The Enzyme Membrain-Associated Complex Activity in Rat Brain under Imidazolin Containing Organic Compounds Action

The aim of the study was to determine the Na + , K + -ATPase, adenylyland guanylyl cyclase synaptosome faction activity in the brain of rats at 30day toxicity by imidazoline mixtures in 1/10 and 1/100 LD 50 doses. Materials and methods. The paper uses examples of MIM with alkyl radicals C 7-9 (MIM7-9) and C 9-15 (MIM9-15). Experiments carried out on the mature male rats Wistar, weighing 180-220 g. Maintenance and manipulation with animals were carried out according to the principles of bioethics. They were subjected to oral intoxification by probe using by aqueous solutions of compounds once daily per 30 days at a 1/100 LD 50 dose. Middle-lethal doses (LD 50 ) were: for MIM79 – 1,8 g/kg; MIM9-15 – 5,0 g/kg of body weight. The animals of the control group were injected by the corresponding volume of drinking water. The parameter study performed at 30 days after the start of the experiment. Each group contained 15 animals. Slaughter was performed by decapitation, with pre-anesthesia by sodium thiopental. The Na + K + -ATP-ase, adenylyl and guanylyl cyclase activity in synaptosomal fraction of rat brain at 30th day of industrial chemical environmental pollutants – imidasoline mixtures influence have been studied in the paper to be necessary for full disclosure of biochemical mechanisms of membranotropic action. Imidasoline-containing mixtures with alkyl radicals С7.9 і С9-15 at 1/10 and 1/100 LD50 doses cause the change of membrane-associated enzyme activity to be confirmed by the decrease of Na + , K + -ATP-ase, adenylyl cyclase activity on the background of guanylyl cyclase increase. Revealed violations are one of pathogenetic links of biochemical mechanisms of imidasoline mixture membranotropic action to be considered in means of correction developing. Summarizing the results, it should be made the following conclusions. Against the background of prolonged toxicity by MIM in 1/10 and 1/100 LD50 doses synaptosome fraction of rat brain membrane enzyme activity is changing, as evidenced by decreased activity of Na + , K + -ATPase, adenylyl cyclase on the background of increasing guanylyl cyclase. Change of membrane-associated enzyme complexes activity in the brain of rats is one of the pathogenetic mechanism of links membrane tropic MIM action to be considered when developing the means of correction.

Inroduction.Actual problem of modern medical biochemistry is a comprehensive studying of the mechanisms of xenobiotic action on humans and means of correction development [7,5].Common xenobiotics able adversely affect health, include a imidazoline mixture (MІМ).This is associated with large volumes of chemicals, widely used in various sectors of the economy (as the basis for industrial production of detergents, antistatic, anticorrosive agents, adhesive additives, etc.), revenues of the sources of drinking water [1,4,14].The clinical picture of rata cute poisoning by MIM is proved to be the symptoms of the central nervous system disorders in the foreground [14].Furthermore, electron microscopy studies indicate significant changes architectonics of the nervous tissue, including violation of the integrity of neurons under conditions of prolonged rat intoxication by MIM [14].Lack of information about the biochemical mechanisms of MIM action on the body encourages them to conduct a comprehensive study, starting primarily with state of the central nervous system.Given the physical and chemical properties of MIM components (the presence of hydrophobic and hydrophilic groups, capacity for chemical reactions to form biologically active compounds, etc.), it is reasonable to research their influence on some characteristics of cell membranes, including membrane-associated enzyme complexes.
The aim of the study was to determine the Na + , K + -ATPase, adenylyl-and guanylyl cyclase synaptosome faction activity in the brain of rats at 30-day toxicity by imidazoline mixtures in 1/10 and 1/100 LD 50 doses.
Materials and methods.The paper uses examples of MIM with alkyl radicals C 7-9 (MIM7-9) and C 9-15 (MIM9-15).Experiments carried out on the mature male rats Wistar, weighing 180-220 g.Maintenance and manipulation with animals were carried out according to the principles of bioethics.They were subjected to oral intoxification by probe using by aqueous solutions of compounds once daily per 30 days at a 1/100 LD 50 dose.Middle-lethal doses (LD 50 ) were: for MIM7-9 -1,8 g/kg; Медичні науки MIM9-15 -5,0 g/kg of body weight.The animals of the control group were injected by the corresponding volume of drinking water.The parameter study performed at 30 days after the start of the experiment.Each group contained 15 animals.Slaughter was performed by decapitation, with pre-anesthesia by sodium thiopental.
Statistical analysis was performed using the computer application package for the processing of statistical information Statistica 6. 1 (StatSoft, Inc., USA).In the case of the normal distribution of data parametric characteristics -the average indicator (M) and standard deviation (s) were used; in its absence nonparametric At the 30th day of observation in synaptosome fraction of rat brain statistically significant (p < 0,001) compared with the control reducing the activity of adenylyl cyclase (AC) was also found (table 2).1/10 LD 50 dose thus was more toxic than 1/100 LD 50 .MIM7-9 and MIM9-15 in 1/10 LD 50 dose reduced AC activity in relation to the control, respectively 58 and 50 %, and in a 1/100 LD 50 dose -41 and 26 %.On the background these changes, there was a statistically significant increased activity of guanylate cyclase (GC).The most expressive effect (p < 0,001) at the same time MIM7-9 commit: on 88 % (in the case of 1/10 LD 50 ) and 66 % (in the case of 1/100 LD 50 ).For MIM9-15 in 1/10 LD 50 dose increase (p≤0,002) GC activity was 54 %, and in a dose 1/100 LD50 -32 %.
Thus, studies have found changes in the activity of membrane-associated enzyme complexes in the brain of rats under prolonged exposure by MIM.The main reasons of this can be a direct effect on enzyme complexes or structural changes in membranes (especially due to free radical processes).Previous experiments have demonstrated that MIM 1/10 and 1/100 LD 50 doses at 30th day actions cause the ratio of plasma membrane phospholipid fractions changes significantly increasing the percentage of phospholipids lysoform [10] and increase microviscosity of lipid phase [9],  Note: p -level of significance compared with control.
-median (Me) and interquartile scope.The critical level of significance during testing statistical hypotheses was accepted p < 0,05.Results and discussion.The groups of rats undergoes oral administration per 30 days by MIM in 1/10 LD 50 dose observed statistically significant (р≤0,005) compared with the control decrease Na + , K + -ATPase activity of synaptosome fraction of the brain by an average of 37 % (table 1).Prolonged exposure by MIM7-9 in 1/100 LD 50 dose was also accompanied by statistically significant relative to the control group of animals, decrease enzyme activity, but less pronounced (at 16 %) than the 1/10 LD 50 dose.Comparison between a distribution rate in the group of animals administered by MIM9-15 in a 1/100 LD 50 dose, and the control did not show any statistically significant difference (p = 0,065).

Медичні науки
which naturally leads, according to many experimental studies [8,11], to reduce the activity of membrane associated enzymes.
The composition of the membranes of all cells as integral protein that converts the energy of ATP to energy gradient of monovalent ions is an enzyme Na + , K + -ATPase.It should be noted that this enzyme is involved into numerous cellular functions and processes associated with ion gradients, particularly in providing the electrical excitability of nervous tissue [3].Therefore, changes in the activity of this enzyme by MIM actions could become an essential reason for their violation.Reducing adenylyl cyclase and increase guanylyl cyclase activity are also a consequence of studied membrane tropic of MIM.As you, The elements of adenylyl cyclase and guanylyl cyclase complexes are known to be integral membrane proteins that's why AC and GC activity are most dependent on the cell membrane [12].In addition, GC can be activated by free radicals, lipid peroxides, which increased formation is observed in the organism of experimental animals under prolonged exposure by MIM [4].
Summarizing the results, it should be made the following conclusions.
1. Against the background of prolonged toxicity by MIM in 1/10 and 1/100 LD 50 doses synaptosome fraction of rat brain membrane enzyme activity is changing, as evidenced by decreased activity of Na + , K + -ATPase, adenylyl cyclase on the background of increasing guanylyl cyclase.
2. Change of membrane-associated enzyme complexes activity in the brain of rats is one of the pathogenetic mechanism of links membrane tropic MIM action to be considered when developing the means of correction.
Perspectives further research.In future the complex research conduction is planned aimed to study the biochemical mechanisms of MIM, including evaluation of antiradical activity and antyperoxid protection.