scholarly journals Influence of L-Nω-nitroarginine methyl ester and sodium nitroprusside in vitro on the oxidative modification of rat lysosome proteins

2017 ◽  
Vol 98 (6) ◽  
pp. 1005-1011
Author(s):  
M A Fomina ◽  
A M Kudlaeva ◽  
S A Isakov ◽  
A N Ryabkov
Keyword(s):  
Methyl Ester ◽  
Sodium Nitroprusside ◽  
Adaptive Capacity ◽  
Oxidative Modification ◽  
Control Group ◽  
Total Level ◽  
Protein Oxidative Modification ◽  
In Vitro Incubation ◽  
Lysosomal Proteins

Aim. To investigate in vitro effects of 5 mM L-Nω-nitroarginine methyl ester and 0.1 mM sodium nitroprusside on oxidative modification of lysosomal proteins of liver of intact sexually mature female rats of Wistar line. Methods. In the control groups in vitro incubation of isolated lysosomes in the isolation medium for 1, 2 and 4 hours was carried out. Experimental groups were incubated similarly in solutions of 5 mM L-Nω-nitroarginine methyl ester and 0.1 mM sodium nitroprusside. Protein oxidative modification was measured in sedimentary fraction according to R.L. Levine’s method in E.E. Dubinina’s modification. Reserve-adaptive capacity was calculated as the difference between total area under the curve of carbonyl derived proteins after metal-catalyzed oxidation (taken as 100%) and spontaneous oxidation, expressed as a percentage. Results. After 4-hour in vitro incubation 5 mM L-Nω-nitroarginine methyl ester was found to statically significantly increase the total level of protein oxidative modification compared to the control group by 2.41 times and to reduce reserve-adaptive capacity by 4.96 times, and 0.1 mM sodium nitroprusside increases the total level of protein oxidative modification compared to the control group by 2.05 times and reduces reserve-adaptive capacity by 1.56 times. One of the possible mechanisms of this phenomenon may be the reduced activity of lysosomal proteinases. 2-hour and 4-hour in vitro incubation of lysosomes in 5 mM L-Nω-nitroarginine methyl ester is accompanied by an increase of secondary markers of the ratio of protein oxidative modification relatively to 1-hour exposure by 1.18 times and 1.35 times, respectively. At 1-hour in vitro incubation in 0.1 mM sodium nitroprusside, increase of secondary markers of protein oxidative degradation by 1.64 times occurs. Conclusion. The in vitro effect of 5 mM -Nω-nitroarginine methyl ester and 0.1 mM sodium nitroprusside results in visible changes of oxidative modification of rat liver lysosomal proteins.

scholarly journals Protein oxidative modification and cathepsin H activity in rats’ thymocytes at nitrogen oxide (II) synthesis modulation in vitro

2014 ◽  
Vol 95 (4) ◽  
pp. 553-557 ◽  
Author(s):  
Yu V Abalenikhina ◽  
M A Fomina
Keyword(s):  
Methyl Ester ◽  
Nitrogen Oxide ◽  
Nutrient Medium ◽  
No Synthase ◽  
Oxidative Modification ◽  
Control Group ◽  
Griess Reagent ◽  
Protein Oxidative Modification ◽  
Cathepsin H

Aim. To study the influence of substrate for nitrogen oxide (II) synthesis - L-arginine - and non-selective NO-synthase inhibitor - N-nitro-L-arginine-methyl ester - on protein oxidative modification in combination with rats’ thymocytes cathepsin H activity estimation in vitro. Methods. The study was performed on male Wistar rats with body weight of 280-320 g. Freshly-separated thymocytes were incubated in vitro in the full nutrient medium containing 5 мМ of N-nitro-L-arginine-methyl ester (n=8) or 5 мМ of L-arginine for 24 hours at the temperature of 37 °C (n=8). Control group consisted of thymocytes incubated in the same conditions in the full nutrient medium (n=8) for 24 hours. Nitric oxide metabolites levels were measured by spectrophotometry in the visible spectrum using the reaction with Griess reagent. Cathepsin H activity was estimated by BarrettKirschke spectrofluorimetry. Protein oxidative modification was measured by R.L. Levine method in E.E. Dubinina modification followed by carbonyl derivatives absorption spectrum quantitative analysis. Results. In nitrogen oxide (II) synthesis deficiency model, protein oxidative modification degree increased, mainly due to basic and neutral aldehyde- and ketone-dinitrophenylhydrazones level increase. Those changes were accompanied by increased activity of cathepsin H. In nitrogen oxide (II) synthesis stimulation model, level of oxidative-modified proteins decreased, mainly due to lower levels of neutral amino acid derivatives, cathepsin H activity didn’t change. Conclusion. In vitro nonselective inhibitor of inducible NO-synthase - N-nitro-L-arginine-methyl ester - stimulates protein oxidative modification and increases activity of cathepsin Н; substrate of NO synthesis - L-arginine - showes antioxidant effect.

322 EFFECT OF SODIUM NITROPRUSSIDE ON BUFFALO SPERM CAPACITATION IN VITRO

2007 ◽  
Vol 19 (1) ◽  
pp. 276 ◽  
Author(s):  
L. Boccia ◽  
L. Attanasio ◽  
A. De Rosa ◽  
G. Pellerano ◽  
R. Di Palo ◽  
...  
Keyword(s):  
Sodium Nitroprusside ◽  
Acrosome Reaction ◽  
Production Efficiency ◽  
Control Group ◽  
Sperm Capacitation ◽  
Promoting Effect ◽  
Vitro Fertilization ◽  
Domestic Species ◽  
Frozen Semen

The overall in vitro embryo production efficiency in buffalo is hampered by the poor fertilization rate. It is known that the quality of the frozen semen may affect fertilization efficiency. However, it is not possible to rule out that the process of capacitation, required by spermatozoa to acquire the fertilizing ability, is impaired in the in vitro fertilization (IVF) system. Although several agents have been proven to induce sperm capacitation in vitro, heparin treatment is still the most efficient method in most of the domestic species. There is evidence that capacitation is part of an oxidative process and that nitric oxide (NO) acts as a capacitation inducer in human (Herrero et al. 1999 Biol. Reprod. 61, 575–581) and bovine (Rodriguez et al. 2005 Anim. Reprod. Sci. 85, 231–242) spermatozoa. The aim of the present study was to evaluate whether sodium nitroprusside (SNP), a well-known generator of NO in vitro, improves buffalo sperm capacitation in vitro. Frozen–thawed sperm from a bull previously tested for IVF were treated by swim-up in order to select only the motile population. Spermatozoa were incubated in the presence of 0.01 mM heparin (control group) for 1 h (n = 266), 2 h (n = 270), and 3 h (n = 306), and in the presence of 10 �M SNP for 1 h (n = 302), 2 h (n = 286), and 3 h (n = 260). The concentration of SNP was chosen on the basis of a preliminary dose-response trial (0.1 �M, 1 �M, and 10 �M). Following incubation with these agents, sperm were exposed for 15 min to 60 �g mL-1 of lysophosphatidylcholine, an agent known to induce acrosome reaction only on capacitated spermatozoa. Trypan blue was used first to differentiate live from dead spermatozoa and the dried smears were then fixed in 37% formaldehyde and stained with Giemsa for acrosome evaluation by microscopic examination. The proportion of acrosome-reacted spermatozoa in each group was used to assess the efficiency of capacitation under different incubation conditions. Differences between groups were analyzed by chi-squared test. No dead spermatozoa were found in all groups. Following 1-h sperm treatment with either heparin or SNP, the proportion of acrosome-reacted spermatozoa was similar (35.3% vs. 28.5%, respectively). However, extending the incubation time to 2 h, SNP significantly (P < 0.01) increased the incidence of acrosome reaction compared to heparin (60.1% vs. 44.1%, respectively). Analogously, when the sperm treatment was prolonged to 3 h, SNP gave a significantly (P < 0.01) higher percentage of acrosome reaction compared to the control (68.8% vs. 36.6%, respectively). In conclusion, sperm treatment with SNP for either 2 or 3 h significant improved the efficiency of buffalo sperm capacitation in vitro compared with heparin, that is, the capacitating agent currently used in the IVF system. The promoting effect of SNP indirectly indicates that NO acts as a capacitation inducer in buffalo spermatozoa. Finally, these results suggest the need to evaluate the effect of SNP on the fertilizing capability of buffalo spermatozoa in vitro.

scholarly journals PPARα/γ, adiponectin and GLUT4 overexpression induced by moronic acid methyl ester influenced on glucose and triglycerides levels of experimental diabetic mice

2021 ◽  
Author(s):  
Samuel Estrada-Soto ◽  
Litzia Cerón-Romero ◽  
Gabriel Navarrete-Vázquez ◽  
Edgar Rosales-Ortega ◽  
Jaime H. Gómez-Zamudio ◽  
...  
Keyword(s):  
Methyl Ester ◽  
Acid Methyl Ester ◽  
Control Group ◽  
Oral Glucose ◽  
Acute Administration ◽  
Diabetic Mice ◽  
Ppar Γ ◽  
Acid Methyl

The current study aimed to determine the antidiabetic and antidyslipidemic activities of moronic acid methyl ester (1) by in vivo, in vitro, in silico and molecular biology studies. Compound 1 was evaluated to establish its dose-dependent antidiabetic and antihyperglycemic (50 mg/kg) activities, in diabetic and normoglycemic male CD1 mice, respectively. Also, compound 1 was subjected to a sub-acute study (50 mg/kg/day for eight days) to determine blood biochemical profiles and the expression of PTP-1B, GLUT4, PPAR-α, PPAR-γ, adiponectin, IL-1β, and MCP1 in adipose tissue of animals after treatment. Different doses in acute administration of 1 decreased glycemia (p < 0.05), compared with vehicle, showing greater effectiveness in the range 50-160 mg/kg. Also, the oral glucose tolerance test (OGTT) showed that 1 induced a significant antihyperglycemic action by opposing the hyperglycemic peak (p < 0.05). Moreover, 1 subacute administration decrease glucose and triglycerides levels after treatment (p < 0.05); while the expression of PPAR-α and γ, adiponectin and GLUT4 displayed an increase (p< 0.05) compared with the diabetic control group. In conclusion, compound 1 showed antihyperglycemic, antidiabetic and antidyslipidemic effects in normal and diabetic mice, probably due to insulin sensitization through increase mRNA expression of GLUT4, PPAR-α, PPAR-γ and adiponectin genes.

scholarly journals THE INFLUENCE OF THYROID HORMONES ON PROTEIN OXIDATIVE MODIFICATION IN CASE OF EXPERIMENTAL PERIODONTITIS

2019 ◽  
pp. 52-59
Author(s):  
V. V. Shcherba ◽  
T. Ya. Yaroshenko ◽  
R. M. Kubant ◽  
M. M. Korda
Keyword(s):  
Thyroid Hormones ◽  
Blood Serum ◽  
Periodontal Diseases ◽  
Oxidative Modification ◽  
Control Group ◽  
Periodontal Tissues ◽  
Protein Oxidative Modification ◽  
White Rats ◽  
Experimental Periodontitis ◽  
Derivatives Of

Introduction. The inflammatory-dystrophic process in periodontium is accompanied by its hypoxia, which causes the activation of free radical oxidation processes. Tissue destruction in periodontal diseases is considered to be the result of an altered inflammatory/immune response to microbial plaque and involves massive release of neutrophils, reactive oxygen species and enzymes. The aim of the study – to establish the protein oxidative modification in blood and periodontium in case of periodontitis combined with hyper- and hypothyroidism in rats. Research Methods. Experimental studies were carry out on 48 mature male, nonliner, white rats, which were divided into the following groups: І – control animals; ІІ – animals with periodontitis; ІІІ – animals with periodontitis combined with hyperthyroidism; IV – animals with periodontitis combined with hypothyroidism. Protein oxidative modification (POM) was determined in blood serum and periodontium homogenate by the method of I.F. Meschyshyn. Results and Discussion. The results of our studies showed that the intensity of the processes of protein oxidative modification was significantly increased in animals of all experimental groups vs control group. The content of aldehyde- and ketone derivatives of the alkali nature changed more pronounced than the content of aldehyde and ketone derivatives of neutral nature. Conclusions. Thus, the experimental periodontitis is accompanied by a marked increase in the intensity of the protein oxidative modifications both in the homogenate of periodontal tissues and in the blood serum. Imbalance of thyroid hormones increases oxidative stress in experimental periodontitis, especially pronounced in hyperthyroidism.

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