Inhibitors
Extracellular Matrix
Fibrosis*
Gene Expression
Immunohistochemistry
In Vitro Techniques
Kidney Diseases
Kidney Failure, Chronic
Mice*
Renal Insufficiency, Chronic
Transforming Growth Factor beta
Ureter*
Ureteral Obstruction*
Dipeptidyl-Peptidase IV Inhibitors
Transforming Growth Factor beta

Figure

• 

  Fig. 1 Effects of evogliptin (Evo) on renal fibrosis and pro-fibrotic gene expression in kidneys of unilateral ureteral obstruction (UUO) mice. A UUO kidney from a mouse at day 11. (A) Representative images of hematoxylin and eosin (H&E) and Sirius red staining of kidney tissue sections from control (CON) mice and UUO mice treated with (300 mg/kg) or without Evo. The number of atrophic tubules was determined by measuring the amount of abnormal irregular and dilated tubular basement membranes in H&E-stained sections under high power magnification (×200). Areas of positive staining with Sirius red were quantitated by computer-based morphometric analysis. All morphometric data were normalized against the corresponding val-ues in CON animals. Data in all bar graphs are expressed as fold increases relative to the CON (n=6 in each group). (B) Representative images of immunohistochemical staining for phosphorylated-Smad3 (p-Smad3), plasminogen activator inhibitor 1 (PAI-1), fibronectin, α-smooth muscle actin (α-SMA), and type I collagen in kidney tissue sections from CON mice or UUO mice treated with (300 mg/kg) or without Evo (n=6 in each group). Areas of positive staining with p-Smad3, PAI-1, fibronectin, α-SMA, and type I collagen antibodies were quantitated by computer-based morphometric analysis. Data are the mean±standard error of the mean (SEM) of five random fields from each kidney. (C) Representative western blot analysis of p-Smad3, total-Smad3 (t-Smad3), PAI-1, fibronectin, α-SMA, and type I collagen protein level in UUO kidneys from mice treated with (300 mg/kg) or without Evo (n=6 in each group). Data in the bar graphs are the mean±SEM. NS, not significant. aP<0.05, bP<0.01, cP<0.001.

• 

  Fig. 2 Effects of evogliptin (Evo) on transforming growth factor-β (TGF-β)-induced pro-fibrotic gene expression in cultured kidney cell lines. Representative Western blot analysis of phosphorylated-Smad3 (p-Smad3), total-Smad3 (t-Smad3), plasminogen activator inhibitor 1 (PAI-1), fibronectin, α-smooth muscle actin (α-SMA), and type I collagen protein level in TGF-β-stimulated (A) human proximal renal tubular epithelial (HK-2) cells and (B) normal rat kidney fibroblasts (NRK-49F) cells. Cells were incubated with TGF-β (5 ng/mL) treated with (100 or 200 µg/mL) or without Evo co-treatment for 24 hours. Data are the mean±standard error of the mean (SEM) of three independent measurements. (C) Effects of Evo on PAI-1 promoter activity in NRK-49F and NRK-52E cells. Cells were treated with TGF-β (5 ng/mL) treated with (100 µg/mL) or without Evo co-treatment for 24 hours. Data are the mean±SEM of three independent measurements. NS, not significant; Luc, luciferase. aP<0.05, bP<0.01, cP<0.001.

Reference

1. Atkins RC, Zimmet P. IFKF World Kidney Day 2010 Steering Committee. Diabetes: diabetic kidney disease: act now or pay later. Nat Rev Nephrol. 2010; 6:134–136.

2. Meng XM, Nikolic-Paterson DJ, Lan HY. TGF-β: the master regulator of fibrosis. Nat Rev Nephrol. 2016; 12:325–338.

3. Lan HY, Chung AC. TGF-β/Smad signaling in kidney disease. Semin Nephrol. 2012; 32:236–243.

4. Salvo F, Moore N, Arnaud M, Robinson P, Raschi E, De Ponti F, Begaud B, Pariente A. Addition of dipeptidyl peptidase-4 inhibitors to sulphonylureas and risk of hypoglycaemia: systematic review and meta-analysis. BMJ. 2016; 353:i2231.

5. Hirakawa H, Zempo H, Ogawa M, Watanabe R, Suzuki J, Akazawa H, Komuro I, Isobe M. A DPP-4 inhibitor suppresses fibrosis and inflammation on experimental autoimmune myocarditis in mice. PLoS One. 2015; 10:e0119360.

6. Kaji K, Yoshiji H, Ikenaka Y, Noguchi R, Aihara Y, Douhara A, Moriya K, Kawaratani H, Shirai Y, Yoshii J, Yanase K, Kitade M, Namisaki T, Fukui H. Dipeptidyl peptidase-4 inhibitor attenuates hepatic fibrosis via suppression of activated hepatic stellate cell in rats. J Gastroenterol. 2014; 49:481–491.

7. Suzuki T, Tada Y, Gladson S, Nishimura R, Shimomura I, Karasawa S, Tatsumi K, West J. Vildagliptin ameliorates pulmonary fibrosis in lipopolysaccharide-induced lung injury by inhibiting endothelial-to-mesenchymal transition. Respir Res. 2017; 18:177.

8. Eun Lee J, Kim JE, Lee MH, Song HK, Ghee JY, Kang YS, Min HS, Kim HW, Cha JJ, Han JY, Han SY, Cha DR. DA-1229, a dipeptidyl peptidase IV inhibitor, protects against renal injury by preventing podocyte damage in an animal model of progressive renal injury. Lab Invest. 2016; 96:547–560.

9. Tan X, Hu J. Evogliptin: a new dipeptidyl peptidase inhibitor for the treatment of type 2 diabetes. Expert Opin Pharmacother. 2016; 17:1285–1293.

10. Min HS, Kim JE, Lee MH, Song HK, Kang YS, Lee MJ, Lee JE, Kim HW, Cha JJ, Chung YY, Hyun YY, Han JY, Cha DR. Dipeptidyl peptidase IV inhibitor protects against renal interstitial fibrosis in a mouse model of ureteral obstruction. Lab Invest. 2014; 94:598–607.

11. Kim MK, Chae YN, Kim HD, Yang EK, Cho EJ, Choi SH, Cheong YH, Kim HS, Kim HJ, Jo YW, Son MH, Kim SH, Shin CY. DA-1229, a novel and potent DPP4 inhibitor, improves insulin resistance and delays the onset of diabetes. Life Sci. 2012; 90:21–29.

12. Jung GS, Kim MK, Jung YA, Kim HS, Park IS, Min BH, Lee KU, Kim JG, Park KG, Lee IK. Clusterin attenuates the development of renal fibrosis. J Am Soc Nephrol. 2012; 23:73–85.

13. Iwano M, Plieth D, Danoff TM, Xue C, Okada H, Neilson EG. Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest. 2002; 110:341–350.

14. Sato M, Muragaki Y, Saika S, Roberts AB, Ooshima A. Targeted disruption of TGF-beta1/Smad3 signaling protects against renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction. J Clin Invest. 2003; 112:1486–1494.

15. Wang D, Zhang G, Chen X, Wei T, Liu C, Chen C, Gong Y, Wei Q. Sitagliptin ameliorates diabetic nephropathy by blocking TGF-β1/Smad signaling pathway. Int J Mol Med. 2018; 41:2784–2792.

16. Uchida T, Oda T, Matsubara H, Watanabe A, Takechi H, Oshima N, Sakurai Y, Kumagai H. Renoprotective effects of a dipeptidyl peptidase 4 inhibitor in a mouse model of progressive renal fibrosis. Ren Fail. 2017; 39:340–349.

17. Jung GS, Jeon JH, Choe MS, Kim SW, Lee IK, Kim MK, Park KG. Renoprotective effect of gemigliptin, a dipeptidyl peptidase-4 inhibitor, in streptozotocin-induced type 1 diabetic mice. Diabetes Metab J. 2016; 40:211–221.

18. Shi S, Srivastava SP, Kanasaki M, He J, Kitada M, Nagai T, Nitta K, Takagi S, Kanasaki K, Koya D. Interactions of DPP-4 and integrin β1 influences endothelial-to-mesenchymal transition. Kidney Int. 2015; 88:479–489.

19. Liu WJ, Xie SH, Liu YN, Kim W, Jin HY, Park SK, Shao YM, Park TS. Dipeptidyl peptidase IV inhibitor attenuates kidney injury in streptozotocin-induced diabetic rats. J Pharmacol Exp Ther. 2012; 340:248–255.

20. Kanasaki K, Shi S, Kanasaki M, He J, Nagai T, Nakamura Y, Ishigaki Y, Kitada M, Srivastava SP, Koya D. Linagliptin-mediated DPP-4 inhibition ameliorates kidney fibrosis in streptozotocin-induced diabetic mice by inhibiting endothelial-to-mesenchymal transition in a therapeutic regimen. Diabetes. 2014; 63:2120–2131.