scholarly journals Experience of measuring glutathione peroxidase activity in surgically induced endometrial-like lesions in rats

2021 ◽  
Vol 70 (2) ◽  
pp. 55-61
Author(s):  
Aleksey V. Razygraev ◽  
Elena V. Baziyan ◽  
Lyudmila S. Polyanskikh ◽  
Mariya A. Petrosyan

BACKGROUND: Endometriosis is known to be linked with altered activities of antioxidant enzymes and with their gene polymorphisms. Progestins are known to induce glutathione peroxidase activity in the endometrium and promote reduction of endometrial lesions. It could be useful to estimate the correlation between the activity of glutathione peroxidase within endometrial lesions and their degree of reduction. AIM: The present study was aimed at estimating glutathione peroxidase activity in surgically induced endometrial-like lesions of different degree of reduction in rat model of endometriosis. MATERIALS AND METHODS: The method for determining glutathione peroxidase activity using hydrogen peroxide as a substrate and 5,5-dithiobis(2-nitrobenzoic acid) for estimation of residual reduced glutathione was applied for quantitative analysis of the enzyme activity in endometriotic foci, surgically induced in female Wistar rats. An assay of glutathione peroxidase activity in tissue homogenates was performed at 37C in a reaction medium containing Tris-HCl buffer supplemented with tetrasodium ethylenediaminetetraacetate and sodium azide (pH 8.5) in the presence of 0.55 mM reduced glutathione and 0.192 mM hydrogen peroxide. Before adding trichloroacetic acid, 40-second incubation was used. The correlation between the specific activity of the enzyme and protein amount in endometriotic foci was estimated. RESULTS: In a rat model of endometriosis, there was a high, well-determined glutathione peroxidase activity in endometriotic foci. For the same endometriotic tissue sample, the enzymatic activity was proportional to the amount of protein in the reaction mixture. The range of specific glutathione peroxidase activity was 2.436.45 micromoles of consumed glutathione per minute per milligram of protein (n = 7). In most reduced endometriotic foci (with the minimum amount of endometriotic tissue), the highest specific activity of glutathione peroxidase was found (the Spearmans rho of 0.93 with p = 0.0067). CONCLUSIONS: The method for determining glutathione peroxidase activity using hydrogen peroxide and 5,5-dithiobis(2-nitrobenzoic acid) is convenient for working with the endometriotic tissue in a rat model of endometriosis. We can accept, with p 0.01, that weight of endometriotic foci is negatively linked with specific glutathione peroxidase activity within their tissue. The results are analogous to the previously obtained data on catalase activity and suggest the involvement of both antioxidant enzymes in reduction of endometrial lesions.

2006 ◽  
Vol 1091 (1) ◽  
pp. 10-16 ◽  
Author(s):  
MARIA CRISTINA ALBERTINI ◽  
FLAVIA RADOGNA ◽  
AUGUSTO ACCORSI ◽  
FRANCESCO UGUCCIONI ◽  
LAURA PATERNOSTER ◽  
...  

2004 ◽  
Vol 52 (3) ◽  
pp. 369-378 ◽  
Author(s):  
Katalin Németh ◽  
M. Mézes ◽  
T. Gaál ◽  
Á. Bartos ◽  
K. Balogh ◽  
...  

The effect of supplementary methionine and fats of different saturation levels on the glutathione redox system of growing broiler cockerels was studied. The diet of three groups of chicks was supplemented with corn germ oil, beef tallow and fish oil at the levels of 30 g/kg and 50 g/kg of feed, respectively. The diet of further three groups was supplemented with methionine (5 g/kg of feed) in addition to the different fat sources. Control chicks were fed with a compound feed without methionine and fat supplementation. Reduced glutathione (GSH) and glutathione disulphide (GSSG) content as well as glutathione peroxidase activity in the liver were determined and GSH/GSSG ratio was calculated at day old and then at one and three weeks of age. Our results indicate that supplementary methionine stimulates both the synthesis of the glutathione redox system and glutathione peroxidase activity in growing chickens in the first period of postnatal life, when the risk of lipid peroxidation is high due to feeding unsaturated fats in the diet.


2010 ◽  
Vol 79 (1) ◽  
pp. 27-31 ◽  
Author(s):  
Mária Weber ◽  
Krisztián Balogh ◽  
Judit Fodor ◽  
Márta Erdélyi ◽  
Zsolt Ancsin ◽  
...  

The effect of T-2 and HT-2 toxin using different doses in the starter (0-21 days: 1.04 mg T-2 toxin and 0.49 mg HT-2 toxin kg-1feed), and finisher diets (22-39 days: 0.12 mg T-2 toxin and 0.02 mg HT-2 toxin kg-1feed) was investigated in broiler chickens. Birds were divided into two groups fed with control and T-2 and HT-2 toxin contaminated diets. Pathological signs of toxicity were investigated on days 21 and 39 of the trial, individual liveweight was measured weekly. Five birds from each group were sacrificed on the 21st and 39th days of treatment, when blood plasma, red blood cell, liver and kidney samples were taken, in which malondialdehyde and reduced glutathione concentration and glutathione-peroxidase activity were determined. Pathological signs (lesions in the oral cavity and on the tongue, inflammation in the small intestine) were found in the group fed T-2 and HT-2 toxin contaminated feed on day 21 compared to control. Body weight was significantly lower as a result of feeding T-2 and HT-2 toxin contaminated diet. However, the contamination did not cause a significant increase of malondialdehyde content in the analysed tissues. Reduced glutathione content was significantly lower in the liver homogenate on day 39 than that of the control. Glutathione peroxidase activity also did not differ significantly in blood plasma, red blood cell haemolysates and kidney homogenates, while it was significantly higher in the liver homogenates of the mycotoxin-challenged birds. In conclusion, it can be stated that T-2 and HT-2 toxin exposure has long-term effects in broiler chickens.


2015 ◽  
Vol 96 (5) ◽  
pp. 862-868
Author(s):  
A A Skal’nyy ◽  
A A Tin’kov ◽  
Yu S Medvedeva ◽  
I B Alchinova ◽  
E Yu Bonitenko ◽  
...  

Aim. To investigate the effect of zinc supplementation at physical exercise on the distribution of the metal in the tissues and the activity of serum antioxidant enzymes. Methods. Physical activity was simulated using the treadmill. Laboratory animals were distributed to 6 even (n=12) groups. The first and fourth groups of animals received no zinc-containing additives and were imposed to low and high physical activity, respectively. Animals of the 2 and 3 as well as 5 and 6 groups received 5 and 15 mg/kg/day of zinc asparaginate intragastrically and were imposed to low and high physical activity, respectively. The zinc concentrantion in the organs and tissues was determined by inductively coupled plasma mass spectrometry. The activity of antioxidant enzymes was determined by spectrophotometry. Results. Administration of zinc asparaginate to the laboratory animals with low physical activity resulted in a dose-dependent increase of the metal concentration in liver and kidney parenchyma and blood serum, as well as in increase of serum glutathione peroxidase activity. Intensive physical activity for 14 days was accompanied by a significant increase in serum and kidney tissue zinc level. At the 7-day exposure to zinc at physical activity, a dose-dependent increase in zinc concentration in the organs and increase of serum glutathione peroxidase activity was registered. Zinc administration together with physical activity for 14 days did not result in a significant change in the balance of metal in the body of animals. In contrast to the 7-day exposure, a combination of factors studied for 14 days was accompanied by increased activity of superoxide dismutase, but not glutathione peroxidase. Conclusion. Physical activity of different duration has a significant effect on the zinc kinetics at oral administration, and the activity of serum antioxidant enzymes in laboratory animals.


Author(s):  
K. Dvorshchenko ◽  
O. Korotkiy ◽  
D. Grebinyk ◽  
Ye. Torgalo

The goal of this work was the research of chondroitin sulfate prophylactic action on the glutathione system state in rat blood serum during acute joint inflammation. The research was performed on white non-linear pubertal rat males weighting 160-240 grams in compliance to all general ethical principles of animal research. All animals were divided to four experimental groups. The first group – the control: the animals were injected with 0,1 ml of 0,9 % NaCl solution in a right hind leg. The second group – rats were subjected to daily intramuscular injections of 3 mg/kg of chondroitin sulfate in the theurapeutic dose during 28 days. The third group – the animals were subjected to daily doses of 0,1 ml of 0,9 % NaCl solution injected in right hind extremities and starting from 29th day the acute joint inflammation was modelled (the animals were subjected to 0,1 ml of 1% of сarrageenan intramuscular injection in right hind extremities. The fourth group – rats were receiving intramuscular injections of therapeutic dose of 3 mg/kg of chondroitin sulfate for 28 days, and after that the acute joint inflammation was modelled starting from 29th day. 40 animals in all were taking part in the experimental research. The glutathione peroxidase activity was assessed judging from the decrease in GSH amount in the reaction with Ellman reagent. The glutathione peroxidase activity was estimated by the decrease in probe optical density owing to NADPH oxidation. Glutathione transferase activity was estimated by the speed of the conjugate formation between GSH and 1-chloro-2,4-dinitrobemzene. The amount of reduced glutathione was estimated spectrophotometrically using orthophthalic aldehyde. It was established that during the carrageenan-induced inflammation the glutathione peroxidase, glutathione reductase and glutathione reductase activity were reduced, a well as the amount of reduced glutathione, whereas the glutathione transferase activity was increased in comparison to the control. It was shown that under the prophylactic injection of the chondroitin sulfate based preparate to animals with acute joint inflammation the aforementioned parameters were partially stabilized.


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