Interleukin-1-induced alterations in glomerular proteoglycans: Biochemical and tissue autoradiographic aspects

Naoki Kashihara, Yuzo Watanabe, Hirofumi Makino, Yashpal S. Kanwar

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The effect of interleukin-1α (IL-1) on the synthesis of glomerular basement membrane heparan sulfateproteoglycan (HS-PG) was investigated. An ex vivo recirculating organ perfusion system was used. Kidneys were perfused with a medium containing (35S) sulfate and IL-1 (0.625 to 6.25 ng/mL). After radiolabeling was performed, a small cortical piece was saved for tissue autoradiography; the remaining kidney and perfusion medium were used for biochemical studies. Renal cortices were dissected out, and glomeruli were isolated; the PG were extracted and characterized. With exposure to IL-1 (5 ng/mL), an approximately twofold increase of the radioactivity in the glomerular fraction was noted. The increase was unaffected by indomethacin treatment. By Sepharose CL-6B chromatography, a single peak of radioactivity with Kav = 0.25 (macromolecular form of PG) was observed in the control group. The IL-treated group had two peaks of radioactivity with Kav = 0.25 and 0.45: the first peak contained PG, and the second peak consisted of free glycosaminoglycan (GAG) chains. Elution profiles of hydrolyzed tissue GAG chains were similar. No change in the ratio of chondroitin to heparan sulfate was observed. By DEAE-Sephacel chromatography, the glomerular PG/ GAG of the IL-treated group eluted at a relatively lower salt concentration, suggesting a change in the charge density characteristics. No differences in the elution profiles of media PG/GAG were observed. (35S)methionine labeling of proteins showed no significant increase of the total radioincorporation in the IL-treated group, Immunoprecipitation studies revealed an approximately 88% increase in the de novo synthesis of PG. Tissue autoradiography revealed an approximately twofold increase of (35S) sulfate radioactivity over the glomerular mesangium, epithelium, and basement membranes. These results indicate that IL-1 enhances the synthesis of the macromolecular form of PG and the generation of free chains. Conceivably, such alterations may lead to defective macromolecular interactions among various components of the glomerular basement membrane and compromise the integrity of the ultrafiltration unit of the glomerulus.

Original languageEnglish
Pages (from-to)203-213
Number of pages11
JournalJournal of the American Society of Nephrology
Volume3
Issue number2
Publication statusPublished - Aug 1 1992
Externally publishedYes

Fingerprint

Proteoglycans
Glycosaminoglycans
Interleukin-1
Radioactivity
Glomerular Basement Membrane
Autoradiography
Kidney
Sulfates
Chromatography
Perfusion
Glomerular Mesangium
Chondroitin
Heparitin Sulfate
Interleukin-5
Ultrafiltration
Immunoprecipitation
Basement Membrane
Indomethacin
Methionine
Epithelium

Keywords

  • Extracellular matrix
  • Glomerular cells
  • Interleukin
  • Proteoglycans

ASJC Scopus subject areas

  • Nephrology

Cite this

Interleukin-1-induced alterations in glomerular proteoglycans : Biochemical and tissue autoradiographic aspects. / Kashihara, Naoki; Watanabe, Yuzo; Makino, Hirofumi; Kanwar, Yashpal S.

In: Journal of the American Society of Nephrology, Vol. 3, No. 2, 01.08.1992, p. 203-213.

Research output: Contribution to journalArticle

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N2 - The effect of interleukin-1α (IL-1) on the synthesis of glomerular basement membrane heparan sulfateproteoglycan (HS-PG) was investigated. An ex vivo recirculating organ perfusion system was used. Kidneys were perfused with a medium containing (35S) sulfate and IL-1 (0.625 to 6.25 ng/mL). After radiolabeling was performed, a small cortical piece was saved for tissue autoradiography; the remaining kidney and perfusion medium were used for biochemical studies. Renal cortices were dissected out, and glomeruli were isolated; the PG were extracted and characterized. With exposure to IL-1 (5 ng/mL), an approximately twofold increase of the radioactivity in the glomerular fraction was noted. The increase was unaffected by indomethacin treatment. By Sepharose CL-6B chromatography, a single peak of radioactivity with Kav = 0.25 (macromolecular form of PG) was observed in the control group. The IL-treated group had two peaks of radioactivity with Kav = 0.25 and 0.45: the first peak contained PG, and the second peak consisted of free glycosaminoglycan (GAG) chains. Elution profiles of hydrolyzed tissue GAG chains were similar. No change in the ratio of chondroitin to heparan sulfate was observed. By DEAE-Sephacel chromatography, the glomerular PG/ GAG of the IL-treated group eluted at a relatively lower salt concentration, suggesting a change in the charge density characteristics. No differences in the elution profiles of media PG/GAG were observed. (35S)methionine labeling of proteins showed no significant increase of the total radioincorporation in the IL-treated group, Immunoprecipitation studies revealed an approximately 88% increase in the de novo synthesis of PG. Tissue autoradiography revealed an approximately twofold increase of (35S) sulfate radioactivity over the glomerular mesangium, epithelium, and basement membranes. These results indicate that IL-1 enhances the synthesis of the macromolecular form of PG and the generation of free chains. Conceivably, such alterations may lead to defective macromolecular interactions among various components of the glomerular basement membrane and compromise the integrity of the ultrafiltration unit of the glomerulus.

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