Polyol formation in cell lines of rat retinal capillary pericytes and endothelial cells (TR-rPCT and TR-iBRB)

Peter F Kador, James Randazzo, Karen Blessing, Jun Makita, Peng Zhang, Kuang Yu, Ken Ichi Hosoya, T. Terasaki

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Purpose: The two most widely investigated animal models for diabetic retinopathy (DR) are the rat and dog. In dogs, aldose reductase (AR) is present only in retinal capillary pericytes and their destruction has been linked to polyol accumulation and resulting apoptosis. Since both rat capillary pericytes and endothelial cells have been reported to contain AR, the role of polyol pathway activity in capillary cell destruction has been investigated in rat retinal capillary pericyte (TR-rPCT) and endothelial (TR-iBRB) cells. Methods: TR-rPCT and TR-iBRB cell lines were recloned and their identities were reconfirmed by characteristic immunostaining. Cells were cultured up to 72 h in media containing 50 mM glucose or galactose with/without the AR inhibitors or a sorbitol dehydrogenase inhibitor (SDI) or with 30 mM 3-fluoro-3-deoxyglucose. Polyol levels were determined by HPLC or 19F-NMR. Apoptosis was detected with TUNEL/DAPI staining. Results: Smooth muscle actin is present only in pericytes while only endothelial cells stain for von Willebrand factor and accumulate acetylated low-density lipoprotein. AR is present in both cells but AR levels are lower in endothelial cells. Aldehyde reductase is also present in both cells. Cells cultured in 50 mM glucose or galactose show significant polyol accumulation in pericytes but endothelial cells show little accumulation of galactitol and no accumulation of sorbitol. Sorbitol accumulation in pericytes resulted in increased cellular permeability and increased TUNEL staining, which was reduced by AR inhibition. Conclusions: Although both rat retinal pericytes and endothelial cells contain AR, sorbitol accumulation and TUNEL staining primarily occur in pericytes and are inhibited by AR inhibitors.

Original languageEnglish (US)
Pages (from-to)299-307
Number of pages9
JournalJournal of Ocular Pharmacology and Therapeutics
Volume25
Issue number4
DOIs
StatePublished - Aug 1 2009

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Aldehyde Reductase
Pericytes
Endothelial Cells
Cell Line
Sorbitol
In Situ Nick-End Labeling
Staining and Labeling
Galactose
Cultured Cells
Galactitol
L-Iditol 2-Dehydrogenase
Dogs
Apoptosis
polyol
Glucose
von Willebrand Factor
Diabetic Retinopathy
Smooth Muscle
Actins
Permeability

ASJC Scopus subject areas

  • Ophthalmology
  • Pharmacology
  • Pharmacology (medical)

Cite this

Polyol formation in cell lines of rat retinal capillary pericytes and endothelial cells (TR-rPCT and TR-iBRB). / Kador, Peter F; Randazzo, James; Blessing, Karen; Makita, Jun; Zhang, Peng; Yu, Kuang; Hosoya, Ken Ichi; Terasaki, T.

In: Journal of Ocular Pharmacology and Therapeutics, Vol. 25, No. 4, 01.08.2009, p. 299-307.

Research output: Contribution to journalArticle

Kador, Peter F ; Randazzo, James ; Blessing, Karen ; Makita, Jun ; Zhang, Peng ; Yu, Kuang ; Hosoya, Ken Ichi ; Terasaki, T. / Polyol formation in cell lines of rat retinal capillary pericytes and endothelial cells (TR-rPCT and TR-iBRB). In: Journal of Ocular Pharmacology and Therapeutics. 2009 ; Vol. 25, No. 4. pp. 299-307.
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AU - Zhang, Peng

AU - Yu, Kuang

AU - Hosoya, Ken Ichi

AU - Terasaki, T.

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