Effects of single and dual RAAS blockade therapy on progressive kidney disease transition to CKD in rats

  • Devesh Aggarwal
  • Gaaminepreet SinghEmail author
Original Article


Ischemic reperfusion (I/R) is the primary cause of acute kidney injury (AKI) in hospitalized patients. Although AKI resolution occurs in few days, it predisposes kidneys to progressive renal injury. Previously, administration of rennin-angiotensin-aldosterone system (RAAS) blocker spironolactone in acute phase was reported to attenuate various manifestations of chronic kidney disease (CKD) in rats. The present study investigates the effects of RAAS blockade during progressive kidney disease (30 days onwards) on CKD outcomes in rodent model of I/R injury. CKD was induced by clamping both renal pedicles for 45 min followed by 90 days of reperfusion in rats. Single and dual RAAS blocker therapy was initiated at 30 days post-I/R injury and continued until the end of the study period. Evaluation of proteinuria and creatinine levels was done every 30 days in various study groups. Assessment of CKD was done by analyzing renal tissue oxidative stress, inflammatory biomarker levels, and histological changes after 90 days of I/R injury. After 90 days, I/R rat kidneys displayed hypertrophy, reduced body weight, increased oxidative stress, elevated inflammatory biomarker levels, and histological abnormalities such as glomerulosclerosis, mesangial expansion, and tubulointerstitial fibrosis. Treatment with losartan or spironolactone alone significantly reduced various CKD-associated features. Remarkably, combined treatment with dual RAAS blocker in low dose or high dose exhibited highest beneficial effects on various parameters in CKD model, with low-dose combination showing fewer side effects. Therefore, we propose that combined low-dose RAAS blockade therapy might serve as a better therapeutic approach for retarding progressive kidney disease transition to CKD.


Acute kidney injury Chronic kidney injury Renin angiotensin and aldosterone system Ischemia reperfusion 


Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the ISF College of Pharmacy, Moga, Punjab, India (ISFCP/IAEC/CPCSEA/Meeting No. 23/2018/Protocol No. 371).


  1. Aebi H (1974) Catalase. In: Methods of enzymatic analysis. Elsevier, pp 673–684Google Scholar
  2. Barrera-Chimal J, Pérez-Villalva R, Rodríguez-Romo R, Reyna J, Uribe N, Gamba G, Bobadilla NA (2013) Spironolactone prevents chronic kidney disease caused by ischemic acute kidney injury. Kidney Int 83:93–103. CrossRefPubMedGoogle Scholar
  3. Barrera-Chimal J, Rocha L, Amador-Martínez I et al (2019) Delayed spironolactone administration prevents the transition from acute kidney injury to chronic kidney disease through improving renal inflammation. Nephrol Dial Transplant 34:794–801. CrossRefPubMedGoogle Scholar
  4. Basile DP, Donohoe D, Roethe K, Osborn JL (2001) Renal ischemic injury results in permanent damage to peritubular capillaries and influences long-term function. Am J Physiol-Ren Physiol 281:F887–F899. CrossRefGoogle Scholar
  5. Basile DP, Leonard EC, Beal AG et al (2012) Persistent oxidative stress following renal ischemia-reperfusion injury increases ANG II hemodynamic and fibrotic activity. Am J Physiol-Ren Physiol 302:F1494–F1502. CrossRefGoogle Scholar
  6. Bedford M, Farmer C, Levin A, Ali T, Stevens P (2012) Acute kidney injury and CKD: chicken or egg? Am J Kidney Dis 59:485–491. CrossRefPubMedGoogle Scholar
  7. Bomback AS, Kshirsagar AV, Amamoo MA et al (2008) Change in proteinuria after adding aldosterone blockers to ACE inhibitors or angiotensin receptor blockers in CKD: a systematic review. Am J Kidney Dis 51:199–211CrossRefGoogle Scholar
  8. Boyne AF, Ellman GL (1972) A methodology for analysis of tissue sulfhydryl components. Anal Biochem 46:639–653. CrossRefPubMedGoogle Scholar
  9. Chawla LS, Kimmel PL (2012) Acute kidney injury and chronic kidney disease: an integrated clinical syndrome. Kidney Int 82:516–524CrossRefGoogle Scholar
  10. Cicoira M, Zanolla L, Rossi A, Golia G, Franceschini L, Cabrini G, Bonizzato A, Graziani M, Anker SD, Coats AJ, Zardini P (2001) Failure of aldosterone suppression despite angiotensin-converting enzyme (ACE) inhibitor administration in chronic heart failure is associated with ACE DD genotype. J Am Coll Cardiol 37:1808–1812CrossRefGoogle Scholar
  11. Coca SG, Singanamala S, Parikh CR (2012) Chronic kidney disease after acute kidney injury: a systematic review and meta-analysis. Kidney Int 81:442–448CrossRefGoogle Scholar
  12. Conger JD, Robinette JB, Hammond WS (1991) Differences in vascular reactivity in models of ischemic acute renal failure. Kidney Int 39:1087–1097. CrossRefPubMedGoogle Scholar
  13. Donnahoo KK, Meng X, Ayala A et al (1999) Early kidney TNF-α expression mediates neutrophil infiltration and injury after renal ischemia-reperfusion. Am J Physiol-RegulIntegr Comp Physiol 277:R922–R929. CrossRefGoogle Scholar
  14. Glynne PA, Picot J, Evans TJ (2001) Co expressed nitric oxide synthase and apical beta(1) integrins influence tubule cell adhesion after cytokine-induced injury. J Am Soc Nephrol 12(11):2370–2383PubMedGoogle Scholar
  15. Hirst JA, Taylor KS, Stevens RJ et al (2012) The impact of renin–angiotensin–aldosterone system inhibitors on Type 1 and Type 2 diabetic patients with and without early diabetic nephropathy. Kidney Int 81:674–683CrossRefGoogle Scholar
  16. Hirose R, Xu F, Dang K, Liu T, Behrends M, Brakeman PR, Wiener-Kronish J, Niemann CU (2008) Transient hyperglycemia affects the extent of ischemia-reperfusion-induced renal injury in rats. Anaesthesiology 108(3):402–414CrossRefGoogle Scholar
  17. Ivanov M, Mihailović-Stanojević N, Grujić Milanović J et al (2014) Losartan improved antioxidant defense, renal Function and structure of Postischemic Hypertensive Kidney. PLoS ONE 9:e96353. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Jefferson JA, Shankland SJ, Pichler RH (2008) Proteinuria in diabetic kidney disease: a mechanistic viewpoint. Kidney Int 74:22–36. CrossRefPubMedGoogle Scholar
  19. Junge W, Wilke B, Halabi A, Klein G (2004) Determination of reference intervals for serum creatinine, creatinine excretion and creatinine clearance with an enzymatic and a modified Jaffé method. Clin Chim Acta 344:137–148. CrossRefPubMedGoogle Scholar
  20. Kang YS, Ko GJ, Lee MH et al (2008) Effect of eplerenone, enalapril and their combination treatment on diabetic nephropathy in type II diabetic rats. Nephrol Dial Transplant 24:73–84. CrossRefPubMedGoogle Scholar
  21. Lee MY, Shim MS, Kim BH et al (2011) Effects of spironolactone and losartan on diabetic nephropathy in a type 2 diabetic rat model. Diabetes Metab J 35:130. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Lozano-Maneiro L, Puente-García A (2015) Renin-angiotensin-aldosterone system blockade in diabetic nephropathy. Present Evidences J Clin Med 4:1908–1937. CrossRefPubMedGoogle Scholar
  23. Malek M (2015) Renal ischemia/reperfusion injury; from pathophysiology to treatment. J Renal. Inj Prev.
  24. Medeiros M, Velásquez-Jones L, Hernández AM, Ramón-García G, Valverde S, Fuentes Y, Vargas A, Patiño M, Pérez-Villalva R, Ortega-Trejo JA, Barrera-Chimal J, Bobadilla NA (2017) Randomized controlled trial of mineralocorticoid receptor blockade in children with chronic kidney allograft nephropathy. Clin J Am Soc Nephrol 12:1291–1300. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358. CrossRefPubMedGoogle Scholar
  26. Rachmani R, Slavachevsky I, Amit M et al (2004) The effect of spironolactone, cilazapril and their combination on albuminuria in patients with hypertension and diabetic nephropathy is independent of blood pressure reduction: a randomized controlled study. Diabet Med 21(5):471–475CrossRefGoogle Scholar
  27. Ritz E (2003) Angiotensin II and oxidative stress: an unholy alliance. J Am Soc Nephrol 14:2985–2987. CrossRefPubMedGoogle Scholar
  28. Rodríguez-Romo R, Benítez K, Barrera-Chimal J, Pérez-Villalva R, Gómez A, Aguilar-León D, Rangel-Santiago JF, Huerta S, Gamba G, Uribe N, Bobadilla NA (2016) AT1 receptor antagonism before ischemia prevents the transition of acute kidney injury to chronic kidney disease. Kidney Int 89:363–373. CrossRefPubMedGoogle Scholar
  29. Rüster C, Wolf G (2006) Renin-angiotensin-aldosterone system and progression of renal disease. J Am Soc Nephrol 17:2985–2991. CrossRefPubMedGoogle Scholar
  30. Sato A, Saruta T (2001) Aldosterone escape during angiotensin converting enzyme inhibitor therapy in essential hypertensive patients with left ventricular hypertrophy. J Int Med Res 29:13–21. CrossRefPubMedGoogle Scholar
  31. Savory J, Pu PH, Sunderman FW (1968) A biuret method for determination of protein in normal urine. Clin Chem 14(12):1160–1171PubMedGoogle Scholar
  32. Schjoedt KJ, Andersen S, Rossing P, Tarnow L, Parving HH (2004) Aldosterone escape during blockade of the renin–angiotensin–aldosterone system in diabetic nephropathy is associated with enhanced decline in glomerular filtration rate. Diabetologia 47(11):1936–1939CrossRefGoogle Scholar
  33. Singh G, Krishan P (2018) Cobalt treatment does not prevent glomerular morphological alterations in type 1 diabetic rats. Naunyn Schmiedebergs Arch Pharmacol 391:933–944. CrossRefPubMedGoogle Scholar
  34. Staessen J, Lijnen P, Fagard R (1981) Rise in plasma concentration of aldosterone during long-term angiotensin II suppression. J Endocrinol 91(3):457–465CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of PharmacologyISF College of PharmacyMogaIndia

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