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Objective To investigate the effects of ω-3 polyunsaturated fatty acid(PUFA)on oxidative stress produc-tion and apoptosis in hippocampus of neonatal rats with brain injury induced by lipopolysaccharide(LPS). Methods Forty-eight neonatal Sprague Dawley rats(3 days)were randomly divided into control group,LPS group,ω-3 PUFA group and ω-6 PUFA group,with 12 rats in each group. The rats in the LPS group,ω-3 PUFA group and ω-6 PUFA group were given 0. 6 mg·kg - 1 LPS via intraperitoneal injection,then equal volume of saline,ω-3 PUFA and ω-6 PUFA was immediately given via intraperitoneal injection respectively;while the rats in the control group were all given equal volume of saline. The rats in each group were sacrificed at 24 hours after intraperitoneal injection with saline or fat emulsions to obtain the hippocampus. The levels of superoxide dismutase(SOD)and malondialdehyde(MDA),reduced glutathion(GSH),oxidized glutathione(GSSG) were detected and GSSG/ GSH was calculated. The apoptotic index was measured by terminal deoxynucleotidyl transferase-me-diated dUTP nick end labeling. Results Compared with the control group,the levels of SOD and GSH in hippocampus of rats in the LPS group,ω-6 PUFA group and ω-3 PUFA group were significantly decreased(P < 0. 05),and the levels of MDA, GSSG and the ratio of GSSG/ GSH were significantly increased(P < 0. 05). Compared with the LPS group,the levels of SOD and GSH in hippocampus of rats in ω-6 PUFA group were significantly decreased(P < 0. 05),the levels of MDA,GSSG and the ratio of GSSG/ GSH were significantly increased(P < 0. 05);the levels of SOD and GSH in hippocampus of rats in ω-3 PU-FA group were significantly increased(P < 0. 05),the levels of MDA,GSSG and the ratio of GSSG/ GSH were significantly de-creased(P < 0. 05). Compared with the ω-6 PUFA group,the levels of SOD and GSH in hippocampus of rats in ω-3 PUFA group were significantly decreased(P < 0. 05),the levels of MDA,GSSG and the ratio of GSSG/ GSH were significantly in-creased(P < 0. 05). The apoptotic index in the LPS group,ω-6 PUFA group and ω-3 PUFA group was higher than that in the control group(P < 0. 05). The apoptotic index in the ω-6 PUFA group was higher than that in the LPS group(P < 0. 05). The apoptotic index in the ω-3 PUFA group was lower than that in the LPS group and ω-6 PUFA group(P < 0. 05). Conclusion ω-3 PUFA can alleviate the oxidative stress,and decrease the apoptosis of hippocampus in neonatal rats with brain injury in-duced by LPS. So it has a neuroprotective effect in brain injury induced by LPS.
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Objective To compare the level of glutathione (GSH) and oxidized glutathione (GSSG), the ratio of GSH/GSSG and the concentration of albumin in plasma of patients with complicated and un-complicated falciparum malaria. Methods This research was a cross sectional study using comparison analysis with the plasma GSH and GSSG, the ratio of plasma GSH/GSSG and the concentration of plasma albumin as variables. The complicated malaria patients were obtained from Dr. Saiful Anwar Hospital Malang, whereas uncomplicated malaria patients were obtained from the Regency of Pleihari South Kalimantan. Plasma GSH and GSSG levels were determined by the spectrophotometer at the wave length of 412 nm, whereas the concentration of albumin was determined by bromocresol green method in the pH of 4.1. Results There were no significant differences between the level of plasma GSH and GSSG in complicated and uncomplicated malaria patients, as well as the ratio of plasma GSH/GSSG in the two groups (P = 0.373; P = 0.538; and P = 0.615, respectively, independent t-test). In contrast, the plasma albumin concentration in complicated malaria patients were significantly higher than uncomplicated malaria patients (P = 0.000, Mann Whitney U test). Conclusions It can be concluded that the average of plasma GSH and GSSG level, also plasma GSH/GSSG ratio in complicated malaria are not different from uncomplicated malaria. Although plasma concentration of albumin in both groups is below the normal range, there is an increase in complicated malaria that might be as compensation of oxidative stress.
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Objective: To compare the level of glutathione (GSH) and oxidized glutathione (GSSG), the ratio of GSH/GSSG and the concentration of albumin in plasma of patients with complicated and un-complicated falciparum malaria. Methods: This research was a cross sectional study using comparison analysis with the plasma GSH and GSSG, the ratio of plasma GSH/GSSG and the concentration of plasma albumin as variables. The complicated malaria patients were obtained from Dr. Saiful Anwar Hospital Malang, whereas uncomplicated malaria patients were obtained from the Regency of Pleihari South Kalimantan. Plasma GSH and GSSG levels were determined by the spectrophotometer at the wave length of 412 nm, whereas the concentration of albumin was determined by bromocresol green method in the pH of 4.1. Results: There were no significant differences between the level of plasma GSH and GSSG in complicated and uncomplicated malaria patients, as well as the ratio of plasma GSH/GSSG in the two groups (P=0.373;P=0.538;and P=0.615, respectively, independent t-test). In contrast, the plasma albumin concentration in complicated malaria patients were significantly higher than uncomplicated malaria patients (P=0.000, Mann Whitney U test). Conclusions: It can be concluded that the average of plasma GSH and GSSG level, also plasma GSH/GSSG ratio in complicated malaria are not different from uncomplicated ma-laria. Although plasma concentration of albumin in both groups is below the normal range, there is an increase in complicated malaria that might be as compensation of oxidative stress.
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Objective:To establish an HPLC method for the determination of oxidized glutathione in glutathione tablets. Meth-ods:AnAgilentTC-C18(250mm×4.6mm,5 μm)columnwasusedwiththemobilephaseofphosphatesolution(pH3.0)-methanol (96∶4). The flow rate was 1. 0 ml·min-1. The detection wavelength was 210 nm. The column temperature was 30℃ and the injec-tion volume was 20 μl. Results:The linear range of oxidized glutathione was 1.403-22.450 μg·ml-1(r=0.999 9). The recovery was 98. 9% with RSD of 1. 5%(n=9). Conclusion:The method is accurate and repeatable, and can control the content of oxidized glutathione in glutathione tablets effectively. .
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El glutatión (GSH) es una molécula única que participa en aspectos esenciales de la homeostasis celular, teniendo un rol central en la defensa contra el daño oxidativo. El GSH (L-g-glutamil-L-cisteinil-glicina) es un tripéptido hidrosoluble formado por los aminoácidos ácido glutámico, cisteína y glicina que se encuentra presente en el citoplasma de todas las células. La forma oxidada de la molécula, GSSG, se encuentra principalmente en forma extracelular. Las concentraciones de GSH y GSSG y su relación molar son indicadores de la funcionalidad celular y su alteración está relacionada con varios procesos patológicos en el hombre y en los animales de compañía. En esta revisión se abordan importantes aspectos de la homeostasis, las principales funciones biológicas y las metodologías analíticas disponibles para el análisis de GSH en sangre y plasma.
Glutathione is a unique molecule that participates in key cellular homeostasis, having a central role in defense against oxidative damage. GSH (L-g-glutamyl-L-cysteinyl-glycine) is a water soluble tripeptide composed of amino acid glutamic acid, cysteine and glycine. GSH is present in every cell cytoplasm. The oxidized form of the molecule, GSSG, is found primarily in extracellular form. GSH and GSSG concentrations and their molar ratio are indicators of cell function and its alteration is associated with several disease processes in humans and in companion animals. This review focuses on important aspects of homeostasis, major biological functions and available analytical methodologies for the analysis of GSH in blood and plasma.
A glutationa (GSH) é urna molécula única envolvida em aspectos essenciais da homeostase celular, tendo um papel central na defesa contra o dano oxidativo. O GSH (L-g-glutamil-L-cisteinil-glicina) é um tripeptídeo hidrossolúvel formado pelos aminoácidos: ácido glutámico, cisteína e glicina que se encontra presente no citoplasma de todas as células. A forma oxidada da molécula, GSSG, acha-se principalmente em forma extracelular. As concentragoes de GSH e GSSG e a sua relagáo molar sao indicadores da funcionalidade celular e a sua alteragao está relacionada com vários processos patológicos no homem e nos animais de estimagáo. A presente revisáo aborda questoes importantes da homeostase, as principais fungoes biológicas e as metodologias analíticas disponíveis para a análise de GSH em sangue e plasma.
Subject(s)
Humans , Glutathione/blood , Homeostasis , Homeostasis/physiology , Hemostatic Techniques , Oxidative Stress , PlasmaABSTRACT
Glutathione-S-transferase (EC 2.5.1.18) activity was assayed in hepatic and extrahepatic tissues of pigeons using l-chloro-2,4-dinitrobenzene and l,2-dichloro-4-nitrobenzene as substrates. Gluthathione-S-transferase activity towards l-chloro-2,4-dinitrobenzene in pigeon was in the order: kidney >liver >testes >brain >lung>heart. The enzyme activity with 1- chloro-2,4-dinitrobenzene as substrate was 40-44 times higher in pigeon liver and kidney than that observed with l,2-dichloro-4-dinitrobenzene as substrate. Km values of hepatic and renal glutathione transferase with l-chloro-2,4-dinitrobenzene as substrate were 2.5 and 3 mM respectively. Double reciprocal plots with varying reduced gluthathione concentrations resulted in biphasic curves with two Km values (liver 0.31 mM and 4mM; kidney 0.36 mM and 1.3 mM). The enzyme activity was inhibited by oxidized gluthathione in a dose-dependent pattern. 3-Methylcholanthrene elicited about 50% induction of hepatic glutathione transferase activity whereas phenobarbital was ineffective.