Mechanism of action of hydrogen sulfide on cyclic AMP formation in rat retinal pigment epithelial cells

Ya Fatou Njie-Mbye, Madhura Kulkarni, Catherine A. Opere, Sunny E. Ohia

Research output: Contribution to journalArticle

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Abstract

Hydrogen sulfide (H 2S), a colorless gas with the pungent odor of rotten eggs has been reported to produce pharmacological actions in ocular and non-ocular tissues. We have evidence that H 2S, using sodium hydrosulfide (NaHS) and sodium sulfide (Na 2S) as donors can increase cyclic AMP (cAMP) production in neural retina. In the present study, we investigated the mechanism of action of H 2S on cyclic nucleotide production in rat retinal pigment epithelial cells (RPE-J). Cultured RPE-J cells were incubated for 30min in culture medium containing the cyclic nucleotide phosphodiesterase (PDE) inhibitor, IBMX (2mM). Cells were exposed to varying concentrations of NaHS, the H 2S substrate (l-cysteine), cyclooxygenase (COX) inhibitors or the diterpene activator of adenylate cyclase, forskolin in the presence or absence of H 2S biosynthetic enzymes or the ATP-sensitive potassium (K ATP) channel antagonist, glibenclamide. Following drug-treatment at different time intervals, cell homogenates were prepared for cAMP assay using a well established methodology. In RPE-J cells, NaHS (10nM-1μM) produced a time-dependent increase in cAMP concentrations over basal levels which reached a maximum at 20min. At this time point, both NaHS (1nM-100μM) and l-cysteine (1nM-10μM) produced a concentration-dependent significant (pATP channel antagonist, glibenclamide (100μM) caused inhibition of NaHS induced-increase of cAMP formation in RPE-J cells. We conclude that, H 2S (using H 2S donor and substrate) can increase cAMP production in RPE-J cells, and removal of the apparent inhibitory effect of prostaglandins unmasks an excitatory activity of H 2S on cAMP. Effects elicited by the H 2S substrate on cAMP formation are dependent on biosynthesis of H 2S catalyzed by the biosynthetic enzymes, CBS and CSE. In addition to the adenylyl cylcase pathway, K ATP channels are involved in mediating the observed effects of the H 2S on cAMP production.

Original languageEnglish
Pages (from-to)16-22
Number of pages7
JournalExperimental Eye Research
Volume98
Issue number1
DOIs
StatePublished - May 2012

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Hydrogen Sulfide
Retinal Pigments
Cyclic AMP
Epithelial Cells
Glyburide
Cyclic Nucleotides
Cysteine
Adenosine Triphosphate
1-Methyl-3-isobutylxanthine
KATP Channels
Phosphodiesterase Inhibitors
Cyclooxygenase Inhibitors
Diterpenes
Colforsin
Enzymes
Adenylyl Cyclases
Eggs
Prostaglandins
Culture Media
Retina

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Mechanism of action of hydrogen sulfide on cyclic AMP formation in rat retinal pigment epithelial cells. / Njie-Mbye, Ya Fatou; Kulkarni, Madhura; Opere, Catherine A.; Ohia, Sunny E.

In: Experimental Eye Research, Vol. 98, No. 1, 05.2012, p. 16-22.

Research output: Contribution to journalArticle

Njie-Mbye, Ya Fatou ; Kulkarni, Madhura ; Opere, Catherine A. ; Ohia, Sunny E. / Mechanism of action of hydrogen sulfide on cyclic AMP formation in rat retinal pigment epithelial cells. In: Experimental Eye Research. 2012 ; Vol. 98, No. 1. pp. 16-22.
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