The simple isocratic HPLC—UV method for the simultaneous determination of reduced and oxidized glutathione in animal tissue

Begić, Aida; Djurić, Ana; Gobeljić, Borko; Stevanović, Ivana; Lukić, Vera; Stanojević, Ivan; Ninković, Milica; Saso, Luciano; Vojvodić, Danilo; Djukić, Mirjana

doi:10.1556/1326.2017.29.1.5

Cited by 14 publications

(8 citation statements)

References 18 publications

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“…Liver tissues were homogenized in chilled 10 mM phosphate buffer solution (PBS) with optimal pH 7.4 and brain homogenate (1 g/10 ml, 75% aqueous methanol) was centrifuged and filtered. Both tissue homogenates were used to measure malondialdehyde (MDA) and glutathione (GSH) content using the high-performance liquid chromatography (HPLC) method [ 22 , 23 ]. The sample was suspended in acetonitrile, homogenized and centrifuged 3000 g for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Gallic acid and metformin co-administration reduce oxidative stress, apoptosis and inflammation via Fas/caspase-3 and NF-κB signaling pathways in thioacetamide-induced acute hepatic encephalopathy in rats

Mohamed

Hafez

2023

BMC Complement Med Ther

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Background Hepatic encephalopathy (HE) is a consequence of chronic or acute liver diseases. This study evaluates the combined effect of gallic acid (GA), and metformin (Met) on the liver and brain damage associated with HE. Methods Acute HE was induced by a single dose of thioacetamide (TAA) (300 mg/kg) as an I.P. injection. Treated groups received GA group (100 mg/kg/day, p.o), Met (200 mg/kg/day, p.o), or their combination for 25 consecutive days before TAA injection. Results The administration of TAA induced various biochemical and histopathological alterations. In contrast, treatment with GA either alone or combined with Met resulted in improved liver functions by the significant reduction in serum ALT, AST, and ALP activities, and ammonia levels. Inflammatory mediators; TNF-α, IL-6, and NFkβ levels were decreased by these treatments as well as apoptotic cascade via down-regulation of FAS and caspase-3 (CASP-3) expression in hepatic tissues. Furthermore, GA and Met either alone or combined protected the liver and brain tissues from damage by increased glutathione concentration while decreasing malondialdehyde. In addition, it was accompanied by the improvement of the brain neurotransmitter profile via the restoration of norepinephrine, dopamine, and serotonin levels. Based on our data, this is the first study to report a novel combined hepatoprotective and cognitive enhancing effect of GA and Met against TAA-induced acute liver and brain injury. Conclusion GA and Met combination resulted in a prominent improvement in HE complications, relative to monotherapy. Both agents potentiated the antioxidant, anti-inflammatory, and anti-apoptotic effects of each other.

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Section: Methodsmentioning

confidence: 99%

Gallic acid and metformin co-administration reduce oxidative stress, apoptosis and inflammation via Fas/caspase-3 and NF-κB signaling pathways in thioacetamide-induced acute hepatic encephalopathy in rats

Mohamed

Hafez

2023

BMC Complement Med Ther

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“…Mobile phase consisted of 100 mM sodium perchlorate solution (pH 2.8 adjusted with 0.1% ortho-phosphoric acid). Supernatant was used for chromatographic analysis16. The results were expressed in regard to protein status, as nmol GSH/mg proteins or nmol GSSG/mg proteins.…”

Section: Methodsmentioning

confidence: 99%

Disulfiram moderately restores impaired hepatic redox status of rats subchronically exposed to cadmium

Begić

Djurić

Ninković

et al. 2017

Journal of Enzyme Inhibition and Medicinal Chemistry

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Examination of cadmium (Cd) toxicity and disulfiram (DSF) effect on liver was focused on oxidative stress (OS), bioelements status, morphological and functional changes. Male Wistar rats were intraperitoneally treated with 1 mg CdCl2/kg BW/day; orally with 178.5 mg DSF/kg BW/day for 1, 3, 10 and 21 days; and co-exposed from 22nd to 42nd day. The co-exposure nearly restored previously suppressed total superoxide dismutase (SOD), catalase (CAT) and increased glutathione peroxidase (GPx) activities; increased previously reduced glutathione reductase (GR) and total glutathione-S-transferase (GST) activities; reduced previously increased superoxide anion radical (O2 ·−) and malondialdehyde (MDA) levels; increased zinc (Zn) and iron (Fe), and decreased copper (Cu) (yet above control value), while magnesium (Mg) was not affected; and decreased serum alanine aminotransferases (ALT) levels. Histopathological examination showed signs of inflammation process as previously demonstrated by exposure to Cd. Overall, we ascertained partial liver redox status improvement, compared with the formerly Cd-induced impact.

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“…Both GSH and GSSG were simultaneously measured using a modified high-performance liquid chromatography (HPLC-UV) method [17]. Isocratic chromatographic separation was carried out on a ZOR-BAX Eclipse AAA (4.6 x150 mm, 3.5 µm) analytical column (Agilent Technologies, USA.)…”

Section: Hplc-uv Determination Of Reduced and Oxidized Glutathionementioning

confidence: 99%

Disulfiram partially improves oxidative but not androgen status in rats exposed to cadmium

Pavlović

Đurić

Stevanović

et al. 2019

ARCH BIOL SCI BELGRA

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We investigated the effect of disulfiram (DSF) on reproductive toxicity induced by subchronic exposure to cadmium (Cd). We examined the redox status and systemic testosterone changes in the testes and plasma of Cd-treated male Wistar rats. Rats were treated with 1 mg CdCl 2 /kg body weight (bw)/day (intraperitoneal administration) for 42 days; in the second experimental group, rats were given 178.5 mg DSF/kg bw/day by oral gavage for 21 days; in the third group, after administration of Cd for 21 days, DSF treatment was introduced on day 22 and lasted until day 42, with continuous Cd intake. Each experimental group had a matching control: untreated rats, rats that received for 21 days olive oil, the solvent for DSF; rats that started with olive oil intake from days 22-42. Exposure of rats to DSF modulated the oxidative status in the testes; thus, coexposure increased the Cd-induced reduction in total superoxide dismutase (tSOD), catalase (CAT), glutathione reductase (GR) and total glutathione-S-transferase (tGST) activities, and lowered the Cd-increased superoxide anion radical (O 2 •-) and malondialdehyde (MDA) concentrations. DSF did not affect testosterone production diminished by Cd, as Leydig cells, once impaired by Cd, could not be reactivated by DSF.

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The simple isocratic HPLC—UV method for the simultaneous determination of reduced and oxidized glutathione in animal tissue

Cited by 14 publications

References 18 publications

Gallic acid and metformin co-administration reduce oxidative stress, apoptosis and inflammation via Fas/caspase-3 and NF-κB signaling pathways in thioacetamide-induced acute hepatic encephalopathy in rats

Gallic acid and metformin co-administration reduce oxidative stress, apoptosis and inflammation via Fas/caspase-3 and NF-κB signaling pathways in thioacetamide-induced acute hepatic encephalopathy in rats

Disulfiram moderately restores impaired hepatic redox status of rats subchronically exposed to cadmium

Disulfiram partially improves oxidative but not androgen status in rats exposed to cadmium

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