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ENGLISH VERSION: BIOENERGETIC METABOLISM STATE IN ALBINO RATS UNDER THE LONG-LASTING EXPOSURE OF THE LAPROXIDES SUBTOXIC DOSES*
Kucheryavchenko M.A, Zaitseva O. V., Zhukov V.I., Knigavko V.G. Kharkiv National Medical University, Kharkiv, Ukraine
It was investigated the long-lasting exposure (1,5 months) of L-303 and L-500 laproxides in doses of 1/10, 1/100, 1/1000 LD50 on metabolism of the macroergic compounds and their metabolites in albino rats liver by such indexes: content of adenosintriphosphate (ATPh), adenosindiphosphate (ADPh), adenosinmonohosphate (AMPh), cyclic guano-sinmonophosphate (cGMPh), inorganic phosphate, creatinphosphate, adenine nucleotides as well as Ca2+- and Mg2+-ATPh-ase activity. It was determined the inhibition of bioenergeticprocesses, catabolism predominance over restoration syntesis in albino rats organism under L-303 and L-500 laproxides long-lasting exposure. These xenobiotics in 1/10 and 1/100 LD50 doses decreased the content of ATPh, ADPh, cGMPh, adenine nucleatides, creatinphosphate, value of cell energetic potential, reduced Mg2+- ATPh-ase, Ca2+-ATPh-ase activtty. Negative effect of L-303 and L-500 laproxides on macroergic compounds metabolism in liver manifestated in augmentation AMPh, cAMPh, inorganic phosphate levels with respect to control. It wasn't detect statistic differences between obtained results in experimental groups and control under laproxides effect in 1/1000 LD50 dose. Desorders in balance of bioenergetic metabolism indexes in albino rats organism confirm the presence of laproxides hepatotoxic effect which causes subsequent development metabolic desorders.
Key words: laproxides, macroergic compounds, albino rats, subacute toxicologic experiment.
Introduction
Development of the chemical industry is accompanied by an increase in the production of chemicals to which people are not evolutionarily adapted. Accumulation of xenobiotics in the environment and their devastating effect on flora and fauna often generates environmentally caused diseases and pathological conditions [1,3]. Numerous chemical compounds have not only a direct toxic effect, but also the ability to influence the development of long-term effects: carcinogenesis, mutagenesis, teratogenic effect, atherogenesis, immunological deficiency, accelerated aging, etc. [2,6]. Today, one of the most powerful sources of pollution of the biosphere are enterprises of synthetic organic chemistry. This fully applies to the chemical plant for the production of "Laproxides". These chemicals are widely used to produce epoxy resins, lacquers, enamels, paints, etc. and have found application in many sectors of the economy - construction, engineering, electrochemistry, oil production and agriculture.
However, the absence of comprehensive characteristics of the potential danger of these compounds for human health and the environment dictates the need for a thorough study of the mechanisms of laproxides biological action and develop of metabolic disturbances correction ways that occur under the influence of the xenobiotics small data sub-toxic doses.
The aim of the work was to study the long-term impact of sub-toxic doses of the laproxides new group on the bioenergetic metabolism state in toxicological experiment.
Materials and methods
In this study we used a new group of laproxides with regulated physicochemical properties. These compounds are related to the class of polyethers: oligoethermonoepoxid with molecular weight of 500 (L-500) and triglycidyl ether polioksipropilentriol with molecular weight of 303 (L-303). With respect to the results of acute experiments, these substances have a low toxicity and weak cumulation, have not the species
and gender sensitivity. The mean lethal doses (LD50) of L-303 and L-500 for white rats are set at levels of 5.75 g/kg and 26.7 g/kg of body weight of the animal, and the coefficients of cumulation (Kk) were 7.61 and 9.28. The research program included a long subacute toxicological experiment on mature white Wistar rats weighting 0.19-0.20 kg. Under the experimental conditions, the animals for 1.5 months every morning before feeding with a metal probe orally were administered aqueous solutions of laproxides doses of 1/10; 1/100; 1/1000 of LD50 (6 groups of n = 10 animals). The control group (n = 10 animals) received the appropriate volume of drinking water. In the experiment we strictly met the requirements of the bioethics and the principles of the "European Convention for the Protection of Vertebrate Animals used for experimental and other scientific purposes" (Strasbourg, 1986) [9]. Upon completion of the subacute experiment investigated the macroergic compounds metabolism and their metabolites in the liver, were moreover, it the content of the adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), inorganic phosphate, phosphocreatine, the amount of adenine nucleotides and the activity of Ca2+- and Mg2+ -dependent ATP-ase were determined. Determination of Ca2+- and Mg2+- dependent ATP-ase in rat hepatocytes was performed by conventional biochemical method [4]. ATP content in liver tissues was determined by the method of E. Beutler [8], ADP - by D. Jaworek [10], phosphocreatine - by ED Sonnin [5], an inorganic phosphate - on the method described by N. Meshkova and S. Severin [4]. The magnitude of the energy potential (EP) was calculated as D.E. Atrinson [7]. CAMP and cGMP content in the liver was determined by Ch. W. Parker [11]. The results obtained were processed by methods of variation statistics using Student -Fisher t-test.
Results and discussion
Study of the L-303 and L-500 laproxides effect at subtoxic doses of 1/10 and 1/100 of LD50 in terms long subacute experiment revealed liver of reduction of ATP, ADP, cGMP content, the amount of adenine nucleotides,
* To cite this English version: Kucheryavchenko M.A, Zaitseva O. V., Zhukov V.I., Knigavko V.G. Bioenergetic metabolism state in albino rats under the long-lasting exposure of the laproxides subtoxic doses / / Problemy ekologii ta medytsyny. - 2014. - Vol 18, № 1-2. - P. 47-49.
Tom. 18, N 1-2 2014 p.
phosphocreatine, the energy potential of the cell and the phosphate and cAMP (Table), when compared with the activity of Ca2 +- and Mg2 +- dependent ATP-ase agains a results of the control group. background increased AMP levels, the inorganic
Table
Influence of L-303 and L-500 laproxides at sub-toxic doses on indicators of bioenergy metabolism in white rats in subacute experiment
Indicators Monitoring Group, the dose of LD50, M ± m
Brand of laproxides Control (n = 10) 1/10 (n = 10) 1/100 (n = 10) 1/1000 (n = 10)
ATP (mcmol/g liver) L-303 2,24 ± 0,12 0,53 ± 0,04* 0,71 ± 0,09* 2,27 ± 0,16
L-500 2,24 ± 0,12 0,48 ± 0,04* 0,69 ± 0,03* 2,23 ± 0,14
ADP (mcmol/g liver) L-303 1,28 ± 0,07 0,46 ± 0,04* 0,53 ± 0,04* 1,26 ± 0,05
L-500 1,28 ± 0,07 0,41 ± 0,03* 0,5 ± 0,04* 1,26 ± 0,06
AMP (mcmol/g liver) L-303 0,82 ± 0,06 1,73 ± 0,14* 1,56 ± 0,08* 0,83 ± 0,07
L-500 0,82 ± 0,06 1,65 ± 0,13* 1,58 ± 0,12* 0,85 ± 0,05
Inorganic phosphorus (mcmol/g liver) L-303 5,79 ± 0,64 13,8 ± 1,2* 9,38 ± 0,74* 5,68 ± 0,54
L-500 5,79 ± 0,64 14,8 ± 1,27* 9,52 ± 0,87* 5,66 ± 0,47
cAMP (nmol/g liver) L-303 650,4 ± 27,2 935,4 ± 41,6* 896,2 ± 38,5* 640,2 ± 31,4
L-500 650,4 ± 27,2 940,6 ± 37,2* 895,4 ± 41,6* 645,7 ± 31,2
cGMP (nmol/g liver) L-303 37,5 ± 3,6 16,7 ± 1,15* 22,3 ± 1,84* 38,3 ± 3,5
L-500 37,5 ± 3,6 18,5 ± 1,63* 20,6 ± 1,73* 36,8 ± 4,1
Sum of adenine nucleotides (mcmol/g liver) L-303 4,34 ± 0,08 2,78 ± 0,07* 2,84 ± 0,07* 4,36 ± 0,09
L-500 4,34 ± 0,08 2,54 ± 0,07* 2,77 ± 0,06* 4,34 ± 0,23
Phosphocreatine (mcmol/g liver) L-303 1,27 ± 0,06 0,47 ± 0,03* 0,56 ± 0,04* 1,25 ± 0,08
L-500 1,27 ± 0,06 0,45 ± 0,03* 0,62 ± 0,04* 1,32 ± 0,07
Energy potential : (ATP + 1/2 ADP)/ (ATP + ADP + AMP) L-303 0,66 ± 0,02 0,27 ± 0,03* 0,349 ± 0,02* 0,66 ± 0,03
L-500 0,66 ± 0,02 0,27 ± 0,02* 0,34 ± 0,03* 0,65 ± 0,04
Mg2+- ATP-ase (P mcmol/mg protein 1 h), the mitochondria L-303 81,46 ± 4,7 42,58 ± 3,7* 54,6 ± 3,8* 82,53 ± 5 26
L-500 81,46 ± 4,7 45,3 ± 3,44* 56,23 ± 4,52* 79,6 ± 4,82
Ca2+ ATP-ase (P mcmol/mg protein 1 h), the mitochondria L-303 73,52 ± 5,1 39,65 ± 3,2* 43,76 ± 4,1* 74,37 ± 4 93
L-500 73,52 ± 5,1 38,93 ± 3,56* 48,63 ± 3,74* 71,96 ± 5,43
Note: * — reliable differences with control, p < 0,05.
Statistically significant differences in terms of the control of energy metabolism in the liver, in the experimental and control groups are not established at LD50 1/1000 dose of the given xenobiotics.
The results show that in the animals treated L-303 laproxid at doses of 1/10 and 1/100 of LD50 ATP content in liver has decreased, respectively, by 76.34 % and 68.31%, ADP - 64.07 and 58%, 60%, cGMP - 55.47% and 40.54 %, the amount of adenine nucleotides - 35.95 % and 34.57 %, phosphocreatine - 63% and 55.91 %, the energy potential value of cells has decreased by 59.1% and 47.22% as well as the activity of Mg2+- ATP-ase -47.73 % and 32.98%, Ca2 +- ATP-ase - 46.07 % and 40.48 % on higher levels of cAMP by 110.9 % and 90.24%, an inorganic phosphate - 138.34% and 62%, cAMP - 43.8% and 37.8% compared with the control group.
Under L-500 laproxide influence at doses of 1/10 and 1/100 of LD50, respectively, ATP content has reduced by 78.52% 69.2%, as well as ADP - 68.22% and 60.94%, cGMP - 50.67% and 45.07%, the amount of adenine nucleotides - 41.48% and 36.18%, phosphocreatine -64.57% and 51.19%, value of the energy potential of the cell - 59.1% and 48.49%, the activity of Ca2 +- ATP-ase -47.05% and 33,86%, Mg2 +- ATP-ase - 44.39% and 39.98% on increasing the concentration of cAMP by 101.2% and 92.68%, inorganic phosphate - 155.6% and 64.4%, cAMP - 44.6% and 37.6%.
Conclusions
Under conditions of prolonged influence of the L-303 and L-500 laproxides at sub-toxic doses in white rats
inhibition of bioenergetic processes, the prevalence of the catabolism over restoration synthesis is observed.
2. Investigated xenobiotics at doses of 1/10 and 1/100 of LD50 have the ability to reduce the liver ATP content, ADP, cGMP, adenine nukletidov, creatine phosphate, the value of the energy potential of the cells, they significantly weakened the activity of Mg2 +- ATP-ase , Ca2 +- ATP-ase.
3. Negative effect of the L-303 and L-500 laproxides on macroergic compounds metabolism in the liver is manifested by an increase, compared with the control, of AMP, cAMP, inorganic phosphorus levels.
4. Statistically significant differences have not been established between the results obtained in the experimental groups and control if laproxide toxification was at a dose of 1/1000 of LD50.
5. Imbalance in terms of bioenergy metabolism in albino rats confirms the presence of laproxides hepatotoxicity under influence at the sub-toxic doses leading to further metabolic disturbances.
Prospects for further research
The results can be the basis for further study of the laproxides influence at sub-toxic doses on the hepatocytes mitochondria metabolic state under subacute experiment.
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