International Journal of Current Research in Biosciences and Plant Biology ISSN: Volume 2 Number 6 (June-2015) pp

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1 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): Originl Reserh Artile Interntionl Journl of Current Reserh in Biosienes n Plnt Biology ISSN: Volume 2 Numer 6 (June-2015) pp Evlution of Heptoprotetive Effet of Hert of Plm (Plmito) Extrt ginst CCl 4 Inue Heptotoxiity in Ault Mle Rts S.Z.M. Al-Ahi* n S.M.Y. Al-Gorny Deprtment of Chemistry, College of Siene, University of Mosul, Irq *Corresponing uthor. A s t r t The im of the present investigtion ws to exmine the effets of hert of plm (plmito) queous extrt in heptotoxiity inue y ron tetrhlorie (CCl 4 ) in Alino ult mle rts. Thirty rts were use n ivie rnomly into five groups, eh of whih ontin six nimls, group І ws serve s negtive ontrol, reeive no tretment, the group ІІ ws trete with orl CCl 4 t ose of 0.5 ml/kg oy weight to inue heptotoxiity every other y for seven y, group ІІІ, ІV n V reeive CCl 4 in similr mnner with queous extrt of plmito ily intke t ifferent oses (100,250,500 mg/kg) respetively for two weeks. Bloo smples otine from oritl sinus punture y use speil pillry tue t pre-tretment n t the en experiment for iohemil nlyses : tivities of liver funtions enzymes were ssesse y estimting lnine mino trnsferse (ALT), sprtte mino trnsferse (AST), lkline phosphtse (ALP), totl, iret n iniret iliruin (TSB, DB n IDB respetively), totl protein (TSP), lumin n totl lipi. The result inite tht plmito hs the ility to protet hepti tissue ginst the toxiity inue y CCl 4 revele through signifint reution of serum tivities of liver enzymes n in the onentrtions of TSB, DB n totl lipi, n with signifint inrese of serum onentrtions of totl protein n lumin. In onlusion, plmito n e propose to protet the liver tissue ginst CCl 4 -inue liver mge in rts. K e y w o r s Antioxint CCl 4 Heptoprotetive Liver Plmito Introution The sttus of herl meiine hs een fst gining groun ll over the worl uring the lst few ees, minly ue to the generl elief tht herl rugs re without ny sie effets esies eing hep n lolly ville (Nith et l., 2011; Kumr et l., 2012). The plmito, known s "hert of plm", is the entrl prt or hert of some wil vrieties of plm trees, it ' s one of suh plnt grown in mny lolities in S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

2 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): ri n semi-ri regions of the worl (Aohtem et l., 2011) is onsiere s goo soure of energy, it ontin vitmins, group of elements like phosphorus, iron, potssium, n signifint mount of lium (El- Gzzr et l., 2009). The informtion rue in the pst four ees suggest tht tes possess iverse meiinl uses inluing ntihyperlipiemi, ntiner, gstroprotetive, nephroprotetive, ntiinflmmtory, n heptoprotetive effets (Blig et l., 2011). It is lso use in the tretment for sore throt, ystitis gonorrhe, n eem n to ountert lohol intoxition (Al-Dihn n Bht, 2012). Besies fruit, the te plm over the enturies hs lso provie lrge numer of other prouts whih hve een extensively use y mn in ll spets of ily life (Agoujil et l., 2011). Euterpe eulis (known s Plmetto) is the most importnt wiely use speies whih lolly (Shimizu et l., 2011). Liver isese is n ute or hroni mge to the liver, usully use y infetion, injury, exposure to rugs or toxi ompouns, n utoimmune proess, or y geneti efet. Chemil suh s ron tetrhlorie (CCl 4 ) tolism rils inue lipi peroxition, mge the memrnes of liver ells n orgnelles, n uses the swelling n nerosis of heptoytes n result to the relese of ytosoli enzymes suh s AST, ALT n ALP into the irulting loo. These enzymes re use s the initors of hemilly inue liver mge (Drotmn n Lwhn, 1978). The CCl 4 is one of the most ommonly use heptotoxins in the experimentl stuy of liver iseses. The heptotoxi effet of CCl 4 is lrgely ue to its tive metolite, trihloromethyl ril umultion in the liver (Thngkrishnkumri et l., 2012). The present stuy ws esigne to elevte the possile heptoprotetive effet of ifferent ose of Euterpe eulis ginst CCl 4 inue liver mge in experimentl rts. Mterils n methos Colletion n uthentition of plnt For the present work the fresh plnt mterils of Euterpe eulis were ollete from Bʼqu ity/dyl /Irq. The plnt ws positively ientifie y Biologist literture in Biologil eprtment /College of Siene /University of Mosul / Irq. The plnt ws wshe with tp wter n rie in she t room temperture, n then it ut into smll prts. Preprtion of Euterpe eulis extrt A out (300gm) of rie plnt were e to (900 ml) of istille wter (1/3, w/v) then rushe y lener t room temperture for overnight using shker. After tht, the mixture ws filtrte through glss woos. The extrt ws sujete to freezing t (- 20 C ) n thw yle for perio of three ys (Kreer et l., 2004), then ultrsoni system ws use to rekge ells wlls (Royt n White, 1987). The mixture were ollete n onentrte to suspene solution uner reue pressure using Lypholyzer system. Experimentl esign Helthy mle lino rts weighting etween ( g) were otine from the niml house in the meiine fulty / Mosul University / Irq. Rts were house uner stnr lortory onition, light n rk yles of 12h, in polypropylene ges n llowe free ess to fee n tp wter uner stritly ontrolle pthogen free onitions with room temperture (25±2ºC), humiity (50±5%).The nimls were rnomly ivie into five groups, omprising of six rts in eh. Group I: Reeive (0.5ml/kg) oy weight of istille wter orlly for 2 weeks n use s the norml ontrol. The nimls in ll groups were kille y nestheti ether on the y 15. Group II: Reeive (0.5ml of mixture of 1:1 V/V in orn oil/kg) oy weight CCl 4 orlly y gvges tue to ll groups exept norml ontrol group to inue heptotoxiity on 1 st, 3 e, 5 th n 7 th ys of experiment (Kpur et l., 1994). Group III, VI n V reeive CCl 4 in similr s group II n they were trete ily with ifferent oses of rue ol queous extrt of Euterpe eulis. Group III: Reeive (0.5ml/kg) oy weight ru ol queous extrt of Euterpe eulis ily orl intke t the ose of (100mg/kg) for 14 ys. Group IV: Reeive (0.5ml/kg) oy weight ru ol queous extrt of Euterpe eulis ily orl intke t the ose of (250mg/kg) for14 ys. S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

3 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): Group V: Reeive (0.5ml/kg) oy weight ru ol queous extrt of Euterpe eulis ily orl intke t the ose of (500mg/kg) for 14 ys. Bloo smples from ll groups were ollete weekly from Oritl sinus punture y use speil pillry tues, left stn in serum tues for (30 minutes) to e ogulte. Serum smples were ollete y entrifugtion t (3000 rpm) for (20 minutes) t room temperture. The ler, non-hemolyse ser ws seprte n store t (- 20 C ) for mesurements of iohemil nlyses of liver funtions (Att et l., 1983). Biohemil nlyses The AST n ALT tivities were mesure spetrophotometrilly y using the metho of Reitmn n Frnkle (Anerson n Cokyne, 1993). ALP ws mesure y the metho evise y Bowers n MCom, whih llows lultion of ALP tivity sing on the molr sorptive of P- nitrophenol (Tietz, 1999). Totl serum protein (Bishop et l., 2005) n serum lumin ws etermine quntitively y olorimetri metho use romoresol green (Doums et l., 1971).The totl proteins minus the lumin gives the gloulin. The iliruin n onjugte iliruin were etermine s esrie y (Bishop et l., 2005). The unonjute iliruin onentrtions were lulte s the ifferene etween totl n onjute iliruin onentrtions. The totl lipi etermine y hete the serum with onentrte sulphuri i, then ret the mixture with phosphovniline regent (Toro n Akermnn, 1975). Sttistil nlysis The t presente in this investigtion were expresse s the men ± Stnr error (men ± SEM) of the men. The signifint ifferene mong the mens hs een sttistilly nlyze y one-wy nlysis of vrine ANOVA, p<0.05 ws onsiere s sttistil signifine using SPSS Softwre (Sneeor n Cohrn, 1986). Results n isussion Effet of plmito queous extrt on oy weight of ontrol n in CCl 4 inue hepti mge in rts No nimls ie uring the stuy n no linil n ehviorl hnges were oserve in ny of the nimls fter ministrtion of oth CCl 4 intoxite n the plmito extrt. Fifteen ys fter tretment with plmito extrt, there were signifint inrese in the weights of oies in the four trete-groups of rts ompre to those in the ontrol group espeilly in group II in the first n seon weeks s shown in Tle 1. Tle 1. Effet of plmito queous extrt on oy weight of ontrol n in CCl 4 inue hepti mge in rts. Boy weight (gm) Tretment (men ± SEM) 1 st week 2 n week Group I 183.2± ±26.40 Group II 320.9± ±41.81 Group III 257.9± ±20.39 Group VI 245.1± ±21.67 Group V 265.5± ±15.85 Vlues with ifferent letters within olumn iffer signifintly t (p<0.05), group II ompre with group I n groups II, VI n V were ompre with group II. Group I = Control group, Group II = CCl 4 -trete group, Group III=Animls trete with CCl 4 intoxition prior plmito extrt t ose (100 mg/kg), Group VI = Animls trete with CCl 4 intoxition prior plmito extrt t ose (250 mg/kg), Group V = Animls trete with CCl 4 intoxition prior plmito extrt t ose (500 mg/kg). Effet of plmito queous extrt on enzymes tivity of serum AST, ALT n ALP in norml n CCl 4 inue hepti mge in rts The effets of extrt t three ose levels (100, 250 n 500 mg/kg) on serum mrker enzymes in CCl 4 - inue hepti injury re shown in Tle 2. Hepti injury inue y CCl 4 use signifint rise (p<0.05) in mrker enzymes AST, ALT n ALP in CCl 4 intoxite group (group II) in omprison with ontrol group (group I). Aministrtion of plmito extrt t three ifferent ose levels ttenute the S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

4 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): inrese levels of the serum enzymes, proue y CCl 4, for oth two weeks n use susequent reovery towrs normliztion espeilly t the seon week. Cron tetrhlorie (CCl 4 ) is ommonly use to inue liver mge tht histologil resemles virl heptitis. Toxiity egins with the hnge in enoplsmi lote in the intrellulr strutures. The toxi metoli CCl 3 ril (. CCl 3 ) is proue n further rets with oxygen to give trihloromethyl peroxy ril. Cytohrome P450 is the enzyme responsile for this onversion. The most serious onsequene of free ril-inue liver injury is lipi peroxition, n it hs een foun tht, free ril n use oxitive mge to ellulr proteins n lter ellulr funtion (Johnston n Kroening, 1998; Thngkrishnkumri et l., 2012). This les to the pthologil hnges suh s epression of protein synthesis, elevtion levels of serum mrker enzymes. The iohemil evienes presente in this stuy lerly emonstrte the stte of oxitive stressinue hepti tissue mge y CCl 4 tretment, mnifeste y the lekges of ALT, AST, n ALP in serum (Okuno, 1986). Therefore, the results of group II h signifint elevtion in the tivities of ALT, AST n ALP ompre to the group I, this my return to the tretment with CCl 4, whih is onsiere s potent heptotoxi gent, n these enzymes re highly sensitive to liver injury n they relese in signifint quntities in the serum fter ny oxitive stress in the liver (Uskokovi- Mrovi et l., 2007). Serum ALP lso inrese signifintly for the heptoellulr injury n holestsis so lrge quntities of this enzyme were relese. These results re in greement with (Pri n Krthikesn, 2007; A n Al-Bghi, 2009) whih inite the inrese permeility, mge, n nerosis of heptoytes. In nother sie the results of the groups III, VI n V revele tht the vlues of serum ALT, AST n ALP erese signifintly ompre with group II. Therefore, the reution in tivities of the enzymes y plmito extrt is n inition of stiliztion of serum memrne s well s repir of hepti tissue mge use y CCl 4. Serum levels of trnsminses whih re sensitive initors of liverell injury n re helpful in reognizing heptoellulr iseses. These enzymes return to norml with the heling of hepti prenhym n regenertion of heptoytes (Frnk et l., 2012). The presene of funtionl ntioxint ompouns (suh s selenium, polyphenols, phytohormones n rotenois) in the te plm fruit in the "Rut" n "Tmr" stges might e responsile for lowering the vlues of hepti enzymes tivity (Al-Syye et l., 2013). In ition (Sfi et l., 2010) foun signifint inrese in the hepti tivity of these enzymes fter tretment of ult mle rts with n queous extrt from te plm fruit (4 ml/kg oy weight) for 2 months. One stuy foun tht feeing mixture of te plm fruit to ult mle rts trete with le ette (to inue lipi peroxition) for 7 weeks enhne the tivity of serum glutthione-strnsferse tivity (Al-Syye et l., 2013). Effet of plmito queous extrt on serum totl protein (TSP), lumin n gloulin in norml n CCl 4 inue hepti mge in rts The totl protein, lumin n gloulin onentrtions were signifintly erese t (p<0.05) in CCl 4 intoxite rts (group II) when ompre with ontrol (group I). The effets of extrt t the ifferent oses on CCl 4 -inue hepti injury re shown in Tle 3, whih note tht the (100, 250 n 500 mg/kg) oy weight of plmito extrt (groups III, VI n V) gives signifint erese when ompre with ontrol group while gives signifint inrese in omprison with group II for first n seon week (Tle 3). Protein metolism is mjor projet of liver n helthy funtioning liver is require for the synthesis of the serum proteins, hypoproteinemi is feture of liver mge ue to signifint fll in hroni liver isese (Hussein et l., 2007). The reution in the level of TSP, lumin n gloulin in the group II inite the effet of CCl 4 on liver funtions. The hypoproteinemi whih ws oserve fter CCl 4 ingestion ut the tren turns towrs norml fter plmito extrt tretment whih ws signifint elevtion in the level of TSP, lumin n gloulin in ifferent oses of plmito extrt groups for oth first n seon week, this my e ue to tretment with the extrt whih preserve the struturl integrity of the liver from the toxi effets, so the signifint hnges in these prmeters inite ntiheptotoxi effet n the ility of plmito extrt to stimulte the regenertion of hepti tissue whih inrese protein synthesis in mge liver n improve the funtionl sttus of the liver ells (Thngkrishnkumri et l., 2012). S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

5 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): Tle 2. Effet of plmito queous extrt on enzymes tivity of serum AST, ALT n ALP in norml n CCl 4 inue hepti mge in rts. Tretment ALT)U/L) (men ± SEM) AST )U/L) (men ± SEM) ALP )U/L) (men ± SEM) 1 st week 2 n week 1 st week 2 n week 1 st week 2 n week Group I 346.2± ± ± ± ± ±11.28 Group II Group III Group IV Group V 493.1± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±7.08 Vlues with ifferent letters within olumn iffer signifintly t (p<0.05), group II ompre with group I n groups II, VI n V were ompre with group II. Group I = Control group, Group II = CCl 4 -trete group, Group III = Animls trete with CCl 4 intoxition prior plmito extrt t ose (100 mg/kg), Group VI = Animls trete with CCl 4 intoxition prior plmito extrt t ose (250 mg/kg), Group V = Animls trete with CCl 4 intoxition prior plmito extrt t ose (500 mg/kg). Tle -3 Effet of plmito queous extrt of on serum totl protein (STP), lumin n gloulins in norml n CCl 4 inue hepti mge in rt. Tretment Group I Group II Group III Group IV Group V STP ( g/l) (men ± SEM) 1 st week 2 n week 9.02± ± ± ± ± ± ± ± ± ±0.15 Alumin ( g/l) (men ± SEM) 1 st week 2 n week 2.80± ± ± ± ± ± ± ± ± ±0.09 Gloulin(g/L) (men ± SEM) 1 st week 2 n week 6.89± ± ± ± ± ± ± ± ± ±0.14 Vlues with ifferent letters within olumn iffer signifintly t (p<0.05), group II ompre with group I n groups II, VI n V were ompre with group II. Group I = Control group, Group II = CCl 4 -trete group, Group III = Animls trete with CCl 4 intoxition prior plmito extrt t ose (100 mg/kg), Group VI = Animls trete with CCl 4 intoxition prior plmito extrt t ose (250 mg/kg), Group V = Animls trete with CCl 4 intoxition prior plmito extrt t ose (500 mg/kg). Effet of plmito queous extrt on serum totl iliruin (TSB), iret iliruin (DB), n in iret iliruin (IDB) in norml n CCl 4 inue hepti mge in rts The results revele tht group II h signifint elevtion (p<0.05) in the levels of serum TSB, DB n IDB ompre to the group I n eh of the groups III, VI n V. In nother sie the results in these groups whih hve ifferent oses of plmito extrt (groups III, VI n V) revele tht the vlues of TSB, DB n IDB erese signifintly in omprison with the group II for oth first n seon week, s shown in Tle 4. Serum iliruin is onsiere s one of the true test of liver funtions sine it reflets the ility of the liver to tke up n proess iliruin into ile. Elevte levels my inite severl illnesses. Hyperiliruinemi my result from the proution of more iliruin thn the liver n proess, mge S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

6 Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6): to the liver impiring its ility to exrete norml mount of iliruin or ostrution of exretory uts of the liver (Olleye et l., 2010). High signifint levels of TSB in CCl 4 trete rts my e ue to rpily proue lipi peroxition of heptoytes memrnes y the tion of free rils (Lee et l., 2004). This my hve resulte in hyperiliruinemi. The signifint reution in the level of totl iliruin in the serum of plmito extrt trete rts suggeste the heptoprotetive potentil of Euterpe eulis extrt ginst CCl 4 intoxition. Effet of plmito queous extrt on serum totl lipi in norml n CCl 4 inue hepti mge in rts The serum totl lipi onentrtion of the experimentl groups whih reeive (100, 250 n 500 mg/kg ) oy weight of plmito extrt were signifintly inrese ompre with the ontrol group (p<0.05), while the vlue of serum totl lipi in these groups erese signifintly in omprison with the CCl 4 inue hepti mge group whih re shown in Tle 5. Tle 4. Effet of plmito queous extrt on serum totl iliruin (STB), iret iliruin (DB) n iniret iliruin (IDB) in norml n CCL 4 inue hepti mge in rts. Tretment STB (µ mol/l) (men ± SEM) DB (µ mol/l) (men ± SEM) IDB (µ mol/l) (men ± SEM) 1 st week 2 n week 1 st week 2 n week 1 st week 2 n week Group I 6.34± ± ± ± ± ±0.84 Group II 68.85± ± ± ± ± ±0.85 Group III 12.14± ± ± ± ± ±0.78 Group VI 18.76± ± ± ± ± ±0.81 Group V 20.31± ± ± ± ± ±0.91 A Vlues with ifferent letters within olumn iffer signifintly t (p<0.05), group II ompre with group I n groups II, VI n V were ompre with group II. Group I = Control group, Group II = CCl 4 -trete group, Group III = Animls trete with CCl 4 intoxition prior plmito extrt t ose (100 mg/kg), Group VI = Animls trete with CCl 4 intoxition prior plmito extrt t ose (250 mg/kg), Group V = Animls trete with CCl 4 intoxition prior plmito extrt t ose (500 mg/kg). Tle 5. Effet of plmito queous extrt on serum totl lipi in norml n in CCl 4 inue hepti mge in rts. Totl lipi (mg/l) Tretment (men ± SEM) 1 st week 2 n week Group I 268.2± ±7.24 Group II 504.8± ±13.37 Group III 315.7± ±37.5 Group V I 434.5± ±22.8 Group V 323.1± ±16.14 Vlues with ifferent letters within olumn iffer signifintly t (p<0.05), group II ompre with group I n groups II, VI n V were ompre with group II. Group I = Control group, Group II = CCl 4 -trete group, Group III=Animls trete with CCl 4 intoxition prior plmito extrt t ose (100 mg/kg), Group VI = Animls trete with CCl 4 intoxition prior plmito extrt t ose (250 mg/kg), Group V = Animls trete with CCl 4 intoxition prior plmito extrt t ose (500 mg/kg). The totl lipi onentrtion ws signifintly inrese in CCl 4 group when ompre with ontrol group, while it ws signifintly erese in groups III, VI n V when ompre with group II fter two weeks of tretments. This fining oinie with the report of (Bhthen et l., 2002; Auelgssim, 2010). Although, (Chir et l., 2007) reporte tht flesh n pit extrts of te plm fruit hve free ril svenging tivities, however, the signifint effet fter two weeks of plmito extrt on serum totl lipi S.Z.M. Al-Ahi n S.M.Y. Al-Gorny (2015) / Int. J. Curr. Res. Biosi. Plnt Biol. 2015, 2(6):

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