The rat blood plasma was obtained by centrifugation of whole blood and immediately used to determine biochemical markers of the functional state of the liver (the activity of alanine aminotransferase and aspartate aminotransferase, bilirubin concentration, and the thymol test) and lipid concentrations2 (link). ALT and AST activities were determined by Reitman-Fresnel method, total and direct bilirubin were determined by Endraschik method and thymol test was performed using thymol reagent checking the test kits (R&D enterprise Felicity-Diagnostics, Ukraine).
We determined the concentration of the following compounds in the blood plasma (mg%): phospholipids, cholesterol (CHOL), cholesterol esters (ECHOL), free fatty acids, triglycerides. Lipids were divided by the method of thin-layer chromatography49 . Chromatographic separation of lipid components of plasma was carried out on “Silufol” plates. After treatment with an aqueous solution of phosphomolybdic acid, a quantitative assessment of the color intensity of each fraction was performed using a densitometer DO-1 M (“Shimadzu”, Japan, λ 620 nm)50 (link).
Blood cell mass was used to obtain erythrocyte plasma membrane preparations by the slightly modified Dodge’s method. Plasma membrane preparations were used to determine the ATPase activities of the primary active ion transport systems (total Mg2+, Na+, K+-ATPase, basal Mg2+-ATPase and Na+, K+-ATPase). The protein concentration in the preparations of the erythrocyte plasma membranes (PM) was determined by Lowry’s method51 (link). Total Mg2+, Na+, K+-ATPase activity was determined in the fraction of erythrocyte PMs in the standard incubation medium (in mM): 1 ATP, 3 MgCl2, 125 NaCl, 25 KCl, 1 EGTA, 20 Hepes-Tris-buffer (pH 7.4), 1 NaN3 (inhibitor of mitochondria ATPase), 0.1 µm thapsigargin (the selective inhibitor of Ca2+,Mg2+-ATPase of endoplasmatic reticulum) and 0.1% digitonin (the factor of PM perforation), at 37 °C. The Mg2+-ATPase activity was determined by the presence of a selective inhibitor Na+,K+-ATPase ouabain (1 mM) in the incubation medium. The Na+, K+-ATP activity was calculated as the difference between the total Mg2+, Na+, K+-ATPase and the ouabain-insensitive Mg2+-ATPase activity52 (link),53 (link).
This paper presents a statistical analysis of the experimental data obtained in the study and processed by the variation statistics methods using the Origin Pro 8 software. The samples were checked to belong to normally distributed general populations according to the Shapiro–Wilk criterion. The dispersion analysis was used to determine reliable differences between the mean values of samplings, and the post-test comparison was made using the Tukey test. In all cases, the results were reliable on the condition of the probability value p under 5% (p < 0.05). The obtained results were presented as the arithmetic mean ± standard error of the mean value, and the n value was determined by the total in the number of experiments.
We determined the concentration of the following compounds in the blood plasma (mg%): phospholipids, cholesterol (CHOL), cholesterol esters (ECHOL), free fatty acids, triglycerides. Lipids were divided by the method of thin-layer chromatography49 . Chromatographic separation of lipid components of plasma was carried out on “Silufol” plates. After treatment with an aqueous solution of phosphomolybdic acid, a quantitative assessment of the color intensity of each fraction was performed using a densitometer DO-1 M (“Shimadzu”, Japan, λ 620 nm)50 (link).
Blood cell mass was used to obtain erythrocyte plasma membrane preparations by the slightly modified Dodge’s method. Plasma membrane preparations were used to determine the ATPase activities of the primary active ion transport systems (total Mg2+, Na+, K+-ATPase, basal Mg2+-ATPase and Na+, K+-ATPase). The protein concentration in the preparations of the erythrocyte plasma membranes (PM) was determined by Lowry’s method51 (link). Total Mg2+, Na+, K+-ATPase activity was determined in the fraction of erythrocyte PMs in the standard incubation medium (in mM): 1 ATP, 3 MgCl2, 125 NaCl, 25 KCl, 1 EGTA, 20 Hepes-Tris-buffer (pH 7.4), 1 NaN3 (inhibitor of mitochondria ATPase), 0.1 µm thapsigargin (the selective inhibitor of Ca2+,Mg2+-ATPase of endoplasmatic reticulum) and 0.1% digitonin (the factor of PM perforation), at 37 °C. The Mg2+-ATPase activity was determined by the presence of a selective inhibitor Na+,K+-ATPase ouabain (1 mM) in the incubation medium. The Na+, K+-ATP activity was calculated as the difference between the total Mg2+, Na+, K+-ATPase and the ouabain-insensitive Mg2+-ATPase activity52 (link),53 (link).
This paper presents a statistical analysis of the experimental data obtained in the study and processed by the variation statistics methods using the Origin Pro 8 software. The samples were checked to belong to normally distributed general populations according to the Shapiro–Wilk criterion. The dispersion analysis was used to determine reliable differences between the mean values of samplings, and the post-test comparison was made using the Tukey test. In all cases, the results were reliable on the condition of the probability value p under 5% (p < 0.05). The obtained results were presented as the arithmetic mean ± standard error of the mean value, and the n value was determined by the total in the number of experiments.
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