Tag Archives: ethanol

Different sensitivity of Na(+),K(+)-ATPase and Mg(2+)-АТРase to ethanol and arachidonic acid in rat colon smooth muscle under pretreatment of cellular membranes with Ds-Na

A. A. Kaplia

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: kaplya@biochem.kiev.ua

The methodological procedure provides the detection of the relatively high Na+,K+-ATPase functional activity in the crude cellular membranes of rat colon smooth muscle (CSM) following standard detergent pretreatment (with Ds-Na vs digitonin). It includes the essential discrete steps: detergent membrane permeabilization under optimal detergent/protein ratio and active site protection by ATP (for Ds-Na) prior enzymatic reaction with substantial detergent dilution far below critical micelle concentration in the ATPase medium. The high level of the Na+,K+-ATPase activity, originally detected in CSM, did not differ for two detergents and was comparable with ouabain-resistant Mg2+,АТР-hydrolase activity, The features of ATPase protein-lipid complexes were evaluated by the enzyme sensitivity to the effect of ethanol and arachidonic acid with different membrane disordering effectiveness. The long-chain fatty acid is a more effective inhibitor as compared with aliphatic alcohol for both ATPases. Mg2+,АТР-hydrolase appeared to be much more resistant to inactivation than Na+,K+-ATPase. The data reflect the possible differences in lipid dependence of two enzymatic systems due to the peculiarities of the structural arrangement in membrane and importance of the hydrophobic microenvironment for mechanism of catalysis. Thus, the data represent the approach to the simple and reliable Na+,K+-АТРase activity determination in nonpurified CSM membranes, acceptable for different tissues and appropriate for quantitative comparison in pathophysiological studies and for testing the impact of diverse  effectors on Na+,K+-АТРase.

Alterations of prooxidant-antioxidant system of rat liver at ethanol and tetracycline action

Kh. Yu. Nedoshytko

I. Ya. Horbachevsky Ternopil State Medical University, Ukraine;
e-mail: khrystynan@ukr.net

The state of antioxidant system and fatty acid composition of lipids in the liver tissues of rats of different sex at the ethanol and tetracycline action and at the influence of biologically active additives (BAA) “Alpha + Omega” at a dose of 0.5 mg/kg b.w. per os was investigated. It was found that the contet of lipid peroxidation products in the liver was increased at the action of 40 % ethanol at a dose of 7 ml/kg b.w. per os and tetracycline – 500 mg/kg and more profound at their joint using. Howe­ver, the content of diene conjugates was stronger increased in the liver of females at the action of ethanol, while in the liver of males at the action of tetracycline (Р < 0.05). It was shown that the application of the investigated compounds led to the reduction of an antioxidant defense system activity of males and females liver, as evidenced by the decrease of superoxide dismutase activity by 46 and 43% and reduction of glutathione content by 39 and 38% (Р < 0.05). The activity of alanineaminotransferase, aspartateaminotransferase and alkalinephosphatase was increased in the liver of males and females under the influence of ethanol and tetracycline and more profound at their joint usa­ge (Р < 0.05). It was established that ethanol and tetracycline unidirectionally changed fatty acid composition of total lipids of rat liver, but at the ethanol action the changes were more expressed in females while at the tetracycline action in males. The application during 14 days of BAA “Alpha + Omega” to male and female rats with an acute tetracycline damage at subacute ethanol action led to partial normalization of prooxidant-antioxidant system and the relative content of total lipids fatty acids of the liver of both sexes animals.

Actomyosin ATPase activity of skeletal muscles and the markers of tissue damage in the blood of rats under prolonged chronic alcoholization

Yu. V. Tseyslyer1, О. M. Podpalova2, N. Е. Nurishchenko1, V. S. Маrtyniuk1

1ESC Institute of Biology, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: yuliya.tseysler@gmail.com,
2Bogomolets National Medical University, Kyiv, Ukraine;
e-mail: olgapodpalova@gmail.com

The activity of creatine kinase and indices of lipid metabolism in the blood and also actomyosin ATPase activity of skeletal muscles of rats under chronic 8-month alcohol abuse were investigated. It is shown that actomyosin K+-ATPase activity of skele­tal muscles increases from two months of ethanol use, but actomyosin Mg2+-ATPase activity decreases during 6-8 months of alcoholization. From two months of ethanol use the creatine kinase activity, as an enzyme marker of muscle tissue damage, statistically significantly increases during all the period of the animals alcoholization. The level of total lipid increases after two months of alcohol consumption (in blood plasma by 30% and in erythrocyte mass by 65%). For longer periods of alcoholization (4-8 months) the level of lipids remains almost the same, whereas in erythrocyte mass it does not differ from control values. The level of diene conjugates in the blood plasma reduces and the amount of ketone derivatives of fatty acid residues increases that points to the inhibition of some components of the antioxidant system that control detoxification of hydroperoxides of fatty acids and also to activation of free radical damage of tissues. There were no significant changes of lipid peroxidation level in erythrocyte mass at any stage of alcoholization.