Localization of heparin and low-molecular-weight heparin in the rat kidney

 

 Buy Article Permissions and Reprints

Summary

Unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) are cleared, at least in part, by the kidneys through a poorly understood process. This study was undertaken to explore the mechanism of renal clearance of these drugs. Rats were given fluorescein-5-isothiocyanate (FITC)-labeled UFH or LMWH intravenously. At intervals after injection, rats were euthanized and the kidneys were harvested and subjected to immunohistochemical analysis and fluorescence microscopy. Both UFH and LMWH were localized to renal tubular cells and no immunoperoxidase staining or fluorescence was detected in glomeruli. Autoradiography demonstrated similar intracellular distribution of radio-labeled UFH suggesting that this phenomenon is independent of the method used to label heparin. Fluoresence in the tubules increased as a function of time after UFH injection, but reached a plateau after LMWH injection suggesting that the rate of renal tubular uptake depends on the molecular size of the heparin. When administered prior to FITC-labeled UFH or LMWH, probenecid, a renal organic anion inhibitor, decreased the renal tubular uptake of the heparins, whereas cimetidine, a renal organic cation inhibitor, had no effect. These findings suggest that renal excretion of UFH and LMWH primarily reflects tubular uptake via an organic anion transport mechanism.

• References

• 1 de Swart CAM, Nijmeyer B, Roelofs JMM. et al. Kinetics of intravenously administered heparin in normal humans. Blood 1982; 60: 1251-8.
  PubMedGoogle Scholar
• 2 Boneu B, Caranobe C, Cadroy Y. et al. Pharmacokinetic studies of standard unfractionated heparin, and low molecular weight heparins in the rabbit. Semin Thromb Hemost 1988; 14: 18-27.
  Article in Thieme ConnectPubMedGoogle Scholar
• 3 Barzu T, Molho P, Tobelem G. et al. Binding and endocytosis of heparin by human endothelial cells in culture. Biochim Biophys Acta 1985; 845: 196-203.
  CrossrefPubMedGoogle Scholar
• 4 Wells XE, Dawes J. Role of the liver and kidney in the desulphation of heparin in vivo. Thromb Haemost 1995; 74: 667-72.
  Article in Thieme ConnectPubMedGoogle Scholar
• 5 Barzu T, Van Rijn JLMC, Petitou M. et al. Heparin degradation in the endothelial cells. Thromb Res 1987; 47: 601-9.
  CrossrefPubMedGoogle Scholar
• 6 Fabian I, Bleiberg I, Aronson M. Increased uptake and desulphation of heparin by mouse macrophages in the presence of polycations. Biochim Biophys Acta 1978; 544: 69-76.
  CrossrefPubMedGoogle Scholar
• 7 Caranobe C, Barret A, Gabaig AM. et al. Disappearance of circulating anti-Xa activity after intravenous injection of standard heparin and of a low molecular weight heparin (CY216) in normal and nephrectomized rabbits. Thromb Res 1985; 40: 129-33.
  CrossrefPubMedGoogle Scholar
• 8 Palm M, Mattsson CH. Pharmacokinetics of heparin and low molecular weight heparin fragment (Fragmin) in rabbits with impaired renal or metabolic clearance. Thromb Haemost 1987; 58: 932-5.
  Article in Thieme ConnectPubMedGoogle Scholar
• 9 Young E, Cosmi B, Weitz J. et al. Comparison of the non-specific binding of unfractionated heparin and low molecular weight heparin (Enoxaparin) to plasma proteins. Thromb Haemost 1993; 70: 625-30.
  Article in Thieme ConnectPubMedGoogle Scholar
• 10 Young E, Wells P, Holloway S. et al. Ex-vivo and in-vitro evidence that low molecular weight heparins exhibit less binding to plasma proteins than unfractionated heparin. Thromb Haemost 1994; 71: 300-4.
  Article in Thieme ConnectPubMedGoogle Scholar
• 11 Weitz JI. Low-molecular-weight heparins. N Engl J Med 1997; 337: 688-98.
  CrossrefPubMedGoogle Scholar
• 12 Levine M, Gent M, Hirsh J. et al. A comparison of low-molecular-weight heparin administered primarily at home with unfractionated heparin administered in the hospital for proximal deep-vein thrombosis. N Engl J Med 1996; 334: 677-81.
  CrossrefPubMedGoogle Scholar
• 13 Koopman MMW, Prandoni P, Piovella F. et al. Treatment of venous thrombosis with intravenous unfractionated heparin administered in the hospital compared with subcutaneous low-molecular-weight heparin administered at home. N Engl J Med 1996; 334: 682-7.
  CrossrefPubMedGoogle Scholar
• 14 Shum DKY, Baylis C, Scott JE. A micropuncture and renal clearance study in the rat of the urinary excretion of heparin, chondroitin sulphate and metabolic breakdown products of connective tissue proteoglycans. Clin Sci 1984; 67: 205-12.
  CrossrefPubMedGoogle Scholar
• 15 Nagasawa K, Uchiyama H. Preparation and properties of biologically active fluorescent heparins. Biochim Biophys Acta 1978; 544: 430-40.
  CrossrefPubMedGoogle Scholar
• 16 Malsch R, Guerrini M, Torri G. et al. Synthesis of a N’-alkylamine anticoagulant active low-molecular-mass heparin for radioactive and fluorescent labeling. Anal Biochem 1994; 217: 255-64.
  CrossrefPubMedGoogle Scholar
• 17 Liaw PCY, Fredenburgh JC, Stafford AR. et al. Localization of the thrombin-binding domain on prothrombin fragment 2. J Biol Chem 1998; 273: 8932-9.
  CrossrefPubMedGoogle Scholar
• 18 Somogyi A. Renal transport of drugs: specificity and molecular mechanisms. Clin Exp Pharmacol Physiol 1996; 23: 986-9.
  CrossrefPubMedGoogle Scholar
• 19 Weiner IM, Roth L. Renal excretion of cimetidine. J Pharmacol Exp Ther 1981; 216: 516-20.
  PubMedGoogle Scholar
• 20 Beyer KJ, Russo HF, Tillson EK. et al. ‘Benemid’, p-(di-n-propylsulfamyl)-benzoic acid: its renal affinity and its elimination. Am J Physiol 1951; 166: 625-40.
  PubMedGoogle Scholar
• 21 de Miranda P, Good SS, Yarchoan R. et al. Alteration of zidovudine pharmacokinetics by probenecid in patients with AIDS or AIDSrelated complex. Clin Pharmacol Ther 1989; 46: 494-500.
  CrossrefPubMedGoogle Scholar
• 22 Lin JH, Chen IW, Deluna FA. et al. Renal handling of alendronate in rats. An uncharacterized renal transport system. Drug Metab Dispos 1992; 20: 608-13.
  PubMedGoogle Scholar
• 23 Foote EF, Halstenson CE. Effects of probenecid and cimetidine on renal disposition of ofloxacin in rats. Antimicrob Agents Chemother 1998; 42: 456-8.
  PubMedGoogle Scholar
• 24 Khorramian BA, Stivala SS. Small-angle X-ray scattering of high- and low-affinity heparin. Arch Biochem Biophys 1986; 247: 384-92.
  CrossrefPubMedGoogle Scholar
• 25 Kanwar YS, Farquhar MG. Presence of heparan sulfate in the glomerular basement membrane. Proc Natl Acad Sci USA 1979; 76: 1303-7.
  CrossrefPubMedGoogle Scholar
• 26 Sanchez G. Enhancement of heparin effect by probenecid. N Engl J Med 1975; 292: 48 (letter).
  PubMedGoogle Scholar
• 27 Sirak HD, McCleery RS, Artz CP. The effect of carinamide with heparin on the coagulation of human blood: a preliminary report. Surgery 1948; 24: 811.
  PubMedGoogle Scholar
• 28 Rodgers GM. Inappropriate use of enoxaparin in the treatment of deep-vein thrombosis. N Engl J Med 1999; 340: 62 (letter).
  PubMedGoogle Scholar
• 29 Rubin N, Lu S, Horn J. et al. The safety of low molecular weight heparin (LMWH) in renal failure: A case report of a large retroperitoneal hematoma as a complication of Lovenox use in a patient with renal failure. Blood 1998; 10: 125b (abstract).
  PubMedGoogle Scholar
• 30 Nagge J, Crowther M, Hirsh J. Is impaired renal function a contraindication to use of low-molecular-weight heparin?. Arch Intern Med 2002; 162: 2606-09.
  PubMedGoogle Scholar