Angiotensin II induces alpha3(IV) collagen expression in cultured murine proximal tubular cells

Proc Assoc Am Physicians. 1999 Jul-Aug;111(4):357-64. doi: 10.1046/j.1525-1381.1999.99117.x.

Abstract

Angiotensin II (ANG II) induces cellular hypertrophy of cultured proximal tubular cells from various species. This hypertrophic response is associated with an increase in synthesis of basement membrane-associated collagen type IV. Previous investigations by our group have shown that ANG II stimulates mRNA and protein expression of the "classic" alpha1 and alpha2(IV) chains in cultured murine proximal tubular cells (murine cortical tubules [MCT cells]). Since it is clearer today that kidney basement membranes also contain heterotrimers of novel type IV collagens, the aim of the present study was to evaluate whether ANG II may influence the expression of alpha3 and alpha5(IV) collagen chains in MCT cells. A single dose of 10-8-10-6 M ANG II stimulated mRNA expression of alpha3(IV), but not of alpha5(IV), in MCT cells cultured in serum-free media. This response was mediated through AT1-receptors because losartan, but not an AT2-receptor antagonist, abolished the ANG II-induced expression of alpha3(IV) transcripts. Transient transfection of MCT cells with transforming growth factor-beta1 (TGF-beta1) antisense phosphorothioate-modified oligonucleotides partly abolished the ANG II-induced alpha3(IV) mRNA expression. Furthermore, Western blots of cellular lysates incubated with polyclonal antibodies generated against the recombinant collagen chains revealed that ANG II stimulated alpha3(IV) but not alpha5(IV) protein expression. This stimulation was partly prevented by co-incubation with a neutralizing anti-TGF-beta1-3 antibody. In summary, our data indicate that ANG II stimulates expression of the alpha3(IV) collagen chain in cultured MCT cells, due in part to TGF-beta1 activation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Angiotensin II / pharmacology*
  • Angiotensin Receptor Antagonists
  • Animals
  • Basement Membrane / drug effects
  • Basement Membrane / metabolism
  • Blotting, Northern
  • Blotting, Western
  • Collagen / biosynthesis*
  • Collagen / chemistry
  • Collagen / genetics
  • Culture Media, Serum-Free / pharmacology
  • Fibrosis
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression Regulation / drug effects*
  • Kidney Tubules, Proximal / drug effects*
  • Kidney Tubules, Proximal / metabolism
  • Losartan / pharmacology
  • Mice
  • Oligonucleotides, Antisense / pharmacology
  • Protein Conformation
  • Protein Isoforms / biosynthesis*
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Multimerization
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin / physiology
  • Transforming Growth Factor beta / biosynthesis
  • Transforming Growth Factor beta / genetics

Substances

  • Angiotensin Receptor Antagonists
  • Culture Media, Serum-Free
  • Oligonucleotides, Antisense
  • Protein Isoforms
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin
  • Transforming Growth Factor beta
  • Angiotensin II
  • Collagen
  • Losartan