A fluorometric method for determination of oxidized and reduced glutathione in tissues

Hissin, Paul J.; Hilf, Russell

doi:10.1016/0003-2697(76)90326-2

Cited by 3,808 publications

(1,572 citation statements)

References 17 publications

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“…Liver (250 mg) was homogenized in 3.75 ml of the phosphate - EDTA buffer (pH-8.0), 1 ml of 25% H 3 PO 4 and centrifuged at 4°C at 100,000 g for 30 min and the supernatant obtained was used for the assay of reduced (GSH) and oxidised glutathione (GSSG) as per Hissin and Hilf [38]. …”

Section: Methodsmentioning

confidence: 99%

Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver

et al. 2010

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BackgroundCarbon tetra chloride (CCl4), an industrial solvent, is a hepatotoxic agent and it is the well established animal model for free radical-induced liver injury. The present investigation was carried out to establish the protective effect of natansnin, a novel dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver.ResultsCCl4 significantly increased the levels of lipid peroxides, oxidized glutathione and decreased the levels of reduced glutathione, SOD and CAT. CCl4 induce marked histopathological changes and increase in the levels of apoptotic proteins. CCl4 treatment significantly increased the levels of apoptotic proteins such as caspases-3, PARP, Bax, Bid and cytochrome C and also increased the levels of inflammatory mediators iNos and Cox-2. Natansnin treatment significantly decreased the levels of CCl4 induced apoptotic proteins and inflammatory mediators. Further natansinin treatment significantly inhibited the CCl4 induced apoptosis which was evident form the reduced TUNEL positive cells.ConclusionsIn conclusion, our study demonstrated the protective effect of natansnin against CCl4 induced oxidative stress and cellular degeneration in rat liver tissue. This protective effect of natansnin can be correlated to its direct antioxidant effect.

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Section: Methodsmentioning

confidence: 99%

Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver

et al. 2010

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“…Determination of GSH was performed by the method of Hissin and Hilf [53]. The reaction mixture containing 0.1 M sodium phosphate buffer (pH-8.0), 5.0 mM EDTA, 10 μl Ophthaldehyde (1.0 mg/ml) and 10 μl of sample.…”

Section: Estimation Of Reduced Glutathione (Gsh)mentioning

confidence: 99%

“…The estimation of GSSG was performed by the method of Hissin and Hilf [53]. The samples were incubated first with 0.04 M N-ethyleimide (NEM) for 30 min to interact with GSH present in sample.…”

Section: Estimation Of Oxidized Glutathione (Gssg)mentioning

confidence: 99%

In vivo administration of D609 leads to protection of subsequently isolated gerbil brain mitochondria subjected to in vitro oxidative stress induced by amyloid beta-peptide and other oxidative stressors: Relevance to Alzheimer’s disease and other oxidative stress-related neurodegenerative disorders

Ansari

Joshi

Huang

et al. 2006

Free Radical Biology and Medicine

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Tricyclodecan-9-yl-xanthogenate (D609) has in vivo and in vitro antioxidant properties. D609 mimics glutathione (GSH), has a free thiol group, which upon oxidation forms a disulfide. The resulting dixanthate is a substrate for glutathione reductase, regenerating D609. Recent studies have also shown that D609 protects brain in vivo and neuronal cultures in vitro against the potential Alzheimer's disease (AD) causative factor, Aβ (1-42)-induced oxidative stress and cytotoxicity. Mitochondria are important organelles with both pro-and anti-apoptotic factor proteins. The present study was undertaken to test the hypothesis that i.p. injection of D609 would provide neuroprotection against free radical-induced, mitochondria-mediated apoptosis in vitro. Brain mitochondria were isolated from gerbils 1 h post injection intraperitonially (i.p.) with D609 and subsequently treated in vitro with the oxidants Fe 2+ /H 2 O 2 (hydroxyl free radicals), 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH, alkoxyl and peroxyl free radicals) and AD-relevant amyloid β-peptide 1-42 [Aβ (1-42)]. Brain mitochondria isolated from the gerbils previously injected i.p. with D609 and subjected to these oxidative stress inducers, in vitro, showed significant reduction in levels of protein carbonyls, protein-bound hydroxynonenal (HNE) [a lipid peroxidation product], 3-nitrotyrosine (3-NT), and cytochrome-c release compared to oxidant-treated brain mitochondria isolated from salineinjected gerbils. D609 treatment significantly maintains the GSH/GSSG ratio in oxidant-treated mitochondria. Increased activity of glutathione-S-transferase (GST), glutathione peroxidase (GPx) and glutathione reductase (GR) in brain isolated from D609-injected gerbils is consistent with the notion that D609 acts like GSH. These anti-apoptotic findings are discussed with reference to the potential use of this brain-accessible glutathione mimetic in the treatment of oxidative stress-related neurodegenerative disorders, including AD.

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“…GSH and GSSG levels were measured by method of Hissin and Hilf, 1976). To 10 ll homogenate, 2.8 ml of the phosphated-EDTA buffer (pH 8.0), and 100 ll Ophthalaldehyde solution (2 mg ml À1 in methanol) were added.…”

Section: Gsh and Gssg Measurementsmentioning

confidence: 99%

Effects of hexachlorobenzene on antioxidant status of liver and brain of common carp (Cyprinus carpio)

2006

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A fluorometric method for determination of oxidized and reduced glutathione in tissues

Cited by 3,808 publications

References 17 publications

Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver

Protective efficacy of natansnin, a dibenzoyl glycoside from Salvinia natans against CCl4 induced oxidative stress and cellular degeneration in rat liver

Effects of hexachlorobenzene on antioxidant status of liver and brain of common carp (Cyprinus carpio)

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