An Immunohistochemical Investigation of Renal Phospholipidosis and Toxicity in Rats

Int J Toxicol. 2017 Sep/Oct;36(5):386-394. doi: 10.1177/1091581817726040. Epub 2017 Aug 18.

Abstract

Immunohistochemical staining for the lysosome-associated membrane protein 2 (LAMP-2) has been proposed previously as an alternative to electron microscopy to identify hepatic phospholipidosis. This study used LAMP-2 immunohistochemistry (IHC) to diagnose phospholipidosis in rats exhibiting renal tubular injury. Rats were administered toreforant, a histamine H4 receptor antagonist by oral gavage at a dose of 3, 10, or 100 mg/kg/d for 6 months. Hematoxylin and eosin staining revealed renal tubular epithelial cell vacuolation, hypertrophy, degeneration, and luminal dilation in the 100 mg/kg/d group animals. Renal tubular injury was confirmed using kidney injury marker 1 (KIM-1) IHC. The involvement of phosopholipidosis in the renal injury was investigated by LAMP-2. Adipophilin IHC was included to differentiate phospholipidosis from lipidosis. Increased LAMP-2 staining was observed in the 100 mg/kg/d group animals when compared to vehicle group animals. Lysosome-associated membrane protein-2 staining was most prominent in the outer stripe of the outer medulla where KIM-1 staining was also most prominent. By contrast, adipophilin staining was not increased. Phospholipidosis was also confirmed by electron microscopy. These data support the use of LAMP-2 IHC as a diagnostic tool and suggest an association between phospholipidosis and the renal tubular injury caused by toreforant.

Keywords: KIM-1; LAMP-2; immunohistochemistry; kidney; phospholipidosis.

MeSH terms

  • Acute Kidney Injury
  • Animals
  • Cell Adhesion Molecules / metabolism
  • Female
  • Histamine Antagonists / toxicity*
  • Immunohistochemistry
  • Kidney / drug effects*
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney / ultrastructure
  • Kidney Diseases / chemically induced
  • Kidney Diseases / diagnosis*
  • Kidney Diseases / metabolism
  • Kidney Diseases / pathology
  • Lipidoses / chemically induced
  • Lipidoses / diagnosis*
  • Lipidoses / metabolism
  • Lipidoses / pathology
  • Lysosomal-Associated Membrane Protein 2 / analysis
  • Lysosomal-Associated Membrane Protein 2 / metabolism*
  • Male
  • Microscopy, Electron, Transmission
  • Perilipin-2 / analysis
  • Perilipin-2 / metabolism
  • Phospholipids / metabolism*
  • Rats, Sprague-Dawley
  • Receptors, Histamine H4 / antagonists & inhibitors*

Substances

  • Cell Adhesion Molecules
  • Havcr1protein, rat
  • Histamine Antagonists
  • Lysosomal-Associated Membrane Protein 2
  • Perilipin-2
  • Phospholipids
  • Plin2 protein, rat
  • Receptors, Histamine H4