title: Polyol effects on growth factors and MAPK signaling in rat retinal capillary cells.
abstract: OBJECTIVE
  Recent studies report that growth factor and signaling changes in rat lenses do not directly result from the presence of diabetes or sorbitol/galactitol (polyol) formation/accumulation, but from secondary osmotic changes associated with the aldose reductase (AR) catalyzed polyol formation. AR is also present in rat retinal pericyte and endothelial cells; however, significant polyol formation only occurs in pericytes and this does not appear to be linked to osmotic changes. The purpose of this study was to determine whether polyol formation and AR activity are similarly linked to growth factor and signaling changes in the rat capillary cells despite the apparent absence of osmotic stress.
  
  
  METHODS
  Conditionally immortalized rat retinal pericyte (TR-rPCT) and endothelial (TR-iBRB) cell lines were cultured on collagen type 1-coated dishes in the DMEM containing 5.5 mM glucose. After 24 h of initial culture, the medium was replaced with a serum-free medium containing 5.5, 25, or 50 mM glucose or galactose with/without the aldose reductase inhibitors (ARIs) AL1576 or tolrestat for periods of up to 48 h. Growth factors and transduction pathways were measured by Western blots using the antibodies against basic FGF, IGF-1, TGF-β, P-ERK1/2, P-SAPK/JNK, and P-Akt.
  
  
  RESULTS
  Sorbitol accumulation was only observed in pericytes, while galactitol was present in both pericytes and endothelial cells. Pericytes cultured in high glucose showed increased expression of the growth factors basic FGF, IGF-1, TGF-β, and signaling in P-Akt, P-ERK1/2, and P-SAPK/JNK compared with those cultured in 5.5 mM glucose and these expressions were normalized by the presence of ARIs. Similar results were observed with galactose media. In contrast, endothelial cells cultured in high glucose media showed neither growth factor or signaling changes. In galactose media, endothelial cells showed increased expression of basic FGF, IGF-1, TGF-β, P-ERK1/2, and P-SAPK/JNK, which were only partially reduced by ARIs.
  
  
  CONCLUSIONS
  Growth factor and MAPK signaling expression in pericytes are linked to the presence of polyols. Pericytes, which readily accumulate sorbitol/galactitol that is inhibited by ARIs, show expression changes similar to those observed in rat lenses. In contrast, endothelial cells only show partial expression changes that are linked to galactitol accumulation.
authors.affiliation: 1 Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center , Omaha, Nebraska.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Zifeng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Peter F
authors.initials: PF
authors.lastname: Kador
mesh_terms: D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D002196:Capillaries / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism; D005449:Fluorenes / Q000494:pharmacology; D004376:Galactitol / Q000378:metabolism*; D005690:Galactose / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D006827:Hydantoins / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology; D009281:Naphthalenes / Q000494:pharmacology; D020286:Pericytes / Q000378:metabolism; D051381:Rats; D055647:Rats, Transgenic; D012171:Retinal Vessels / Q000166:cytology / Q000378:metabolism*; D013012:Sorbitol / Q000378:metabolism*
chemical_list: D005449:Fluorenes; D006827:Hydantoins; D036341:Intercellular Signaling Peptides and Proteins; D009281:Naphthalenes; C052241:imirestat; C040550:tolrestat; D004376:Galactitol; D013012:Sorbitol; D000449:Aldehyde Reductase; D005947:Glucose; D005690:Galactose