Molecular and Cellular Biochemistry

, Volume 451, Issue 1–2, pp 185–196 | Cite as

Modulatory effect of 4-phenyl butyric acid on hyperoxaluria-induced renal injury and inflammation

  • Minu Sharma
  • Amarjit S. Naura
  • S. K. SinglaEmail author


Hyperoxaluria-associated deposition of calcium oxalate crystals results from oxalate-induced renal injury and inflammation. The present study was designed to evaluate the effect of 4-Phenyl butyric acid (4-PBA), a chemical chaperone, in ethylene glycol-induced hyperoxaluria and compare its effect with antioxidant, N-acetyl cysteine (NAC). Male Sprague–Dawley rats were given ethylene glycol in drinking water for 28 days to induce hyperoxaluria. 4-PBA and NAC were given by oral gavage. Effect of 4-PBA was analyzed in both prophylactic and curative regimens. After every 7 days, 24-h urine samples were analyzed for kidney injury and inflammation markers. Increased amounts of kidney injury markers like Kidney injury molecule-1, Lactate dehydrogenase, and N-acetyl-β-glucoseaminidase were found in the urine of hyperoxaluric rats which were significantly reduced by 4-PBA treatment in both prophylactic and curative regimens. Inflammatory markers IL-1β, IL-6, and MCP-1 were also raised in the urine of hyperoxaluric rats which were significantly decreased by 4-PBA treatment. Hyperoxaluria was accompanied with renal oxidative stress as reflected by decreased glutathione redox status and increased reactive oxygen species which was significantly reduced by 4-PBA treatment. Histological study with H&E and Pizzolato staining showed numerous calcium oxalate crystal deposits in the renal tissues of hyperoxaluric rats. However, no significant crystal deposits were seen in the 4-PBA-treated hyperoxaluric rats. N-acetyl cysteine treatment effectively decreased renal oxidative stress but did not alter the production of inflammatory markers. Collectively, the present study suggested the potential protective effect of 4-PBA in hyperoxaluria-induced renal injury and inflammation.


Kidney Hyperoxaluria Nephrolithiasis 4-Phenyl butyric acid N-acetyl cysteine Oxidative stress 



The financial assistance provided by the Science and Engineering Research Board (SERB), Government of India, New Delhi is gratefully acknowledged.

Compliance with ethical standards

Conflict of interest

The authors state no conflict of interest.


  1. 1.
    Nazzal L, Puri S, Goldfarb DS (2016) Enteric hyperoxaluria: an important cause of end-stage kidney disease. Nephrol Dial Transplant 31:375–382. CrossRefPubMedGoogle Scholar
  2. 2.
    Roudakova K, Monga M (2014) The evolving epidemiology of stone disease. Indian J Urol 30:44–48. CrossRefPubMedGoogle Scholar
  3. 3.
    O’Kell AL, Grant DC, Khan SR (2017) Pathogenesis of calcium oxalate urinary stone disease: species comparison of humans, dogs, and cats. Urolithiasis 45:329–336. CrossRefPubMedGoogle Scholar
  4. 4.
    Asplin JR (2015) The management of patients with enteric hyperoxaluria. Urolithiasis. PubMedGoogle Scholar
  5. 5.
    Convento MB, Pessoa EA, Cruz E, da Gloria MA, Schor N, Borges FT (2017) Calcium oxalate crystals and oxalate induce an epithelial-to-mesenchymal transition in the proximal tubular epithelial cells: contribution to oxalate kidney injury. Sci Rep 7:45740. CrossRefPubMedGoogle Scholar
  6. 6.
    Khan SR (2014) Reactive oxygen species, inflammation and calcium oxalate nephrolithiasis. Transl Androl Urol 3:256–276. PubMedGoogle Scholar
  7. 7.
    Davalos M, Konno S, Eshghi M, Choudhury M (2010) Oxidative renal cell injury induced by calcium oxalate crystal and renoprotection with antioxidants: a possible role of oxidative stress in nephrolithiasis. J Endourol 24:339–345. CrossRefPubMedGoogle Scholar
  8. 8.
    Jonassen JA, Cao LC, Honeyman T, Scheid CR (2003) Mechanisms mediating oxalate-induced alterations in renal cell functions. Crit Rev Eukaryot Gene Expr 13:55–72CrossRefPubMedGoogle Scholar
  9. 9.
    Joshi S, Clapp WL, Wang W, Khan SR (2015) Osteogenic changes in kidneys of hyperoxaluric rats. Biochem Biophys Acta 1852:2000–2012. PubMedGoogle Scholar
  10. 10.
    Manissorn J, Fong-Ngern K, Peerapen P, Thongboonkerd V (2017) Systematic evaluation for effects of urine pH on calcium oxalate crystallization, crystal-cell adhesion and internalization into renal tubular cells. Sci Rep 7:1798. CrossRefPubMedGoogle Scholar
  11. 11.
    Wiessner JH, Hasegawa AT, Hung LY, Mandel GS, Mandel NS (2001) Mechanisms of calcium oxalate crystal attachment to injured renal collecting duct cells. Kidney Int 59:637–644. CrossRefPubMedGoogle Scholar
  12. 12.
    Niimi K, Yasui T, Okada A, Hirose Y, Kubota Y, Umemoto Y, Kawai N, Tozawa K, Kohri K (2014) Novel effect of the inhibitor of mitochondrial cyclophilin D activation, N-methyl-4-isoleucine cyclosporin, on renal calcium crystallization. Int J Urol 21:707–713. CrossRefPubMedGoogle Scholar
  13. 13.
    Sharma M, Sud A, Kaur T, Tandon C, Singla SK (2016) N-acetylcysteine with apocynin prevents hyperoxaluria-induced mitochondrial protein perturbations in nephrolithiasis. Free Radic Res 50:1032–1044. CrossRefPubMedGoogle Scholar
  14. 14.
    Sharma M, Kaur T, Singla SK (2016) Role of mitochondria and NADPH oxidase derived reactive oxygen species in hyperoxaluria induced nephrolithiasis: therapeutic intervention with combinatorial therapy of N-acetyl cysteine and Apocynin. Mitochondrion 27:15–24. CrossRefPubMedGoogle Scholar
  15. 15.
    Krols M, van Isterdael G, Asselbergh B, Kremer A, Lippens S, Timmerman V, Janssens S (2016) Mitochondria-associated membranes as hubs for neurodegeneration. Acta Neuropathol 131:505–523. CrossRefPubMedGoogle Scholar
  16. 16.
    Malhotra JD, Kaufman RJ (2011) ER stress and its functional link to mitochondria: role in cell survival and death. Cold Spring Harb Perspect Biol 3:a004424. PubMedGoogle Scholar
  17. 17.
    Guo B, Li Z (2014) Endoplasmic reticulum stress in hepatic steatosis and inflammatory bowel diseases. Front Genet 5:242. CrossRefPubMedGoogle Scholar
  18. 18.
    Jian L, Lu Y, Lu S, Lu C (2016) Chemical chaperone 4-phenylbutyric acid protects H9c2 cardiomyocytes from ischemia/reperfusion injury by attenuating endoplasmic reticulum stress-induced apoptosis. Mol Med Rep 13:4386–4392. CrossRefPubMedGoogle Scholar
  19. 19.
    Mohammed-Ali Z, Cruz GL, Dickhout JG (2015) Crosstalk between the unfolded protein response and NF-kappaB-mediated inflammation in the progression of chronic kidney disease. J Immunol Res 2015:428508. Google Scholar
  20. 20.
    Carlisle RE, Brimble E, Werner KE, Cruz GL, Ask K, Ingram AJ, Dickhout JG (2014) 4-Phenylbutyrate inhibits tunicamycin-induced acute kidney injury via CHOP/GADD153 repression. PLoS ONE 9:e84663. CrossRefPubMedGoogle Scholar
  21. 21.
    Zeng M, Sang W, Chen S, Chen R, Zhang H, Xue F, Li Z, Liu Y, Gong Y, Zhang H, Kong X (2017) 4-PBA inhibits LPS-induced inflammation through regulating ER stress and autophagy in acute lung injury models. Toxicol Lett 271:26–37. CrossRefPubMedGoogle Scholar
  22. 22.
    Guo Q, Xu L, Li H, Sun H, Liu J, Wu S, Zhou B (2017) Progranulin causes adipose insulin resistance via increased autophagy resulting from activated oxidative stress and endoplasmic reticulum stress. Lipids Health Dis 16:25. CrossRefPubMedGoogle Scholar
  23. 23.
    Hodgkinson A, Williams A (1972) An improved colorimetric procedure for urine oxalate. Clin Chim Acta 36:127–132CrossRefPubMedGoogle Scholar
  24. 24.
    Sharma M, Kaur T, Singla SK (2015) Protective effects of N-acetylcysteine against hyperoxaluria induced mitochondrial dysfunction in male wistar rats. Mol Cell Biochem 405:105–114. CrossRefPubMedGoogle Scholar
  25. 25.
    Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616CrossRefPubMedGoogle Scholar
  26. 26.
    Zahler W, Cleland W (1968) A specific and sensitive assay for disulfides. J Biol Chem 243:716–719PubMedGoogle Scholar
  27. 27.
    Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochem Biophys Acta 582:67–78. CrossRefPubMedGoogle Scholar
  28. 28.
    Aggarwal D, Gautam D, Sharma M, Singla SK (2016) Bergenin attenuates renal injury by reversing mitochondrial dysfunction in ethylene glycol induced hyperoxaluric rat model. Eur J Pharmacol 791:611–621. CrossRefPubMedGoogle Scholar
  29. 29.
    Umekawa T, Hatanaka Y, Kurita T, Khan SR (2004) Effect of angiotensin II receptor blockage on osteopontin expression and calcium oxalate crystal deposition in rat kidneys. J Am Soc Nephrol 15:635–644CrossRefPubMedGoogle Scholar
  30. 30.
    Sidhu H, Allison MJ, Chow JM, Clark A, Peck AB (2001) Rapid reversal of hyperoxaluria in a rat model after probiotic administration of Oxalobacter formigenes. J Urol 166:1487–1491CrossRefPubMedGoogle Scholar
  31. 31.
    Bijarnia RK, Bachtler M, Chandak PG, van Goor H, Pasch A (2015) Sodium thiosulfate ameliorates oxidative stress and preserves renal function in hyperoxaluric rats. PloS One 10:e0124881. CrossRefPubMedGoogle Scholar
  32. 32.
    Huang HS, Chen J, Chen CF, Ma MC (2006) Vitamin E attenuates crystal formation in rat kidneys: roles of renal tubular cell death and crystallization inhibitors. Kidney Int 70:699–710. CrossRefPubMedGoogle Scholar
  33. 33.
    Joshi S, Saylor BT, Wang W, Peck AB, Khan SR (2012) Apocynin-treatment reverses hyperoxaluria induced changes in NADPH oxidase system expression in rat kidneys: a transcriptional study. PloS ONE 7:e47738. CrossRefPubMedGoogle Scholar
  34. 34.
    Whittamore JM, Hatch M (2017) The role of intestinal oxalate transport in hyperoxaluria and the formation of kidney stones in animals and man. Urolithiasis 45:89–108. CrossRefPubMedGoogle Scholar
  35. 35.
    Joshi S, Wang W, Khan SR (2017) Transcriptional study of hyperoxaluria and calcium oxalate nephrolithiasis in male rats: inflammatory changes are mainly associated with crystal deposition. PLoS ONE 12:e0185009. CrossRefPubMedGoogle Scholar
  36. 36.
    Partovi N, Ebadzadeh MR, Fatemi SJ, Khaksari M (2017) Effect of fruit extract on renal stone formation and kidney injury in rats. Nat Prod Res. PubMedGoogle Scholar
  37. 37.
    Jonassen JA, Kohjimoto Y, Scheid CR, Schmidt M (2005) Oxalate toxicity in renal cells. Urol Res 33:329–339. CrossRefPubMedGoogle Scholar
  38. 38.
    Ferenbach DA, Bonventre JV (2016) Acute kidney injury and chronic kidney disease: from the laboratory to the clinic. Nephrol Ther 12(Suppl 1):S41–S48. CrossRefPubMedGoogle Scholar
  39. 39.
    Bonventre JV (2008) Kidney Injury Molecule-1 (KIM-1): a specific and sensitive biomarker of kidney injury. Scand J Clin Lab Invest Suppl 241:78–83. CrossRefPubMedGoogle Scholar
  40. 40.
    Vaidya VS, Waikar SS, Ferguson MA, Collings FB, Sunderland K, Gioules C, Bradwin G, Matsouaka R, Betensky RA, Curhan GC, Bonventre JV (2008) Urinary biomarkers for sensitive and specific detection of acute kidney injury in humans. Clin Transl Sci 1:200–208. CrossRefPubMedGoogle Scholar
  41. 41.
    Su H, Lei CT, Zhang C (2017) Interleukin-6 signaling pathway and its role in kidney disease: an update. Front Immunol 8:405. CrossRefPubMedGoogle Scholar
  42. 42.
    Imig JD, Ryan MJ (2013) Immune and inflammatory role in renal disease. Compr Physiol 3:957–976. PubMedGoogle Scholar
  43. 43.
    Umekawa T, Chegini N, Khan SR (2003) Increased expression of monocyte chemoattractant protein-1 (MCP-1) by renal epithelial cells in culture on exposure to calcium oxalate, phosphate and uric acid crystals. Nephrol Dial Transplant 18:664–669CrossRefPubMedGoogle Scholar
  44. 44.
    Umekawa T, Tsuji H, Uemura H, Khan SR (2009) Superoxide from NADPH oxidase as second messenger for the expression of osteopontin and monocyte chemoattractant protein-1 in renal epithelial cells exposed to calcium oxalate crystals. BJU Int 104:115–120. CrossRefPubMedGoogle Scholar
  45. 45.
    Bhardwaj R, Bhardwaj A, Tandon C, Dhawan DK, Bijarnia RK, Kaur T (2017) Implication of hyperoxaluria on osteopontin and ER stress mediated apoptosis in renal tissue of rats. Exp Mol Pathol 102:384–390. CrossRefPubMedGoogle Scholar
  46. 46.
    Chung AC, Lan HY (2011) Chemokines in renal injury. J Am Soc Nephrol 22:802–809. CrossRefPubMedGoogle Scholar
  47. 47.
    Aggarwal D, Kaushal R, Kaur T, Bijarnia RK, Puri S, Singla SK (2014) The most potent antilithiatic agent ameliorating renal dysfunction and oxidative stress from Bergenia ligulata rhizome. J Ethnopharmacol 158:85–93. CrossRefPubMedGoogle Scholar
  48. 48.
    Luo ZF, Feng B, Mu J, Qi W, Zeng W, Guo YH, Pang Q, Ye ZL, Liu L, Yuan FH (2010) Effects of 4-phenylbutyric acid on the process and development of diabetic nephropathy induced in rats by streptozotocin: regulation of endoplasmic reticulum stress-oxidative activation. Toxicol Appl Pharmacol 246:49–57. CrossRefPubMedGoogle Scholar
  49. 49.
    Bhardwaj R, Tandon C, Dhawan DK, Kaur T (2017) Effect of endoplasmic reticulum stress inhibition on hyperoxaluria-induced oxidative stress: influence on cellular ROS sources. World J Urol 35:1955–1965. CrossRefPubMedGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of BiochemistryPanjab University, ChandigarhChandigarhIndia

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