Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation

Shambhu D Varma, Krish Chandrasekaran, Svitlana Kovtun Department of Ophthalmology and Visual Sciences, University of Maryland, Baltimore, MD, USA Purpose: Sulforaphane is a phytochemically derived organic isothiocyanate 1-isothiocyanato-4-methylsulfinyl-butane present naturally in crucifers, includ...

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Autores principales:	Varma SD, Chandrasekaran K, Kovtun S
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2013
Materias:	Ophthalmology RE1-994
Acceso en línea:	https://doaj.org/article/426873a3e3b04249b43679471d10e37c
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spelling	oai:doaj.org-article:426873a3e3b04249b43679471d10e37c2021-12-02T02:08:20ZSulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation1177-54671177-5483https://doaj.org/article/426873a3e3b04249b43679471d10e37c2013-10-01T00:00:00Zhttp://www.dovepress.com/sulforaphane-induced-transcription-of-thioredoxin-reductase-in-lens-po-a14778https://doaj.org/toc/1177-5467https://doaj.org/toc/1177-5483Shambhu D Varma, Krish Chandrasekaran, Svitlana Kovtun Department of Ophthalmology and Visual Sciences, University of Maryland, Baltimore, MD, USA Purpose: Sulforaphane is a phytochemically derived organic isothiocyanate 1-isothiocyanato-4-methylsulfinyl-butane present naturally in crucifers, including broccoli and cauliflower. Biochemically, it has been reported to induce the transcription of several antioxidant enzymes. Since such enzymes have been implicated in preventing cataract formation triggered by the intraocular generation of oxy-radical species, the purpose of this investigation was to examine whether it could induce the formation of antioxidant enzymes in the eye lens. Thioredoxin reductase (TrxR) was used as the target of such induction. Methods: Mice lenses were cultured for an overnight period of 17 hours in medium 199 fortified with 10% fetal calf serum. Incubation was conducted in the absence and presence of sulforaphane (5 µM). Subsequently, the lenses were homogenized in phosphate-buffered saline (PBS), followed by centrifugation. TrxR activity was determined in the supernatant by measuring the nicotinamide adenine dinucleotide phosphate (reduced) (NADPH)-dependent reduction of 5,5´-dithiobis-2-nitrobenzoic acid (DTNB). Non-specific reduction of DTNB was corrected for by conducting parallel determinations in the presence of aurothiomalate. The reduction of DTNB was followed spectrophotometrically at 410 nm. Results: The activity of TrxR in the lenses incubated with sulforaphane was found to be elevated to 18 times of that observed in lenses incubated without sulforaphane. It was also noticeably higher in the lenses incubated without sulforaphane than in the un-incubated fresh lenses. However, this increase was much lower than that observed for lenses incubated with sulforaphane. Conclusion: Sulforaphane has been found to enhance TrxR activity in the mouse lens in culture. In view of the protective effect of the antioxidant enzymes and certain nutrients against cataract formation, the findings suggest that it would, by virtue of its ability to enhance the activity of such enzymes, prevent the tissue against oxidative stress that leads to cataract formation. Additional studies with the activities of other antioxidant enzymes such as quinone oxidoreductase and the levels of Nrf2 are in progress. Keywords: oxidative stress, cataract, thioredoxin reductase, NADPHVarma SDChandrasekaran KKovtun SDove Medical PressarticleOphthalmologyRE1-994ENClinical Ophthalmology, Vol 2013, Iss default, Pp 2091-2098 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ophthalmology RE1-994
spellingShingle	Ophthalmology RE1-994 Varma SD Chandrasekaran K Kovtun S Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
description	Shambhu D Varma, Krish Chandrasekaran, Svitlana Kovtun Department of Ophthalmology and Visual Sciences, University of Maryland, Baltimore, MD, USA Purpose: Sulforaphane is a phytochemically derived organic isothiocyanate 1-isothiocyanato-4-methylsulfinyl-butane present naturally in crucifers, including broccoli and cauliflower. Biochemically, it has been reported to induce the transcription of several antioxidant enzymes. Since such enzymes have been implicated in preventing cataract formation triggered by the intraocular generation of oxy-radical species, the purpose of this investigation was to examine whether it could induce the formation of antioxidant enzymes in the eye lens. Thioredoxin reductase (TrxR) was used as the target of such induction. Methods: Mice lenses were cultured for an overnight period of 17 hours in medium 199 fortified with 10% fetal calf serum. Incubation was conducted in the absence and presence of sulforaphane (5 µM). Subsequently, the lenses were homogenized in phosphate-buffered saline (PBS), followed by centrifugation. TrxR activity was determined in the supernatant by measuring the nicotinamide adenine dinucleotide phosphate (reduced) (NADPH)-dependent reduction of 5,5´-dithiobis-2-nitrobenzoic acid (DTNB). Non-specific reduction of DTNB was corrected for by conducting parallel determinations in the presence of aurothiomalate. The reduction of DTNB was followed spectrophotometrically at 410 nm. Results: The activity of TrxR in the lenses incubated with sulforaphane was found to be elevated to 18 times of that observed in lenses incubated without sulforaphane. It was also noticeably higher in the lenses incubated without sulforaphane than in the un-incubated fresh lenses. However, this increase was much lower than that observed for lenses incubated with sulforaphane. Conclusion: Sulforaphane has been found to enhance TrxR activity in the mouse lens in culture. In view of the protective effect of the antioxidant enzymes and certain nutrients against cataract formation, the findings suggest that it would, by virtue of its ability to enhance the activity of such enzymes, prevent the tissue against oxidative stress that leads to cataract formation. Additional studies with the activities of other antioxidant enzymes such as quinone oxidoreductase and the levels of Nrf2 are in progress. Keywords: oxidative stress, cataract, thioredoxin reductase, NADPH
format	article
author	Varma SD Chandrasekaran K Kovtun S
author_facet	Varma SD Chandrasekaran K Kovtun S
author_sort	Varma SD
title	Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
title_short	Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
title_full	Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
title_fullStr	Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
title_full_unstemmed	Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
title_sort	sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation
publisher	Dove Medical Press
publishDate	2013
url	https://doaj.org/article/426873a3e3b04249b43679471d10e37c
work_keys_str_mv	AT varmasd sulforaphaneinducedtranscriptionofthioredoxinreductaseinlenspossiblesignificanceagainstcataractformation AT chandrasekarank sulforaphaneinducedtranscriptionofthioredoxinreductaseinlenspossiblesignificanceagainstcataractformation AT kovtuns sulforaphaneinducedtranscriptionofthioredoxinreductaseinlenspossiblesignificanceagainstcataractformation
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Sulforaphane-induced transcription of thioredoxin reductase in lens: possible significance against cataract formation

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