Original Article IJT 2016; INTRODUCTION. Amadu Kayode Salau*1, Musa Toyin Yakubu 2, Adenike Temidayo Oladiji2

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1 rnin Journl of Toxiology ol 10, No 1, Jn-Fe 2016 Originl Artile Effets of Aqueous Root Brk Extrts of Anogeissus leiorpus (DC) Guill & Perr n Terminli viennioies Guill & Perr on Reox n Hemtologil Prmeters of Diethylnitrosmine-Aministere Rts Amu Kyoe Slu*1, Mus Toyin Ykuu 2, Aenike Temiyo Oliji2 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 Reeive: Aepte: ABSTRACT Bkgroun: This stuy investigte the protetive effets of queous extrts of Anogeissus leiorpus (DC) Guill & Perr (Comretee) n Terminli viennioies Guill & Perr (Comretee) root rks, s well s their 1:1 (w/w) mixture on liver reox n hemtologil prmeters of iethylnitrosmine-trete rts. Methos: Rts were orlly ministere istille wter, iethylnitrosmine (30 mg/kg oy weight one week on weeks 3 n 4), urumin (200 mg/kg oy weight), extrts n 1:1 mixture (200, 400 n 800 mg/kg oy weight) for 4 weeks. Mlonilehye, mrkers of oxitive stress n hemtologil inies were evlute. Results: The extrts n their mixture signifintly (P<0.05) reverse the iethylnitrosmineinue ltertions in the levels of liver mlonilehye, superoxie ismutse, tlse, glutthione peroxise, glutthione reutse, gluose 6-phosphte ehyrogense, glutthione, vitmin C n pltelet ounts. The other hemtologil prmeters (re loo ell ount, hemogloin onentrtion, pke ell volume, men orpusulr volume, men orpusulr hemogloin, men orpusulr hemogloin onentrtion, white loo ell ount, lymphoyte ount n neutrophil ount) were not ffete y iethylnitrosmine n extrts. Conlusion: The extrts possess ntioxint, heptoprotetive n hemoprotetive tivities tht ompre well with urumin. These tivities were etter exhiite y the mixture thn the iniviul extrts. Keywors: Antioxints, Comretee, Curumin, Diethylnitrosmine, Oxition-reution, Oxitive stress. JT 2016; NTRODUCTON rious hemil gents, whether omesti or inustril, to whih mn is expose hve eleterious effets on mn. Suh effets oul e lol, using mge to prt of the oy, or systemi, mging vrious prts of the oy. Exposure to hemils oul lso e from inustril effluents, vehiulr exhusts, n pollution from urning mterils. One of the mny hemils tht mn is expose to is iethylnitrosmine. Diethylnitrosmine (DEN) or N-nitroso iethylmine (NDEA) is n N-nitroso lkyl ompoun, potent heptotoxin n heptorinogen in experimentl nimls, whih uses severe hepti injury [1, 2]. t ours wiely in the environment, is forme enogenously from the intertion of ingeste nitrte or nitrite with seonry mines, n is forme in foo prouts [3]. DEN is foun in too smoke, wter, griulturl hemils, osmetis, phrmeutil prouts n wie vriety of foos like heese, soyen, smoke, slte n rie fish, ure met n loholi everges [2, 4]. t istorts the ellulr ntioxint system n uses lipi peroxition [2, 5]. One of the wys of preventing or reversing suh oxitive mge is using ntioxint gents, whih re present in wie vriety of plnts suh s Anogeissus leiorpus n Terminli viennioies oth of the Comretee fmily. A. leiorpus is known s hewing stik tree (English) n Ayin (Yoru, southwestern 1. Deprtment of Chemil Sienes, Fountin University, Osogo, Nigeri. 2. Deprtment of Biohemistry, University of lorin, Nigeri. *Corresponing Author: E-mil: k.slueen@gmil.om

2 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 rnin Journl of Toxiology Nigeri). The presene of flvonois, tnnins, phenolis, sponins n other phytohemils in the queous root rk extrt of this plnt hs een reporte [6]. T. viennioies is known s Bushe (Hus, Northern Nigeri), i (Yoru) n Eo (go, Southest Nigeri). The roots ontin phenolis, tnnins, sponins n lklois [6]. Previously, stuies hve een rrie out on these plnts. Aqueous root rk extrts of A. leiorpus n T. viennioies s well s their mixture hve een reporte to inhiit the prolifertion of Ehrlih site rinom ells in vitro n to possess ntioxint properties [6, 7]. The plnts n their mixture hve een reporte to inrese the tivities of ntioxint enzymes n other non-enzyme ntioxints in the liver of norml rts s well s inhiit free rils in vitro [7]. Their protetive effets on DENmeite flututions of liver enzymes n funtionl inies hve lso een reporte [8]. Stuies on the effets of these plnts n their mixture on hepti reox sttus n hemtologil prmeters of DEN-trete rts hve not een reporte. Due to the ft tht mn is expose to vriety of ngerous hemils tht inue oxitive stress in the system, this stuy ws esigne to investigte the protetive effets of queous root rk extrts of A. leiorpus n T. viennioies s well s their mixture on hepti reox sttus n hemtologil prmeters of DEN-trete rts using urumin s referene ntioxint. MATERALS AND METHODS Plnt Mterils Fresh roots of A. leiorpus n T. viennioies were otine from frmln in Off, Nigeri in Deemer 2010 n uthentite t the FE Herrium, Ofemi Awolowo University, le-fe, Nigeri, with vouher numers n 15428, respetively. Experimentl Animls Mle lino rts (181.98±9.36 g) of Wistr strin were otine from the Animl House of Deprtment of Biohemistry, University of lorin, lorin, Nigeri. The nimls were house in luminium ges ple in well-ventilte house with optimum onitions (temperture: 22 Amu Kyoe Slu et l ±3 0C; photoperio: 12 hours nturl light n 12 hours rk; humiity: 40-45%) n were llowe free ess to rt pellets (Premier Fee Mill Co. Lt., n, Nigeri) n tp wter. The stuy ws onute t the Biohemistry n Nutrition Unit of Deprtment of Chemil Sienes, Fountin University, Osogo, Nigeri. Chemils n Regents itmin C, DEN, urumin n glutthione were prouts of Sigm-Alrih, St. Louis, USA. Assy kits for the enzymes superoxie ismutse (SOD), tlse (CAT), glutthione peroxise (GPx), glutthione reutse (GRe), n gluose 6-phosphte ehyrogense (G6PDH) were prouts of Rnox Lortories, UK. All other regents were of nlytil gre n prepre in ll glss-istille wter. Preprtion of Extrts The queous extrts were prepre s esrie previously [6-8]. Fresh root rks of oth plnts were seprtely oven-rie t 40 oc for 3 weeks n pulverise with n AKRA lener (moel: BL-1531, nonesi). A known mount (300 g) of eh power ws extrte in 5 L of istille wter n ple on oritl shker mintine t 300 rpm for 24 hours. This ws then filtere with Whtmn No. 1 filter pper n the filtrte onentrte on wter th to yiel 28.5 g of A. leiorpus (9.5%) n 30 g of T. viennioies (10%). A 1:1 (w/w) mixture of oth extrts ws lso prepre for use in the present stuy. Animl Grouping n Preprtion of Liver Superntnts Eighty-four mle lino rts were rnomly ssigne into twelve groups (A-L) of seven rts eh n trete orlly with 0.5 ml of istille wter, extrts or their mixture (1:1 w/w) ily for 28 ys s follows: Group A - reeive istille wter only (ily) for 4 weeks; Group B reeive istille wter (ily) for 4 weeks n DEN (30 mg/kg oy weight in norml sline) on y 15 n y 22 (weeks 3 n 4); Group C pre-trete with urumin referene ntioxint gent (200 mg/kg oy 22 ol 10, No 1, Jn-Fe 2016;

3 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 Effets of Aqueous Root Brk Extrts of weight in olive oil) ily for 4 weeks n DEN (s in group B ove); Group D pre-trete with 200 mg/kg oy weight A. leiorpus extrt ily for 4 weeks n DEN (s in group B ove); Group E - pre-trete with 400 mg/kg oy weight A. leiorpus extrt ily for 4 weeks n DEN (s in group B ove); Group F - pre-trete with 800 mg/kg oy weight A. leiorpus extrt ily for 4 weeks n DEN (s in group B ove); Group G - pre-trete with 200 mg/kg oy weight T. viennioies extrt ily for 4 weeks n DEN (s in group B ove); Group H - pre-trete with 400 mg/kg oy weight T. viennioies extrt ily for 4 weeks n DEN (s in group B ove); Group - pre-trete with 800 mg/kg oy weight T. viennioies extrt ily for 4 weeks n DEN (s in group B ove); Group J - pre-trete with 200 mg/kg oy weight of the mixture ily for 4 weeks n DEN (s in group B ove); Group K - pre-trete with 400 mg/kg oy weight of the mixture ily for 4 weeks n DEN (s in group B ove); n Group L - pre-trete with 800 mg/kg oy weight of the mixture ily for 4 weeks n DEN (s in group B ove). The rts were srifie 24 hours fter the lst oses y pling them in glss jr ontining otton wool soke in iethyl ether efore their jugulr veins were ut for loo olletion. Bloo ws ollete into len ottles ontining ntiogulnt for hemtologil nlysis. Liver superntnts were prepre s previously esrie [8, 9] n use for the iohemil ssys. The stuy ws pprove y the Ethil Committee on the Cre n Use of Experimentl Animls of the College of Nturl n Applie Sienes, Fountin University, Osogo, Nigeri. The nimls were lso hnle oring to the Guielines of the Ntionl nstitute of Helth (NH; Bethes, Mryln, USA), Guie for the Cre n Use of Lortory Animls [10]. Biohemil Assys n Hemtologil Anlysis The iohemil ssys were rrie out using stnr methos s esrie for rnin Journl of Toxiology mlonilehye (MDA) [11], SOD [12], tlse [13], GPx [14], GRe [15], G6PDH [16], glutthione [17] n vitmin C [18]. Hemtologil prmeters, nmely re loo ells (RBC), hemogloin onentrtion (H), pke ell volume (PC), men ell volume (MC), men ell hemogloin (MCH), men ell hemogloin onentrtion (MCHC), white loo ells (WBC), neutrophils (NEU), lymphoytes (LYM) n pltelets (PLT) were etermine on hemtology nlyzer, SysmexHemtology Systems (SysmexAmeri n., moel no. KX-21N, Koe, Jpn). Sttistil Anlysis The group men (n = 7) ±S.D for eh nlysis ws lulte n signifint ifferenes were etermine y Anlysis of rine (ANOA) n Tukey s Post Ho test for multiple omprisons t 95% onfiene level using SPSS softwre (SPSS n., Chigo, L, USA). RESULTS Liver MDA in DEN-trete rts ws signifintly (P<0.05) elevte ut pre-tretment with A. leiorpus t 800 mg/kg oy weight proue MDA levels tht ompre fvourly with istille wter- n urumin-trete ontrol nimls (Tle 1). n ition, ll the oses of the mixture of the extrts proue MDA levels tht ompre fvourly (P>0.05) with the urumin- n istille wter- trete nimls (Tle 1). DEN signifintly (P<0.05) reue the tivities of liver SOD, tlse, GPx, GRe n G6PDH of the nimls (Figures 1-5). This tren of reutions were signifintly (P<0.05) reverse following pre-tretment of the nimls with the extrts n their mixture (Figures 1-5) in ose-epenent mnner. nterestingly, the 800 mg/kg oy weight of the iniviul extrts n their mixture proue SOD n tlse tivities tht ompre well with the urumin- n istille wter-trete nimls (Figures 1 & 2) exept for the 200 mg/kg oy weight of A. leiorpus n the 200 n 400 mg/kg oy weight of T. viennioies (Figure 3). The 200 mg/kg oy weight of A. leiorpus proue GPx tivities tht were still signifintly (P<0.05) less thn the istille wter- n urumin-trete nimls (Figure 4) ol 10, No 1, Jn-Fe 2016

4 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 rnin Journl of Toxiology The 400 mg/kg oy weight of the mixture lso proue G6PDH tivity, whih ompre well with the ontrols (Figure 5). Reue glutthione (GSH) n vitmin C onentrtions were signifintly (P<0.05) erese following the ministrtion of DEN. These reutions were signifintly (P<0.05) reverse fter pre-tretment with the extrts n their mixture, though the vlues proue were still lower thn those of the ontrols (Tle 1). The elevtions in the levels of GSH y A. leiorpus n T. viennioies were not omprle with tht of urumin (Tle 1). Furthermore, the 800 mg/kg oy weight of the mixture of the extrts proue levels of GSH tht ompre well with tht of urumin (Tle 1). The 400 mg/kg oy weight of A. leiorpus n the 800 mg/kg oy weight of the iniviul extrts lso proue vitmin C onentrtions tht ompre well with the istille wtertrete ontrol nimls, while 400 n 800 mg/kg oy weight of mixture of the extrts proue vitmin C onentrtions tht were omprle to the urumin-trete nimls (Tle 1). Aministrtion of DEN n the extrts i not signifintly (P>0.05) lter the H, RBC, PC, MC, MCH, MCHC, WBC, LYM n NEU when ompre with the ontrol (Tle 2). PLT were, however, signifintly (P<0.05) elevte following the ministrtion of DEN n this ws reverse y the iniviul extrts n their mixture. The 800 mg/kg oy weight of ll the extrts s well s the 400 mg/kg oy weight of the mixture of the extrts proue pltelet ounts tht ompre fvorly with the urumin- n istille wtertrete nimls (Tle 2). Amu Kyoe Slu et l Tle 1.Liver mlonilehye (MDA), reue glutthione n vitmin C ontents of iethylnitrosmine (DEN)-trete rts pretrete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. A. leiorpus T. viennioies Mixture Liver mlonilehye (nmol MDA/mg protein) 3.05± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.18 Liver reue glutthione (nmol/mg protein) 30.00± ± ± ± ± ±0.45 e e 36.72± ± ± ± ± ± ± ± ± ± ± ±0.37 Liver vitmin C (mg/l) 54.09± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±1.61 lues re men of 7 etermintions±stnr Devition. lues rrying ifferent supersripts own the olumn for eh prmeter re signifintly ifferent (P<0.05). A. leiorpus = Anogeissus leiorpus, T. viennioies = Terminli viennioies, = ontrol, = Diethyl nitrosmine (30 mg/kg wt), = DEN+Curumin (200 mg/kg wt), = DEN+Extrt (200 mg/kg wt), = DEN+Extrt (400 mg/kg wt), = DEN+Extrt (800 mg/kg wt) Figure 1.Liver superoxie ismutse tivities of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Brs rrying ifferent letters for eh tretment re signifintly ifferent (P<0.05). 24 ol 10, No 1, Jn-Fe 2016;

5 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 Effets of Aqueous Root Brk Extrts of rnin Journl of Toxiology Figure 2.Liver tlse tivities of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Brs rrying ifferent letters for eh tretment re signifintly ifferent (P<0.05). Figure 3.Liver glutthione peroxise tivities of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Brs rrying ifferent letters for eh tretment re signifintly ifferent (P<0.05). Figure 4.Liver glutthione reutse tivities of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Brs rrying ifferent letters for eh tretment re signifintly ifferent (P<0.05). Figure 5.Liver gluose 6-phosphte ehyrogense tivities of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Brs rrying ifferent letters for eh tretment re signifintly ifferent (P<0.05) ol 10, No 1, Jn-Fe 2016

6 rnin Journl of Toxiology Amu Kyoe Slu et l Tle 2.Hemtologil prmeters of iethylnitrosmine (DEN)-trete rts pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture. Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 A. leiorpus T. viennioies Mixture A. leiorpus T. viennioies Mixture Hemogloin (g/l) Re Bloo Cells (x106/µl) 12.80± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.31 Pke Cell olume (%) Men Cell olume (fl) 45.23± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±1.05 Men Cell Hemogloin (pg) Men Cell Hemogloin Con. (g/l) 18.20± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± White Bloo Cells (x10 /µl) Lymphoytes (%) 15.87± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± Pltelets (x10 /µl) Neutrophils (%) ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.92 lues re men of 7etermintions±Stnr Devition. lues rrying ifferent supersripts own the olumn for eh prmeter re signifintly ifferent (P<0.05). A. leiorpus = Anogeissus leiorpus, T. viennioies = Terminli viennioies, = ontrol, = Diethyl nitrosmine (30 mg/kg wt), = DEN+Curumin (200 mg/kg wt), = DEN+Extrt (200 mg/kg wt), = DEN+Extrt (400 mg/kg wt), = DEN+Extrt (800 mg/kg wt). DSCUSSON Rts were pre-trete with queous root rk extrts of A. leiorpus, T. viennioies n their mixture (1:1 w/w) n were therefter hllenge with DEN, potent heptotoxin, whih inue ltertions in reox prmeters of the liver n some hemtologil inies. The extrts prevente the DEN-inue effets in mnner omprle with urumin, referene ntioxint. MDA is eomposition prout of lipi peroxies, whih result from lipi peroxition [19]. t is n initor of lipi peroxition n hs een use s n inex of tissue toxiity [20]. MDA in the ell n ttk intrellulr mromoleules, inluing DNA, leing to mutgeniity n rinogeniity [4]. The signifint inrese in liver MDA onentrtion y DEN might inite inrese peroxition of the unsturte ftty is in the ellulr memrne of the liver. DEN proues exessive ROS [5], whih ttk the unsturte ftty is in the ellulr memrne therey using lipi peroxition. The ility of the extrts n their mixture to ttenute the lipi peroxition is n inition tht they oul hve loke the lipi peroxition pthwy y ting s ntioxints. 26 ol 10, No 1, Jn-Fe 2016;

7 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 Effets of Aqueous Root Brk Extrts of This might e ttriute to the presene of tnnins, flvonois n phenolis, whih re reportely present in the extrts [6] n might hve te s ROS svengers n hin reker in the proess of lipi peroxition. Reution of ellulr MDA woul lso reue the risk of ner. nrese in MDA onentrtion y DEN reporte in this stuy is similr to previous finings [2, 4]. DEN hs een reporte to inue oxitive stress n retrogrtion of the ntioxint efene mehnism ue to overproution of ROS, lipi peroxition n mge to mny enzymes involve in DNA repir, whih result in orgn isorers, espeilly the liver [2, 5]. nrese ROS proution y DEN woul shift the reox lne of the ell in fvour of ROS, therey epleting the ntioxint system. The reox sttus of ells is mintine y set of enzymi n non-enzymi ntioxints. The ntioxint enzymes SOD, CAT, GPx, GRe n G6PD re involve in oorinte proess to prevent oxitive mge y ROS. SOD ismuttes superoxie ril to H2O2, whih is further eompose to wter n oxygen y tlse [19]. GPx tlyzes the reution of hyroperoxie (ROOH) with onomitnt oxition of glutthione. The oxiize glutthione is then reue y GRe, oxiizing NADPH in the proess. G6PDH onverts the oxiize NADP+ to the reue form (NADPH) y oxiizing gluose 6-phoshte. This oorinte set of retions eventully le to reution of ellulr ROS onentrtion. The exessive proution of highly oxiize n oxiizing free rils in ells hs een vriously reporte to reue the tivities of these enzymes. Furthermore, the inution of these enzymes y hemil ompouns hs lso een reporte to e ssoite with in vivo ntioxint tivity [4, 7]. The reution in the tivities of SOD y DEN is n inition of exessive ROS proution. Sine SOD onverts superoxie ril to H2O2, reue SOD tivity will le to the prtiiption of superoxie in the Fenton Retion to proue hyroxyl rils, whih will oxitively mge DNA, lipis n proteins [19] n my le to the evelopment of ner. The ility of the extrts to reverse this tren is n inition of ytoprotetive tivity ginst ROS n ntioxint tivity of the extrts s reporte in previous stuies [6, 7]. Ctlse tlyzes the eomposition of H2O2 to wter. The reution in the tivity of rnin Journl of Toxiology this enzyme y DEN woul lso le to the prtiiption of hyrogen peroxie in the Fenton Retion to proue hyroxyl rils s mentione erlier in this stuy. The ility of the extrts to reverse this DEN-inue reution in tlse tivity is n inition of in vivo ntioxint tivity ginst oxitive stress. GPx tlyses the etoxifition of orgni hyroperoxies n H2O2. t lso tlyses the reution of isulphie ons in proteins y glutthione. Glutthione is onsequently oxiise to its oxiise form (GSSG). DENinue reution of GPx tivity will le to the umultion of H2O2 n orgni hyroperoxies in the ells. This my onsequently le to oxitive stress. The ttenution of this tren y the extrts further inites ntioxint tivity. GRe tlyses the onversion of GSSG to GSH using NADPH n, in the proess, provies protetion to ellulr onstituents ginst ROS. The reution of the tivity of this enzyme y DEN will le to ROS-inue oxitive mge. The ility of the extrts to reverse this tren is lso n inition of ntioxint tivity. G6PDH reyles the NADPH onsume y GRe so tht ellulr GSH pool n e replenishe. DEN-inue erese in G6PDH tivity will le to umultion of NADP+ n GSSG s well s epletion of GSH, whih n le to oxitive stress. The reversl of this DEN-inue oxitive stress y the extrts is lso n inition of in vivo ntioxint tivity. The non-enzyme ntioxints glutthione n vitmin C t in omplementry mnner to the ntioxint enzymes. Glutthione reues the isulphie ons of ytoplsmi proteins to ysteines, therey forming GSSG, whih is reue y GRe to GSH. Cellulr glutthione onsists of GSSG n GSH; inrese in GSSG/GSH rtio is n inition of oxitive stress. The erese in the levels of reue glutthione following DEN ministrtion is suggestive of oxitive stress. itmin C or sori i ontes eletrons to oxints, forming the stle ehyrosorte, n therefore, ting s n ntioxint. The DEN-inue erese in the vitmin C ontent will mke the ells prone to ttk y ROS. The reversl of this tren y the extrts further orroortes the erlier reporte ntioxint tivity of the extrts. DEN-inue erese in the enzymi n non-enzymi ntioxint systems woul le to 27 ol 10, No 1, Jn-Fe 2016

8 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 rnin Journl of Toxiology mge of ellulr iomoleules, inluing DNA, y ROS, whih oul le to mutgenesis n then rinogenesis. The ttenution of this DEN-inue mge y the extrts oul men tht the extrts my t s ner preventing gents vi ntioxint mehnism. This further orroortes the reution in MDA, reporte in this stuy. These results gree with previous reports [2, 4, 7] tht DEN uses erese in ntioxint systems, n tht extrts possessing in vivo ntioxint n ntirinogeni tivities n reverse suh proesses n inue the inrese in onentrtions of ellulr ntioxints. These tivities oul e ttriute to the ntioxint phytohemils previously reporte to e present in the extrts [6]. The preventive tivities exhiite y the extrts were, in most of the ses, proue y the 800 mg/kg oy weight of the iniviul extrts, whih ompre well with the referene ntioxint urumin n the ontrol. The mixture, espeilly, proue the omplete reversl of the DEN-inue liver mge.hemtologil prmeters n e use to monitor intertions of hemils with the iologil system in vivo in orer to ssess their systemi effets. RBCs ontin hemogloin, whih rries oxygen to the tissues n returns CO2 to the lungs uring respirtion. RBC, H n MCH re ll inies of re loo ells n their reution n inite nemi just s n inrese n inite inrese rte of erythropoiesis, i.e. re loo ell synthesis [21]. White loo ells efen the oy ginst infetions or ny foreign oy. Elevte vlues my inite oost in immunologil tivity or pthologil onition [22]. The lk of ny effet on these prmeters y DEN is n inition tht the lne etween the rtes of synthesis n estrution of these loo ells were not ffete. t lso mens tht the immune system of the nimls ws not ffete. These finings re in ontrst with tht of previous stuy [23], whih reporte reution in RBC, H, PC, MC, MCH, MCHC n totl white loo ell ount y single intrperitonel ministrtion of 200 mg/kg oy weight of DEN. This ifferene might e relte to ifferenes in oses n routes of ministrtion of DEN. Pltelets tivte the loo lotting mehnism [24] n funtion minly in the formtion of mehnil plugs uring norml hemostti response to vsulr injury [25]. The elevte pltelet ounts oul hve resulte Amu Kyoe Slu et l from liver injury use y DEN. The reversl of this elevtion of pltelet ount y the extrts further inites tht the extrts possess hemoprotetive effets. This oul e ue to the presene of flvonois in the extrts, whih hve een implite in the prevention of pltelet ggregtion [26]. This stuy is limite y the ft tht the tive ingreients hve not yet een ientifie, whih woul provie iretion for the isoltion n proution of potent heptoprotetive gents in the future. CONCLUSON Aqueous root rk extrts of A. leiorpus n T. viennioies s well s their mixture possess ntioxint tivities exhiite y their heptoprotetive n hemoprotetive tivities ginst hepti reox n hemostti imlnes use y DEN in rts. These tivities hve een ompre well with urumin n etter exhiite y the mixture thn the iniviul extrts. ACKNOWLEDGEMENT This pper is n spet of Dotorl (Ph.D.) thesis title: Antiner Ativity n Toxiologil Effets of Aqueous Extrts of Anogeissus leiorpus n Terminli viennioies Root Brks, Deprtment of Biohemistry, University of lorin, lorin, Nigeri (Slu 2013; Unpulishe). This stuy reeive no grnts n the uthors re the only finnil soure for the reserh. The uthors re grteful to the University Mngement n lortory stff of Deprtment of Chemil Sienes, Fountin University, Osogo, Nigeri. The uthors elre tht there re no onflits of interests. REFERENCES 1. Ym K, Ymmiy, Utsumi H. n vivo etetion of free rils inue y iethylnitrosmine in rt liver tissue. Free Ril Biol Me 2006;40: Chen B, Ning M, Yng G. Effet of peonol on ntioxint n immune regultory tivity in heptoellulr rinom rts. Moleules 2012;17: Sheweit SA, Mous N, Newiry AA. Nnitrosoimethylmine inue hnges in the tivities of rinogen-metolizing enzymes in the liver of mle mie: role of glutthione n gossypol s ntioxints. Afr J Biohem Res 2007;1(5): ol 10, No 1, Jn-Fe 2016;

9 Downloe from ijt.rkmu..ir t 13: on Stury Jnury 12th 2019 Effets of Aqueous Root Brk Extrts of 4. Pl R, Thirumoorthy N. Chemo-preventive effet of ethnoli extrt of Morin itrifoli root on N-nitrosoiethylmine inue oxitive stress in rt liver. Worl J Phrmeutil Res 2012;1(3): Al-Rejie SS, Aleis AM, Al-Yhy AA, Bkheet SA, Alsheikh A, Ftni AG, et l. Progression of iethylnitrosmine-inue hepti rinogenesis in rnitine eplete rts. Worl J Gstroenterol 2009;15(11): Slu AK, Ykuu MT, Oliji AT. Cytotoxi tivity of queous extrts of Anogeissus leiorpus n Terminli viennioies root rks ginst Ehrlih Asites Crinom ells. nin J Phrmol 2013;45: Slu AK, Ykuu MT, Oliji AT. n vitro n in vivo ntioxint tivity of queous extrts of Anogeissus leiorpus (DC) Guill & Perr n Terminli viennioies Guill & Perr root rks. Cmeroon J Biol Biohem Si 2015;23: Slu AK, Ykuu MT, Oliji AT. Heptoprotetive Effets of Aqueous Root Brk Extrts of Anogeissus leiorpus, Terminli viennioies n their Mixture in Diethylnitrosmine-Trete Rts. Drug Dev Therpeutis 2015;6(2): Ykuu MT, Aeshin AO, rhim OOK. Toxiologil evlution of queous lef extrt of Senn lt in pregnnt Wistr rts. ntl J Curr Res Rev 2012;4(8): Ntionl nstitutes of Helth. Guie for the re n use of lortory nimls. NH Pulition 1985;83: Buege JA,Aust SD.Mirosoml lipi peroxition.methos Enzymol 1978;52: Woollims JA, Wiener G, Anerson PH, MMurry CH. Res eter Si 1983;34: Beers RF Jr, Sizer W.A spetrophotmetri metho for mesuring rekown of hyrogen peroxie y tlse. J Biol Chem 1952;195: Rotruk JT, Pope AL, Gnther HF, Swson AB. Selenium: iohemil role s omponent of glutthione peroxise. Siene 1973;179: rnin Journl of Toxiology 15. Golerg DM, Spooner RJ. Methos of Enzyme Anlysis n: Bergmeyen H, eitor, 1983; 3r e. p Lohr GW, Wller HD. Gluose 6-phosphte ehyrogense. n: Bergmeyer HU, eitor, Methos of Enzymti Anlysis. New York: Aemi Press; p Ellmn GL. Tissue sulfhyryl groups. Arh Biohem Biophysis 1959;82: Omye ST, Turull TP, Suerhih HC. Selete methos for etermintion of sori i in ells, tissues, n fluis. Methos Enzymol 1979;6: Liou G-Y, Storz P. Retive oxygen speies in ner. Free Ril Res 2010;4(5): Krulut-Buln O, Bolkent S, Ynrg R, Bilgin-Sokmen B. The role of vitmin C, vitmin E, n selenium on mium-inue renl toxiity of rts. Drug Chem Toxiol 2008;31: Gnong WF. Review of Meil Physiology. 20th e. Lonon: Lnge Meil Books/Mgrw Hill Meil Pulishing Division; p Qio ZS, Wu H, Su ZW. Comprison with the phrmologil tions of Morin offiinlis, Dmnnthus offiinrium n Shisnr propinqu whose roots hve een use sine nient time. Zhong Xi, Yi, Jie he, Z Zhi 1991;11(7): Bishyee A, Krmkr R, Mnl A, Kunu SN, Chtterjee M. nium-meite hemoprevention ginst hemil heptorinogenesis in rts: hemtologil n histologil hrteristis. Europ J Cner Preven 1997;6(1): Psternk CA. An ntroution to Humn Biohemistry. USA: Churhill Livingstone; Hoffrn A, Pettit JE, Moss PAH. Essentil Hemtology, 4th e. itori: Blkwell Siene Asi Ply Lt; Okwu DE, Okwu ME. Chemil omposition of Sponis momin Linn plnt prt. J Sust Agri Env 2004;6(2): ol 10, No 1, Jn-Fe 2016