Research Article

Collecting duct PGE2 responses reduce water loss with empagliflozin in mice with type 2 diabetes mellitus

R Nasrallah, J Zimpelmann, V Cheff, JF Thibodeau, KD Burns and RL Hébert*

Published: 04/08/2021 | Volume 5 - Issue 1 | Pages: 023-030


Introduction: Sodium-glucose cotransporter 2 inhibitors such as empagliflozin (EMPA) protect against diabetic kidney disease. Prostaglandin E2 (PGE2) the main renal product of cyclooxygenase-2, inhibits vasopressin (AVP)-water reabsorption in the collecting duct (CD). The novelty of this study is that for the first time, we examined if EMPA affects the renal PGE2/EP receptor system and determined if CD responses to EMPA prevent water loss.

Methods: Four groups of adult male mice were studied after 6 weeks of treatment: control (db/m), db/m+EMPA (10 mg/kg/day in chow), type 2 diabetic diabetic/dyslipidemia (db/db), and db/db+EMPA. Tubules were microdissected for quantitative polymerase chain reaction (qPCR) and CD water transport was measured in response to AVP, with or without PGE2.

Results: Hyperglycemia and albuminuria were attenuated by EMPA. Renal mRNA expression for COX, PGE synthase, PGE2 (EP) receptor subtypes, CD AVP V2 receptors and aquaporin-2 was elevated in db/db mice, but unchanged by EMPA. Urine PGE2 levels increased in db/db but were unchanged by EMPA. AVP-water reabsorption was comparable in db/m and db/m+EMPA, and equally attenuated to 50% by PGE2. In db/db mice, AVP-water reabsorption was reduced by 50% compared to non-diabetic mice, and this reduction was unaffected by EMPA. In db/db mice, AVP-stimulated water transport was more significantly attenuated with PGE2 (62%), compared to non-diabetic mice, but this attenuation was reduced in response to EMPA, to 28%.

Conclusion: In summary, expression of renal PGE2/EP receptors is increased in db/db mice, and this expression is unaffected by EMPA. However, in diabetic CD, PGE2 caused a greater attenuation in AVP-stimulated water reabsorption, and this attenuation is reduced by EMPA. This suggests that EMPA attenuates diabetes-induced excess CD water loss.

Read Full Article HTML DOI: 10.29328/journal.jcn.1001069 Cite this Article


  1. Rastogi A, Bhansali A. SGLT2 Inhibitors through the Windows of EMPA-REG and CANVAS Trials: A Review. Diabetes Ther. 2017; 8: 1245–1251. PubMed:
  2. Wanner C, Inzucchi SE, Lachin JM, Fitchett D, von Eynatten M, et al. Empagliflozin and Progression of Kidney Disease in Type 2 Diabetes. N Engl J Med. 2016; 375: 323-334.
  3. Heerspink HJL, Perkins BA, Fitchett DH, Husain M, Cherney DZI. Sodium Glucose Cotransporter 2 Inhibitors in the Treatment of Diabetes: Cardiovascular and Kidney Effects, Potential Mechanisms and Clinical Applications. Circulation 2016; 134: 752-772. PubMed:
  4. Cherney DZI, Perkins BA, Soleymanlou N, Maione M, Lai V, et al. Renal Hemodynamic Effect of Sodium-Glucose Cotransporter 2 Inhibition in Patients with Type 1 Diabetes Mellitus. Circulation. 2014; 129: 587-597.                         PubMed:
  5. Nasrallah R, Clark J, Hébert RL. Prostaglandins in the kidney: developments since Y2K. Clin Sci (Lond). Review. 2007; 113: 297-311. PubMed:
  6. Nasrallah R, Hassouneh R, Hébert RL. Chronic kidney disease: targeting prostaglandin E2 receptors. Am J Physiol. 307: 2014; F243-250. PubMed:
  7. Nasrallah R, Hassouneh R, Hébert RL. Prostaglandin E2, kidney disease, and cardiovascular risk: beyond hypertension and diabetes. J Am Soc Nephrol. 2016; 27: 666–676. PubMed:
  8. Nasrallah R, Xiong H, Hébert RL. Renal prostaglandin E2 receptor (EP) expression profile is altered in streptozotocin and B6-Ins2Akita type 1 diabetic mice. Am J Physiol. 2007; 292: 278–284. PubMed:
  9. Nasrallah R, Robertson SJ, Hébert RL. Chronic COX inhibition reduces diabetes-induced hyperfiltration, proteinuria, and renal pathological markers in 36-week B6-Ins2(Akita) mice. Am J Nephrol. 2009; 30: 346-353. PubMed:
  10. Nasrallah R, Robertson SJ, Karsh J, Hébert RL. Celecoxib modifies glomerular basement membrane, mesangial area and podocyte structure in OVE26 mice, but ibuprofen is more detrimental. Clin Sci. 2013; 124: 685-694. PubMed:
  11. Cherney DZI, Miller J, Scholey JW, Bradley TJ, Slorach C, et al. The effect of cyclooxygenase 2 inhibition on renal hemodynamic function in humans with type 1 diabetes mellitus. Diabetes 2008; 57: 688-695. PubMed:
  12. Thibodeau JF, Nasrallah R, Hébert RL, Kennedy C. Reduced albuminuria in diabetic PTGER1-null mice. Am J Pathol. 2013; 183: 1789-1802.
  13. Hassouneh R, Nasrallah R, Zimpelmann J, Gustol A, Ghossein J, et al. ProstaglandinE2/EP3 inhibits water reabsorption and contributes to polyuria and kidney injury in streptozotocin diabetic mice. Diabetologia. 2016; 59: 1318-1328. PubMed:
  14. Fu Y, Breljak D, Onishi A, Batz F, Patel R, et al. Organic anion transporter OAT3 enhances the glucosuric effect of the SGLT2 inhibitor empagliflozin. Am J Physiol. 2018; 315: F386-F394. PubMed:
  15. Nasrallah R, Zimpelmann J, Eckert D, Ghossein J, Geddes S, et al. Prostaglandin E2 EP1 receptor inhibits vasopressin-dependent water reabsorption and sodium transport in mouse collecting duct. Lab Invest. 2018; 98: 360-370.
  16. Nadler SP, Zimpelmann JA, Hébert RL. Prostaglandin E2 inhibits water permeability at a post-cAMP site in rat terminal inner medullary collecting duct. Am J Physiol. 1992; 262: F229-235. PubMed:
  17. Hébert RL, Jacobson HR, Breyer MD. Prostaglandin E2 inhibits sodium transport in the rabbit CCD by raising intracellular calcium. J. Clin. Invest. 1991; 87: 1992-1998. PubMed:
  18. Guan YY, Zhang RM. Breyer B, Fowler L, Davis RL, et al. Prostaglandin E2 inhibits renal collecting duct Na+ absorption by activating the EP1 receptor. J Clin Invest. 1998; 102: 194–201. PubMed:
  19. Zeni L, Norden AGW, Cancarini G, Unwin RJ. A more tubulocentric view of diabetic kidney disease. J Nephrol. 2017; 30: 701-717. PubMed:
  20. Vallon V, Platt KA, Cunard R, Schroth J, Whaley J, et al. SGLT2 mediates glucose reabsorption in the early proximal tubule. J Am Soc Nephrol. 2011; 22: 104-112. PubMed:
  21. Wang XX, Levi J, Luo Y, Myakala K, Herman-Edelstein M, et al. SGLT2 Expression is increased in Human Diabetic Nephropathy: SGLT2 Inhibition Decreases Renal Lipid Accumulation, Inflammation and the Development of Nephropathy in Diabetic Mice. J Biol Chem. 2017; 292: 5335–5348. PubMed:
  22. Jurczak MJ, Lee HY, Birkenfeld AL, Jornayvaz FR, Frederick DW, et al. SGLT2 Deletion Improves Glucose Homeostasis and Preserves Pancreatic b-Cell Function. Diabetes 2011; 60: 890–898. PubMed:
  23. Barnett AH, Mithal A, Manassie J, et al. Efficacy and safety of empagliflozin added to existing antidiabetes treatment in patients with type 2 diabetes and chronic kidney disease: a randomised, double-blind, placebo-controlled trial. Lancet Diabetes Endocrinol. 2014; 2: 369-384. PubMed:
  24. Gao M, Cao R, Du S, et al. Disruption of prostaglandin E2 receptor EP4 impairs urinary concentration via decreasing aquaporin 2 in renal collecting ducts. Proc Natl Aca Sci. 2015; 112: 8397-8402.
  25. Wang XX, Levi J, Luo Y, Myakala K, Herman-Edelstein M, et al. SGLT2 Expression is increased in Human Diabetic Nephropathy: SGLT2 Inhibition Decreases Renal Lipid Accumulation, Inflammation and the Development of Nephropathy in Diabetic Mice. J Biol Chem. 2017; 292: 5335–5348. PubMed:
  26. Terami N, Ogawa D, Tachibana H, Hatanaka T, Wada J, et al. Long-Term Treatment with the Sodium Glucose Cotransporter 2 Inhibitor, Dapagliflozin, Ameliorates Glucose Homeostasis and Diabetic Nephropathy in db/db Mice. PLoS ONE. 2014; 9: e100777. PubMed:
  27. Gallo LA, Ward MS, Fotheringham AK, Zhuang A, Borg DJ, et al. Once daily administration of the SGLT2 inhibitor, empagliflozin, attenuates markers of renal fibrosis without improving albuminuria in diabetic db/db mice. Sci Rep. 2016; 6: 26428. PubMed:
  28. Gangadharan KM, Gross S, Mudaliar H, Huang C, Pegg K, et al. Inhibition of kidney proximal tubular glucose reabsorption does not prevent against diabetic nephropathy in type 1 diabetic eNOS knockout mice. PLoS One. 2014; 9: e108994. PubMed:
  29. Vallon V, Gerasimova M, Rose MA, Masuda T, Satriano J, et al. SGLT2 inhibitor empagliflozin reduces renal growth and albuminuria in proportion to hyperglycemia and prevents glomerular hyperfiltration in diabetic Akita mice. Am J Physiol Renal Physiol. 2014; 306: F194–F204. PubMed:
  30. Nasrallah R, Hassouneh R, Zimpelmann J, Karam AJ, Thibodeau JF, et al. Prostaglandin E2 increases proximal tubule fluid reabsorption and modulates cultured proximal tubule cell responses via EP1 and EP4 receptors. Lab Invest. 2015; 95: 1044-1055. PubMed: