Lipase Activity in Albino Rats Treated with an Antimalarial Drug

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1 ISSN: Volume 3 Number 11 (November-2015) pp Lipse Activity in Albino Rts Treted with n Antimlril Drug K. N. Agbfor* Deprtment of Biochemistry, Ebonyi Stte University, Abkliki, Nigeri *Corresponding uthor KEYWORDS Pncres, Serum, Mlri, Totl protein, Toxicity A B S T R A C T Mlri is common disese cused by Plsmodium prsites, which infect red blood cells through the bites of n infected femle nopheles mosquito. Anti-mlri drugs hve been reported to show vrious side effects. The present reserch ws set up to exmine the ctivity of lipse in serum of lbino rts s mesure of possible toxic effect of n ntimlril drug. The lbino rts were plced in four (4) groups, A, B, C, nd D, with five rts per group. Groups A, B, nd C were treted with orl doses of 8.0, 16.0, nd 32.0mg/kg body weight respectively of the drug solution, while group D served s the control. The tretment lsted for seven (7) consecutive dys. There ws generl decrese in verge body weight, feed nd wter intke of the groups treted with the drug reltive to the control. The tretment of the nimls did not produce ny significnt difference (P> 0.05) in totl protein concentrtion between test nd control. The lipse ctivity in the serum of the lbino rts in the test groups ws found to be significntly higher (P< 0.05) thn in the control. This effect ws found to increse with the doses. The findings in this reserch re indictive tht n ntimlril drug my be toxic to the pncres. Introduction Mlri is mosquito-borne infectious disese of humns nd other nimls cused by protists ( type of microorgnism) of the genus Plsmodium [1]. It begins with bite from n infected femle Anopheles mosquito, which introduces the protists through sliv into the circultory system. In the blood, the protists trvel to the liver to mture nd reproduce. Mlri cuses symptoms tht typiclly include fever nd hedche, which in severe cses cn progress to com or deth [2]. The disese is widespred in tropicl nd subtropicl regions in brod bnd round the equtor, including much of Sub-Shrn Afric, Asi, nd the Americs [3]. 73

2 Five species of Plsmodium cn infect nd be trnsmitted by humns. The vst mjority of deths re cused by P. flciprum nd P. vivx, while P. ovle, nd P. mlrie cuse generlly milder form of mlri tht is rrely ftl. P. flciprum is the principl prsite in sub-shrn Afric nd ccounts for pproximtely 90% of ll deths worldwide from mlri. Most ftl cses ffect children ged under 5 yers, where levels of cquired immunity to the prsite re often insufficient to llow survivl beyond infncy [4]. The zoonotic species P. knowlesi, prevlent in Southest Asi, cuses mlri in mcques but cn lso cuse severe infections in humns. Mlri is prevlent in tropicl nd subtropicl regions becuse rinfll, wrm tempertures, nd stgnnt wters provide hbitts idel for mosquito lrve. Disese trnsmission cn be reduced by preventing mosquito bites by distribution of mosquito nets nd insect repellents, or with mosquitocontrol mesures such s sprying insecticides nd drining stnding wter. Antimlril drugs re clssified into tree ccording to site of ction in mlri prsites. These re those tht ct on the food vcuole, drugs tht block metbolic synthesis nd oxidtive processes, nd those tht interfere with membrne processes[5]. Knowledge of these sites of ction hs enbled identifiction of new drugs with the most promising potentil for development. Current ntimlril strtegies prioritize combintion therpies such s tovquone/ progunil or rtemether/lumefntrine nd prolonged tretments to limit the risk of inducing drug resistnt Plsmodium [6]. An ntimlril drug contining rthemether nd lumefntine. The vilbility of rtemisinin-bsed compounds, derived from the Chinese plnt Artemisi nnu, during the pst decde hs introduced new er in the tretment of mlri. Effective nd well tolerted, these drugs pper to offer substntil benefits over trditionl gents such s chloroquine or sulfdoxinepyrimethmine. Artemisinin-bsed combintion therpies (ACT) re recommended s first-line therpy for mlri by the World Helth Orgniztion (WHO) nd hve been ccepted s minsty of tretment in more thn 70 countries [7]. The mechnism by which rtemether exerts their ntimlril ctivity remin contentious. Nevertheless, most studies concur tht the ctivity of rtemether nd mny if not ll of its potent derivtives results from reductive scission of the peroxide bridge by reduced heme iron, which is produced inside the highly cidic digestive vcuole s it digests hemoglobin. In support of this, recent study with fluorescent rtemethertrioxone derivtives provided evidence for their rpid ccumultion in the digestive vcuole nd their ctivtion by neutrl lipid-ssocited heme. The selection of rtemether nd lumefntine s combintion is due to their reported synergistic effects ginst P. flciprum in vitro. Lumefntine is lso chosen due to its lrge pprent volume of distribution [8]. Lipses (EC tricylglycerol cylhydrolse) re group of wter soluble enzymes, which exhibit the bility of cting t the interfce between queous nd orgnic phses. They primrily ctlyze the hydrolysis of ester bonds in wter insoluble lipid substrtes. However, some lipses re lso ble to ctlyze the processes of esterifiction, interesterifiction, trnsesterifiction, cidolysis, minolysis nd my show enntio selective properties [9]. Serum lipse elevtions hve been reported to be positively ssocited with correct dignosis of cute pncretitis, with dignostic efficiencies of 94 per cent. A 74

3 close correltion between elevtion of serum lipse hs been observed in both extrpncretic nd pncretic disese processes. Serum lipse is better test thn serum mylse either to exclude or to support dignosis of cute pncretitis [10]. Administrtion of n ntimlril drugis sometimes done with cution due to some reported side effects, such s skin rshes, difficulty in sleeping, nuse, vomitting, dirrhe, nd coughing. Administrtion of n ntimlril drugtblets in high doses cn led to toxicity of the heptocytes [11]. Hence, the present study investigted the effect of orl dministrtion of n ntimlril drugon lipse ctivity s n index of possible pcretic toxicity. Mterils nd Methods Collection of Smples Twenty (20) dult mle lbino rts weighing g were obtined from Zoology Deprtment, University of Nigeri Nsukk, Enugu stte, nd trnsported to the niml house, Deprtment of Biochemistry, Ebonyi Stte, University, Abkliki. Two pckets of An ntimlril drugwere purchsed from Elegnt drug store Abkliki, Ebonyi Stte, Nigeri. Animl (Rts) Hndling nd Tretment Ethicl pprovl for niml use in reserch ws given by Ebonyi Stte University Ethics nd Reserch Committee. The 20 Albino rts were rndomly plced in 4 cges, lbeled (A, B, C nd D), ech contining 5 rts. All the nimls were llowed free ccess to wter nd feed (growers msh), nd were cclimtized for 7 dys before dministrtion of smple commenced. After cclimtiztion, doses of 8, 16, nd 32mg/kg body weight of drugsolution were dministered orlly to rts in groups A, B, nd C respectively for seven consecutive dys. Group D served s control nd received distilled wter. After seven dys of drug dministrtion, the rts were strved overnight, scrificed under mild nesthesi using chloroform, nd blood smples were collected from the lbino rts by crdic puncture. Mesurement of Prmeters Serum ctivity of lipse ws mesured ccording to the method of Yng nd Biggs [12], while Lowry [13] method ws dopted to determine totl protein. Sttisticl Anlysis Dt generted were expressed s men ± SD. Sttisticl significnce of difference ws determined using the progrm SPSS 12 (SPSS, USA) by performing one-wy ANOVA with post-hoc comprisons between the control group nd ech of the treted groups by Ducn s multiple comprison test. A p-vlue less thn 0.05 ws considered sttisticl significnt. Results nd Discussion Tretment of the nimls with solution of the ntimlril drug resulted in decrese in their physicl ctivities nd rte of feed nd wter intke reltive to the control (dt not shown). The ctul biochemicl mechnism behind this observtion is still been investigted. However, it my be due to n upset in generl metbolism cused by the drug. Some mild nd uncommon side effects of rtemether-lumefntrine combintion include loss of ppetite nd wekness [14]. The finding is consistent with the report of Foley nd Tilley [15] on 75

4 tretment of lbino rtscoterm ( combintion of rtemether-lumefntrine). 2, the difference between totl protein concentrtions recorded in the serum of the rts treted with drug solution nd the control ws not significnt (P>0.05). This indictes tht the chemicl constituents of the drug my ply no significnt role in regultion of protein synthesis nd regultion. The verge body weight of the nimls in the groups dministered the drug smple decresed, while tht of the control incresed (tble 1). This result my be ttributed to the reported decresed in feed nd wter consumption. As shown in tble Tble.1 Averge body weight of the rts during seven dys of drug dministrtion Dy of dministrtion BODY WEIGHT (g) A B All vlues re men ± stndrd devition; N = 5 C D Key: Group A = 8.0mg/kg body weight Group B = 16.0mg/kg body weight Group C = 32.0mg/kg body weight Group D = distilled wter Tble.2 Averge serum lipse ctivity nd protein concentrtion fter seven dys of drug dministrtion GROUP A B C D ENZYME ACTIVITY (U/l) b c TOTAL PROTEIN (mg/ml) All vlues re men ± stndrd devition; N = 5. Vlues in the sme column bering different superscripts differ significntly (P<0.05) Key: Group A = 8.0mg/kg body weight Group B = 16.0mg/kg body weight Group C = 32.0mg/kg body weight Group D = distilled wter 76

5 Serum lipse ctivity of the nimls fter seven dys of tretment is presented in tble 2. The ctivity of the enzyme obtined in groups dministered the drug ws significntly higher (P<0.05) thn in the control. The exct mechnism responsible for this elevtion of serum lipse ctivity is subject to further studies. However, it my be s result of dmge to pncretic cells which cuses the enzyme to lek into circultion. Lipse ssy s biomrker of pncretic dmge hs sensitivity nd specificity of 80% nd 60%, respectively [16][17]. The serum concentrtion of lipse increses within 3 6 hours of onset of dmge nd peks within 24 hours [18]. The incresed serum level stys for round 7 14 dys before it comes down to the norml level [19]. In contrst to mylse, lipse is rebsorbed in renl tubules nd stys for long t higher concentrtion, thereby giving greter sensitivity in ptients with delyed presenttion. Pncretic lipse is four times more ctive thn mylse nd it is less ffected by exocrine pncretic deficiency occurring in ptients of chronic pncretitis [20]. Hypertriglyceridemi does not influence the serum lipse ssy s hppens in the cse of serum mylse. Incresed serum level of lipse cn lso be seen in mny intr-bdominl pthologies including cute cholecystitis, ppendicitits, inflmmtory bowel disese, intestinl ischemi, obstruction, perfortion, nd renl insufficiency. According to recent guidelines from UK, serum lipse should be preferred for dignosis of pncretic dmge over serum mylse wherever vilble [21]. The rtemether-lumefntrine combintion is generlly well tolerted. The most frequently reported dverse effects in peditric clinicl trils hve been fever (29%), cough (23%), vomiting (18%), hedche (13%), nd norexi (13%). Dizziness, nuse, bdominl pin, dirrhe, rshes, rthrlgi nd mylgi, stheni, ftigue, nd elevtions in serum trnsminses hve been reported in 1-10% of children [22]. These observed effects of orl consumption of n ntimlril drugon body weight nd serum lipse ctivity were found to be dosedependent. Conclusion From the findings of this reserch, it cn be suggested tht orl dministrtion of n ntimlril drugmy be toxic to the pncres. It my lso ffect ppetite nd physicl helth. However, studies re in progress in our lbortory to estblish these findings, nd identify the possible mechnisms responsible. References 1. Crvlho, L.J., Dniel-Ribeiro, C.T., Goto, H., Mlri vccine: cndidte ntigens,mechnisms, constrints nd prospects. Scnd. J. Immunol. 56: Smith, T.G., Ayi, K., Serghides, L., McAllister, C.D., ndkin, K.C. ( 2002). Innte immunity to mlri cused by Plsmodium flciprum. Clin. Invest. Med. 25, Olumese, P. ( 2005). Epidemiology nd surveillnce: chnging the globl picture of mlri myth or relity? Act Trop. 95, WHO nd UNICEF, World Mlri Report White, N. J. (1997). Assessment of the phrmcodynmic properties of ntimlril drugs in vivo. Antimicrob Agents Chemother,41: White, N. J. nd Pongtvornpinyo, W. (2003). The de-novo selection of drug 77

6 resistnce in mlri prsites.philos Trns R SocLond B BiolSci2003, 270: WHO (2001). Antimlril drug combintion therpy. Report of WHO technicl consulttion. WHO/CDS/RBM/ , Genev, Switzerlnd. 8. Jckson, Y., Chppuis, F., Loutn, L. nd Tylor, W. (2006). Mlri tretment filures fter rtemisininbsed therpy in three exptrites: could improve mnufcturer informtion help to decrese the risk of tretment filure? Mlri Journl, 5: Hsn, F.; Shh, A.; Hmeed, A. Methods for detection nd chrcteriztion of lipses: A comprehensive review. Biotechnol. Adv., 2009, 27, Chse, C. W., Brker, D. E. nd Burns, R. P. (1996). Serum mylse nd lipse in the evlution of cute bdominl pin.am Surg., 62(12): Krueger, K. (2006). Mnipultion of host heptocytes by the mlri prsite for delivery into liver sinusoids. Journl of Medicl Science,313 (5791): Yng, J.S. nd Biggs, H.G. (1971). Rpid, relible method for mesuring serum lipse ctivity. Clin. Chem., 17(6): Lowry, O. H. (1951). Protein mesurement with the Folin phenol regent. J Biol Chem. 193(1): Chinw, J. M. (2013). Adverse Drug Event From Artemether/lumefntrine Ingestion: Cse Series, Niger J Ped, 40 (4): Foley, M. nd Tilley, L. (2000). Quinoline ntimlril. Mechnism of ction nd resistnce nd prospect for new gents: Phrmcology nd therpeutics. 7(5): Chng, J. W. Y. nd Chung, C. H. (2011). Dignosing cute pncretitis: mylse or lipse? Hong Kong Journl of Emergency Medicine, 18(1): Lott, J. A., Ptel, S. T., Swhney, A. K., Kzmierczk, S. C. nd Love Jr., J. E. (1986). Assys of serum lipse: nlyticl nd clinicl considertions, Clinicl Chemistry, 32(7): Lippi, G., Vlentino, M. nd Cervellin, G. (2012). Lbortory dignosis of cute pncretitis: in serch of the Holy Gril, Criticl Reviews in Clinicl Lbortory Sciences, 49(1): Mtull, W, R., Pereir, S. P. nd O'Donohue, J. W. (2006). Biochemicl mrkers of cute pncretitis, Journl of Clinicl Pthology, 59( 4): Tietz, N. W. nd Shuey, D. F. (1993). Lipse in serum: the elusive enzyme: n overview, Clinicl Chemistry, 39(5): Werner, J., Feuerbch, S., Uhl, W. nd Büchler, M. W. (2005). Mngement of cute pncretitis: from surgery to interventionl intensive cre, Gut, 54(3): vnagtmel, M., Bouchud, O., Mlvy, D., Delmont, J., Dnis, M., Brette, S., Grs, C., Bernrd, J., Touze, J. E., Gthmnn, I. nd Mull, R. (1999). The comprtive efficcy nd tolerbility of CGP (rtemether +lumefntrine) versus hlofntrine in the tretment of uncomplicted flciprum mlri in trvellers returningfrom the Tropics to The Netherlnds nd Frnce.Int J Antimicrob Agents, 12: