COMPARATIVE STUDY ON THE INFLUENCE OF SOME SALICYLIC ACID DERIVATIVES AND SULPHANILAMIDE ADMINISTRATION ON SERUM TRANSAMINASIS
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1 Lucrări ştiinţifice Zootehnie şi Biotehnologii, vol 40(1), (2007) Timişoara. COMPARATIVE STUDY ON THE INFLUENCE OF SOME SALICYLIC ACID DERIVATIVES AND SULPHANILAMIDE ADMINISTRATION ON SERUM TRANSAMINASIS STUDIUL COMPARATIV AL INFLUENTEI ADMINISTRĂRII UNOR DERIVAŢI AI ACIDULUI SALICILIC ŞI A SULFANILAMIDEI ASUPRA TRANSAMINAZELOR SERICE STANA LETIŢIA*, TRIF ALEXANDRA*, PADURE MIRABELA**, GRĂVILĂ CORINA***, MUSELIN F*., IVANCOV ROXANA* *Faculty of Veterinary Medicine Timişoara, România **Industrial Chemistry and Environmental Engineering Faculty *** Faculty of Animal Sciences and biotechnologies, Timişoara, România The experiment has been done to estimate the impact of 5ClSA-SA a new synthesis product (the amide of chlorine salicylic acid with sulphanilamide) comparing to salicylamide and sulphanilamide, basis substances for its synthesis. These three substances have been administrated intraperitoneal to Wistar rats. After the fifth and the seventh administrations have been determined the activities of ALT and AST plasmatic enzymes. ALT and AST transaminasis have increased in all experimental batches, the highest values being recorded for5clsa-sa batch. Some xenobiotics thru the actions induced by the generated free radicals can demonstrate their toxic effect. The increasing of ALT activity has shown the hepatic toxicity. The increasing of AST can be induced by a possible hemolytic effect of studied substance. Key words: rats, chlorine salicylic compounds, transaminasis Introduction The study of the action mechanisms on the exposure to chemical agents, the cause of acute and chronically affect of biological systems has given important data in applied toxicology, providing a series of effects on the lab animals and the possibility of exploitation on human [14]. This experiment was aimed to hacking the inducted effect of an synthesis product administration the amide of 5 chlorsalicylic acid with sulphanilamide and of the substances that stay at the base of the products synthesis, salicilamide and sulphanilamide, on the aminotranspherasis activities. 183
2 Materials and Methods The experiment was done on male Wistar rats, of g weight, grouped in 7 batches: one control (n=7) and six experimental (E 1 and E 4 : 0,44 mg 5ClSA-SA/g. m.c./day; E 2 and E 5 : sulphanilamide 0,23 mg /g. m.c/day; E 3 and E 6 salicilamide: 0,185 mg /g m.c./day). The salicilamide was administrated in 1/10 DL 50, the 5ClSA-SA dose was calculated so that 0.44 mg salicylic compound s nucleus which is included in his structure to be in equal amount with the one contained in the administrated 5ClSA-SA dose. The administration was made intraperitoneal: at E 1, E 2 şi E 3 5 administrations and at E 4, E 5 şi E 6 7 administrations. As for the controls, they have been administrated an equivalent volume of distilled water. Blood samples were taken from the cord on coagulant (EDTA) as follows: at the begging of 3 controls, in 120 hours from the first administration from E 1, E 2 and E 3. After 168 hours the harvesting was made by slaughtering through euthanasia from the E 4, E 5 and E 6 and other 3 controls. Were determinates the transaminasic activities: alanin aminotranspferasys (ALT) and aspartat aminotranspferasys (AST) with Coulter Maxim Beckman Hematology Automatic Analyzer. Results and Discussions The results obtained during this study are presented in table 1 and graphs 1 and 2. Table 1 Mean values of aminotranspherasys AST and ALT activities on control and experimental batches AST ALT 5 administrations 7 administrations C E 1 E 2 E 3 E 4 E 5 E 6 55± 146± 97,5± 127± 176± 118,7± 131,5± 3,11 3,25 6,1 2,33 6,3 4,65 4,4 31± 60,6± 46,1± 46,1± 73,5± 62,7± 69± 4,2 2,3 5,4 3,2 5,5 5,0 2,9 At 120 hours from the administration it can be seen at all the experimental batches the increase of AST values compared to control (E 1 :+165%; E 2 : +77%, and E 3 : + 130%) (table 1) remarked at 168 hours from the beginning of the experiment, the continuous increase of the enzymatic activity compared to control (E 4 : +220%; E 5 :+114%; E 6 : +138%%,). 184
3 The ALT values have also increase compared to control after the 5 th administration (after 120 h) (E 1 :+95,5%; E 2 and E 3 : +48,7%) as well as after 168 hours after the first administration (E 4 : +137%; E 5 :+102,2%; E 6 : +122%). AST values dynamic on control and experimental batches Graph h E1,E4 E2,E5 E3,E6 C ALT values dynamic on control and experimental batches Graph h E1,E4 E2,E5 E3,E6 C From the first and the second graphics it can be observed that the 5ClSA-SA substance has led to the highest values of aminotranspherasis compared to control. The AST aminotranspherasis determinate by the salicilanilide and sulphanilamide 185
4 administration has presented at the end of experiment similar values but lower then those induced by 5ClSA-SA. (-32,95% respectively 25,56%). It was the same in the case of alanin aminotranspherasis which reached lower values then those determined by the synthesis product 5ClSA-SA (-14,7% respectively 6,1%). At control, AST and ALT values were in normal limits after some authors [10, 13] and were different after others [6, 11]. AST : ALT ratio (DeRittis coefficient) calculated for rats, from different literature references for aminotranspherasis [10, 11, 13] is close to the value for humans from the literature (between 1 and 2). Under the action of noxys the ALT activity in serum increase faster which determinate a proper fraction value of the DeRittis coefficient. In hypercritical lesions DeRittis coefficient value increase, an aspect also remarked in this experiment at 120 h when all his values were greater 2. At 168 h although AST values increased at E 5 and E 6 batches thru the increase with 1% of ALT values, DeRittis ratio decrease under value of 2, getting close to control. In case of the batch with 5ClCSA-SA product DeRittis coefficient remained at a value of 2.39 which indicate hepatic processes. Our AST enzyme increase has taken to an DeRittis coefficient of 2,39, which revealed a liver affection determinate either by the administrated substance or by the products of metabolism that can induce toxically effects. Hepatotoxicity was indicated by the AST activity increase from plasma and the liver severs necrosis [4]. Hiperenzyme illustrate the critical action speed when the time for cell adaptation is missing as well as the number of injured hepatocites, but it doesn t imply unfavorable evolution [12]. Sulphanilamide represents the basis nucleus of antibacterial sulphamides. Sulphamide metabolization, especially N 4 -acetilare, is made in liver. The acetylate acids are bacterial inactive but keep the toxicity. Sulphamides produce reverse reactions, can lead to unwanted phenomenon: liver diffuse hepatitis with necrosis, hemolytic anemia and others. Studies regarding the effect of some salicylic acids compounds, among which salicylamide [15], indicate the reduction of erythrocytary membrane fluidity. Its consequence is the modification of proteins proprieties as enzymes or transporters. The increase of aminotranspherasis from serum can be a consequence of toxic effect of drugs. One of the oxidative stress types is the ambient ones. It can be induced by the radiations, drugs, unmedicinal xenobiotics, atmospheric pollutants and so on [2, 3, 4, 7, 8]. The xenobiotics metabolizations, in the system of microsomal oxidative enzymes, can determinate hepatocitary injuries. The experimental and clinical studies prove the fact that the induced tisular aggression from the reactive species of oxygen can be relevant to most of the hepatic affections [2]. 186
5 Conclusions The administrated substances, the amide of 5 chlorsalicylic acid with sulphanilamids, salicylamine and sulphanilamide, have induced the significant increase of AST and ALT aminotranpherasis. The highest effect was determined by the 5ClSA-SA. The increase of aminotranspherasis values can by the consequence of the oxidative stress determined by the toxic effect of drugs. The highest value of DeRittis coefficient was reached at administration of 5ClSA-SA synthesis product during the entire experiment that suggests a possible liver injury provoked by this substance. Bibliography 1. Arduini,A., A.,Peschechera, F., Giannessi, P., Carminati, (2004),Improvement of statin associated myotoxicity by L-carnitine, J.Thromb Haemost,2,pag Dejika,D. (2000) Stresul oxidative în bolile interne, Ed. Casa Cărţii de Ştiinţă Cluj-Napoca. 3. Fadeyi,O.O.,C.,A., Obafemi,C.,O.,Adewunmi, E.,O.,Iwalewa, (2004), Antipyretic, analgesic, anti-inflmmatory and cytotoxic effects of four derivatives of salicylic acid and anthranilic acid in mice and rats,african Journal of Biotechnology, vol.3(8)pag Ghazi,I.M., ZA., Mohamed, SA.,Ismail., A.,Kamal, (1986), Antibacterial effect and toxicity of synthesized salicylanilide derivatives, Zentralbl Mikrobiol, 141(3), pag Glinsukon, T., S., Tazcharpipranai, C., Toskulkao -(1978)-Plasma,GOT, GPT, rats, J.Cellular and Molecular Life Sciences (OMLS), vol.34,no Hiroyuki, A., H., Kodama, N., Matsuoka, I., Yamaguchi, (1997) Stress Increases Plasma Enzyme Activity in Rats: Differential Effects of Adrenergic and Cholinergic Blockades Pharmacology, Japan Vol. 280, Issue 3, Jacshke,H., G.J., Gores, A.I., Cederbaum, J.A.,Hinson, D., Pessayre, J.J,(Lemasters 2001), Mecanism of hepatotoxicity, Toxicological Sciences, 65(2),pag Jarlijeci, Milena, T., M.,Takač D., V., Topič.- ( 2004) FT IR and NMR spectroscopic studies of salicylic acid derivatives.ii.comarision of 2-hydroxyand 2,4-and 2,5-dihydroxy derivatives, Acta Pharm.54, Kamisako,T., Y., Adachi, T.,Yamamoto, (1990)- Effect of UDP glucuronic acid depletion by salicilamide on biliary bilirubin excretion in the rat, J. Pharmacol.Exp.Ther.254, pag Kechrid,Y., S.,Amamra, N., Bouzerna, (2006),The effect of zinc deficiencz on zinc status, carbohzdrate metabolism and Progesterone level in pregnant rats, Turk.J.Med.Sci.36(6), pag
6 11. Kim J.,S., J.,B.,Ju,C.,W.,Choi, S.,C.,Kim, (2006), Hzpoglicemic and Antihyperlipemic effect of four Korean medicinal plants in alloxan induced diabetic rats, American Journal of Biochemistryand biotechnology, 2(4), pag Marin F.(1995)- Explorări clinice şi morfofuncţionale în medicină Ed. Tipomur, Târgu Mureş 13. Nakamura,A., A., Imaizumi, R., Niimi, Z., Zanagawa,T., Kohsaka, E.J.,Johns (2005)Adenoviral deliverz of the b 2 -adrenoceptor gene in sepsis: a subcutaneous approach in rat for kidnez protection, Clinical Science, 109,pag ed STUDIUL COMPARATIV AL INFLUENTEI ADMINISTRĂRII UNOR DERIVAŢI AI ACIDULUI SALICILIC ŞI A SULFANILAMIDEI ASUPRA TRANSAMINAZELOR SERICE STANA LETIŢIA*, TRIF ALEXANDRA*, PADURE MIRABELA** GRĂVILĂ CORINA***, MUSELIN F*., IVANCOV ROXANA* *Faculty of Veterinary Medicine Timisoara, Romania **Industrial Chemistry and Environmental EngineeringFaculty *** Faculty of Animal Sciences and biotechnologies, Timisoara, Romania S-a realizat un experiment în scopul evaluării impactului asupra enzimelor serice ALT şi AST, a unui produs nou de sinteză 5ClSA-SA (amida acidului clorosalicilic cu sulfanilamida) comparativ cu salicilamida şi sulfanilamida, substanţe care stau la baza sintezei acestuia.administrarea celor trei substanţe s-a făcut la şobolani Wistar, intraperitoneal, timp de 7 zile. După 5 şi respectiv 7 administrări, s-au determinat activităţile enzimelor plasmatice ALT şi AST.Transaminazele Alt şi AST au înregistrat creşteri la toate loturile experimentale, cele mai mari valori remarcându-se la lotul cu 5ClSA-SA.Unele xenobiotice pot să-şi exercite efectul toxic prin intermediul recţiilor induse de radicalii liberi generaţi. Hepatotoxicitatea a fost indicată prin creşterea activităţii ALT din plasmş. Creşterea AST poate fi indusă şi de un posibil efect hemolitic al substanţelor studiate. Cuvinte cheie: şobolani, derivaţi ai acidului salicilic, transaminaze 188
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