L-buthionine sulfoximine-induced loss of glutathione does not elicit PGH synthase inactivation in cultured bovine lens epithelial cells

J Ocul Pharmacol. 1990 Spring;6(1):43-9. doi: 10.1089/jop.1990.6.43.

Abstract

Previous studies from this laboratory have indicated that exposure of cultured bovine lens epithelial cells (BLECs) to minimal essential medium (MEM) containing 40 mM galactose (Gal) for as short a duration as 20 h results in a reduction of microsomal prostaglandin biosynthesis as demonstrated by a decrease in PGH synthase activity (Invest. Ophthalmol. Vis. Sci. 29:1452-1460, 1988). The present study shows that upon brief exposure of BLECs to Gal, the cellular content of glutathione (GSH) decreases as galactitol increases. Studies were therefore undertaken to establish whether a positive correlation existed between polyol accumulation, GSH content and the activity of PGH synthase (an enzyme known to auto-oxidize) utilizing BLECs exposed to hypergalactosemic conditions. The inhibitor of glutathione biosynthesis, L-buthionine sulfoximine (L-BSO) was used in order to lower the intracellular pool of GSH in MEM-incubated cells to a level below that routinely observed in Gal-incubated cells, under conditions whereby no galactitol accumulation had occurred. The galactitol content was 92 nmol/micrograms PO4 in Gal-incubated cells after a 20 h exposure period; no detectable level of galactitol was observed in BLECs maintained in galactose-free MEM. L-(BSO) (1 mM) was co-administered to BLECs maintained in either MEM or Gal for 20 h. The cellular content of GSH was 1.70 +/- .02 micrograms GSH/micrograms PO4 in MEM alone and 0.540 +/- .02 micrograms GSH/micrograms PO4 in MEM + BSO. Furthermore, the GSH content in BLECs after 20 h of exposure to Gal was 0.960 +/- .01 microgram GSH/microgram PO4 but decreased to 0.110 +/- .01 microgram GSH/microgram PO4 in Gal + BSO.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

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

MeSH terms

  • Aldehyde Reductase / metabolism
  • Animals
  • Antimetabolites / pharmacology*
  • Buthionine Sulfoximine
  • Cattle
  • Cells, Cultured
  • Culture Media
  • Enzyme Activation
  • Epithelial Cells
  • Galactose / pharmacology
  • Glutathione / biosynthesis*
  • Intracellular Fluid / metabolism
  • Lens Capsule, Crystalline / enzymology*
  • Lens Capsule, Crystalline / metabolism
  • Lens, Crystalline / enzymology*
  • Methionine Sulfoximine / analogs & derivatives*
  • Methionine Sulfoximine / pharmacology
  • Polymers / metabolism
  • Prostaglandin-Endoperoxide Synthases / metabolism*

Substances

  • Antimetabolites
  • Culture Media
  • Polymers
  • polyol
  • Methionine Sulfoximine
  • Buthionine Sulfoximine
  • Aldehyde Reductase
  • Prostaglandin-Endoperoxide Synthases
  • Glutathione
  • Galactose