Chitosan Increases the Release of Renal Dipeptidase from Porcine Renal Proximal Tubule Cells

  • Hyun Joong, Yoon (College of Pharmacy, and Research Institute of Drug Development, Chonnam National University) ;
  • Kim, Young-Ho (Department of Biology, College of Natural Sciences, Chosun University) ;
  • Park, Sung-Wook (College of Pharmacy, and Research Institute of Drug Development, Chonnam National University) ;
  • Lee, Hwanghee-Blaise (Department of Biology, College of Natural Sciences, Chonnam National University) ;
  • Park, Haeng-Soon (College of Pharmacy, and Research Institute of Drug Development, Chonnam National University)
  • Published : 2003.12.01


Renal dipeptidase (RDPase, membrane dipeptidase, dehydropeptidase 1, EC has been widely studied since it was first purified from porcine kidney brush border membrane. It was reported that RDPase activity in urine samples of acute and chronic renal failure patients decreases. Nitric oxide (NO) is a highly reactive free radical involved in a number of physiological and pathological processes. NO is able to act in a dual mode, leading either to induction of apoptosis or to blunted execution of programmed cell death. NO inhibited the RDPase release from porcine renal proximal tubules, which could be blocked by L-NAME. Chitosan, the linear polymer of D-glucosamine in $\beta$(1\longrightarrow4) linkage, not only reversed the decreased RDPase release by NO but also increased NO production in the proximal tubule cells. The stimulatory effect of NO on RDPase release from proximal tubules in the presence of chitosan must be different from the previously proposed mechanism of RDPase release via NO signaling pathway. Chitosan stimulated the RDPase release in the proximal tubules and increased RDPase activity to 220% and 250% at 0.1% and 1%, respectively. RDPase release was decreased to about 40% in the injured proximal tubules and was recovered in proportion to the increase of chitosan. Chitosan may be useful in recovery of renal function from $HgCl_2$injury.


  1. Brewis IA, Turner AJ, and Hooper NM (1994) Activation of the glycosyl-phosphatidylinositol-anchored membrane dipeptidase upon release from pig kidney membranes by phospholipase C. Biochem J 303: 633-638
  2. Brewis IA, Ferguson MA, Mehlert A, Turner AJ, and Hooper NM (1995) Structures of the glycosyl-phosphatidylinositol anchors of porcine and human renal membrane dipeptidase. Comprehensive structural studies on the porcine anchor and interspecies comparison of the glycan core structures. J Biol Chem 270: 22946-22956
  3. Campbell BJ, Lin YC, Davis RV, and Ballew E (1966) The purification and properties of a particulate renal dipeptidase. Biochim Biophys Acta 118: 371-386
  4. Campbell BJ, Baker SF, Shukla SD, Forrester LJ, and Zahler WL (1990) Bioconversion of leukotriene D4 by lung dipeptidase. Biochim Biophys Acta 1042: 107-112
  5. Carina P, Ismael DB, Clelia MR, and Silvia GC (2003) Chitosan induces different L-arginine metabolic pathways in resting and inflammatory macrophages. Biochem Biophys Res Commun 304: 266-272
  6. Fukumura Y, Kera Y, Oshitani S, Ushijima Y, Kobayashi I, Liu Z, Watanabe T, Yamada R, Kikuchi H, Kawazu S, and Yabuuchi M (1999) Behaviour of urinary dipeptidase in patients with chronic renal failure. Ann Clin Biochem 36: 221-225
  7. Hassid A, Arabshahi H, Bourcier T, Dhaunsi GS, and Matthews C (1994) Nitric oxide selectively amplifies FGF-2-induced mitogenesis in primary rat aortic smooth muscle cells. Am J Physiol 267: H1040-1048
  8. Heneka MT, Loschmann PA, Gleichmann M, Weller M, Schulz JB, Wullner U, and Klockgether T (1998) Induction of nitric oxide synthase and nitric oxide-mediated apoptosis in neuronal PC12 cells after stimulation with tumor necrosis factor-${\alpha}$/ lipopolysaccharide. J Neurachem 71 : 88-94
  9. Hwang SM, Chen CY, Chen SS, and Chen JC (2000) Chitinous materials inhibit nitric oxide production by activated RAW 264.7 macrophages. Biochem Biophys Res Commun 271: 229-233
  10. Ishihara M, Fedarko NS, and Conrad HE (1987) Involvement of phosphatidylinositol and insulin in the coordinate regulation of proteoheparan sulfate metabolism and hepatocyte growth. J Biol Chem 262: 4708-4716
  11. Ito Y, Watanabe Y, Hirano K, Sugiura M, Sawaki S, and Ogiso T (1984) A fluorometric method for dipeptidase activity measurement in urine, using L-alanyl-L-alanine as substrate. J Biochem 96: 1-8
  12. Kera Y, Liu Z, Matsumoto T, Sorimachi Y, Nagasaki H, and Yamada RH (1999) Rat and human membrane dipeptidase: tissue distribution and developmental changes. Comp Biochem Physiol B 123: 53-58
  13. Klip A, Ramlal T, Douen AG, Burdett E, Young D, Cartee GD, and Holloszy JO (1988) Insulin-induced decrease in 5'-nucleotidase activity in skeletal muscle membranes. FEBS Lett 238: 419-423
  14. Kreisberg JI, Matthys E, and Venkatachalam MA (1983) Morphologic factors in acute renal failure. In: BM Brenner and JM Lazarus (ed), Acute Renal Failsure, WB Saunders Company, Philadelphia, pp 21-46
  15. Kropp H, Sundelof JG, Hajdu R, and Kahan FM (1982) Metabolism of thienamycin and related carbapenem antibiotics by the renal dipeptidase, dehydropeptidase. Antimicrob Agents Chemother 22: 62-70
  16. Kumar MNV (2000) A review of chitin and chitosan applications. React Funct Polym 46: 1-27
  17. Lakics V and Vogel SN (1998) Lipopolysaccharide and ceramide use divergent signaling pathways to induce cell death in murine macrophages. J Immunol 161: 2490-2500
  18. Llansola M, Monfort P, and Felipo V (2000) Inhibitors of phospholipase C prevent glutamate neurotoxicity in primary cultures of cerebellar neurons. J Pharmacol Exp Ther 292: 870-876
  19. Moncada S and Higgs EA (1993) The L-arginine-nitric oxide pathway. N Engl J Med 329: 2002-2012
  20. Moncada S, Palmer RM, and Higgs EA (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43: 109-142
  21. Movahedi S, Pang S, and Hooper NM (2000) Insulin stimulates the release of a subset of GPI-anchored proteins in a G-protein independent manner. Mol Membr Biol 17: 41-45
  22. Olsson LE, Wheeler MA, Sessa WC, and Weiss RM (1998) Bladder instillation and intraperitoneal injection of Escherichia coli lipopolysaccharide up-regulate cytokines and iNOS in rat urinary bladder. J Pharmacol Exp Ther 284: 1203-1208
  23. Pae HO, Seo WG, Kim NY, Oh GS, Kim GE, Kim YH, Kwak HJ, Yun YG, Jun CD, and Chung HT (2001) Induction of granulocytic differentiation in acute promyelocytic leukemia cells (HL-60) by water-soluble chitosan oligomer. Leuk Res 25: 339-346
  24. Park HS, Kim DH, Park SK, Kang SK, Burks M, Mullins JM, and Campbell BJ (1992) Biochem J Korea 25: 329-336
  25. Park SW, Yoon HJ, Lee HB, Hooper NM, and Park HS (2002) Nitric oxide inhibits the shedding of the glycosylphospha-tidylinositol-anchored dipeptidase from porcine renal proximal tubules. Biochem J 364: 211-218
  26. Pinsky DJ, Cai B, Tang X, Rodrigues C, Sciacca RR, and Cannon PJ (1995) The lethal effects of cytokine-induced nitric oxide on cardiac myocyte are blocked by nitric oxide synthase antagonism or transforming growth factor ${\beta}$. J Clin Invest 95: 677-685
  27. Romero G, Luttrell L, Rogol A, Zeller K, Hewlett E, and Lamer J (1988) Phosphatidylinositol-glycan anchors of membrane proteins: potential precursors of insulin mediators. Science 240: 509-511
  28. Rupprecht HD, Akagi Y, Keil A, and Hofer G (2000) Nitric oxide inhibits growth of glomerular mesangial cells: role of the transcription factor EGR-1. Kidney Int 57: 70-82
  29. Schmidt HH and Murad F (1991) Purification and characterization of a human NO synthase. Biochem Biophys Res Commun 181: 1372-1377
  30. Schmidt HH, Seifert R, and Bohme E (1989) Formation and release of nitric oxide from human neutrophils and HL-60 cells induced by a chemotactic peptide, platelet activating factor and leukotriene $B_4$. FEBS Lett 244: 357-360
  31. Smith SD, Wheeler MA, Foster HE Jr, and Weiss RM (1996) Urinary nitric oxide synthase activity and cyclic GMP levels are decreased with interstitial cystitis and increased with urinary tract infections. J Urol 155: 1432-1435
  32. Stuehr DJ and Nathan CF (1989) Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cell. J Exp Med 169: 1543-1555
  33. Taub M, Pollard JW, and Walker JM (1990) Methods of Molecular Biology. 5. Animal Cell Culture. Human Press, Clifton, pp 189-196
  34. Traylor LA, Proksch JW, Beanum VC, and Mayeux PR (1996) Nitric oxide generation by renal proximal tubules: role of nitric oxide in the cytotoxicity of lipid A. J Pharmacol Exp Ther 279: 91-96
  35. Wang D, Yu X, Cohen RA, and Brecher P (2000) Distinct effects of N-acetylcysteine and nitric oxide on angiotensin II-induced epidermal growth factor receptor phosphorylation and intracellular $Ca^{2+}$ levels. J Biol Chem 275: 12223-12230
  36. We JS, Kang BY, Lee JC, Lee HB, and Park HS (1997) Identification of urinary dipeptidase as the released form of renal dipeptidase. Kidney Blood Press Res 20: 411-415
  37. Welch CL and Campbell BJ (1980) Uptake of glycine from L-alanylglycine into renal brush border vesicles. J Membr Biol 54: 39-50
  38. Wheeler MA, Smith SD, Saito N, Foster HE Jr, and Weiss RM (1997) Effect of long-term oral L-arginine on the nitric oxide synthase pathway in the urine from patients with interstitial cystitis. J Urol 158: 2045-2050