Protective Efficacy of Spirulina platensis Against Cadmium Induced Neurotoxicity in Rats

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1 Glol Veterinri 14 (4): , 2015 ISSN IDOSI Pulitions, 2015 DOI: /iosi.gv Protetive Effiy of Spirulin pltensis Aginst Cmium Inue Neurotoxiity in Rts 1 2 Rsh T.M. Alm n M.Y. Henwi 1 Deprtment of Clinil Pthology, Fulty of Veterinry Meiine, Zgzig University, Egypt 2 Deprtment of Plnt Protetion, Fulty of Agriulture, Zgzig University, Egypt Astrt: Cmium (C) is potent neurotoxi hevy metl, whih inues oxitive stress n memrne isturnes in rin. The present stuy investigte the possile neuroprotetive effiy of Spirulin pltensis (Sp) ginst CCl 2 inue hnges in the plsm neurohemistry of teholmine (renline, nor renline, opmine), serotonin, gmm mino-utyri i (GABA), tivity of etyl holinesterse (AChE), DNA frgmenttion of the rin ells n the histopthology of the rin tissue in rts. Animls were ivie into five groups. Gp.(1) Control, gp.(2) CCl 2 (2mg/kg wt) S/C injetion, gp.(3) Spirulin (150 mg/kg wt) orlly, gp(4) Co-trete with oth CCl 2 n Sp n gp.(5) Aministere CCl 2(2mg/kg wt), in ll gps the tretment is y fter y for 30 ys exept gp.(5) post- trete y Sp for nother 30 ys. The results inite tht mium inue oxitive stress in the rin leing to ltertions in the plsm neurohemistry n inrese the DNA frgmenttion of the rin tissue, Spirulin pltensis given inomintion n post-tretment with C signifintly reues pthologil ltertions in plsm neurohemistry, erese the egree of DNA frgmenttion of the rin ells n ple of reuing the pthologil mge of the rin tissue. We onlue tht S.pltensis n signifintly moify the rin mges ginst mium hlorie inue toxiity. Key wors: Cmium, Spirulin Neurotoxiity DNA Frgmenttion Cteholmine GABA INTRODUCTION (0.5 n 1mg/kgm wt) erese opmine level in the ifferent rin res [7]. Cmium inreses the serotonin Cmium (C) is toxi metl whih still ttrts the sensiility in the CNS [8]. Assoite hnges in ttention of reserhers n the puli helth euse its norepinephrine, opmine n serotonin ontent n/or level in the foo prouts my e exees the mximum its metolism in the ifferent rin regions were llowle limits. The hrmful effets of mium in mny oserve in mium expose rts [9], [10,11]. While orgns n its exposure ssoite with nephrotoxiity, GABA ontent in ifferent rin regions erese in osteoporosis, neurotoxiity, rinogeniity, genotoxiity, rts given 50 ppm of mium hlorie for 1 month [12]. tertogeniity, enorine n reproutive sie effets Some stuies on C toxiity foun n ssoition with [1, 2]. Exposure to C severely ffets the funtion of the ehviorl isturnes n holinergi nervous system [3,4]. neurotrnsmission sine n inrese or erese in the Moreover, exposure to mium (0.25&1 mg/kg/y) etylholinesterse tivity ws verifie in oth niml for 45 ys moifies the CNS regionl ontent of moels n humn tht showe ehviorl impirments etylholine (ACh), serotonin (5-HT) n opmine [5]. fter exposure to C [13]. AChE tivity in the rin Rts ily s/ injete with CCl 2 (0.3 mg/100g wt) for inrese in rts ministere mium (1 mg/kg/y for two weeks in norml n protein restrite (5% sein) 14 ys/ ip) [14]. Rts expose to C (3mg/kg/y s) for groups shows signifint erese in 5-HT onentrtion 3 weeks, signifint inrese in the tivities of AChE in ifferent rin res [6]. Also mium expose rts were oserve in rin tissue [13]. Cmium intoxite Corresponing Author: Mohme Yossif Irhim Henwi, Deprtment of plnt protetion, Fulty of Agriulture, Zgzig University, Egypt. E-mil: Henwi.m@zu.eu.eg. 490

2 rts (5mg/kg/y y gvge) for 4 weeks signifintly reue the AChE levels in the plsm n rin tissue [15]. Also the AChE tivity inrese in the hippompus, hypothlmus n ereellum of hroni C-intoxite rts (25mg/kg) [16]. Spirulin pltensis (SP) is ynoterium eing use s nutritionl supplement for humn n niml onsumption. Spirulin pltensis hs een lele s powerful foo, rih in proteins, rohyrtes, polyunsturte ftty is, sterols n some vitl elements like lium, iron, zin, mgnesium, mngnese n selenium. It is nturl soure of vitmin B12, vitmin E, sori i n rotene n xnthophyll phytopigments [17,18]. Spirulin hs een evlute for vrious tivities inluing neuroprotetion in roents [19,20]. In ition, severl stuies hve shown tht Spirulin speies exhiit vrious iologil tivities suh s ntioxint, ntiteril n ntiprsiti properties n for severl meil onitions suh s llergies, ulers, nemi, hevy-metl poisoning n rition poisoning [21,22]. Moreover, omine tretment of Spirulin with meruri hlorie le to signifint erese in MDA ontent n elevtion in LDH n ALP tivity, it moify the renl mges ginst meruri hlorie inue toxiity [23]. Also tretment with Spirulin pltensis elevte the levels of FT3, FT4 n TSH, Spirulin filitte ntioxint formtion, improve ehviorl hnges n thyroi ysfuntion in fluorie toxiity in rts [24]. Sine SP hs mny therpeuti roles inluing neuroprotetive properties, the present stuy ime to ientify the protetion exerte y Spirulin pltensis ginst mium inue neurotoxiity in the rts. MATERIALS AND METHODS Animls: A totl of 25 helthy ult mle white lino rts ( gm oy weight). They were otine from the Lortory Animl Housing Unit, Reserh, Institute, Dokki, Ciro, Egypt. Animls kept uner hygieni onitions. They were mintine on sl rtion, given wter -liitum n expose to 12h light-rkness yle for two weeks of limtiztion efore use. Teste Compouns Cmium Chlorie (CCl 2): Purhse from El-Frn Co. for tring, Egypt in the form of white n olorless power. Spirulin Pltensis (Sp) Power: Ws purhse from El Hellow for Biologil prouts Egypt. Experimentl Design: The rts were ssigne into five equl groups eh ontining 5 rts. Gp.(1) kept s ontrol, gp.(2) ws given CCl 2 (2mg/kg wt) S/C injetion, gp.(3) ministere Spirulin power (150 mg/kg wt) in rinking wter, gp.(4) rts o-trete with oth C n Sp t the sme mentione routes n oses y fter y for 30 ys in the mentione groups n gp.(5) ministere CCl 2 (2mg/kg wt) y fter y for 30 ys with post- tretment y Sp (150 mg/kg wt) for nother 30 ys. Smples Colletion n Preprtion: At the en of the experiment the nimls were srifie, followe y olletion of loo without ntiogulnt then entrifuge t 3000 rpm for 10 min for seprtion of serum whih store t -20 C until use for iohemil nlysis. Brins were issete out n wshe in sline. Brin tissues were homogenize in oule istille wter n the homogentes were store t -80 C until use. Other smples from rin were use for histopthologil exmintion. Biohemil Anlysis: Dopmine, renline, norrenline, Serotonin, Aetylholinesterse tivity (myiosoure.o) n GABA (EIA.o) were mesure using n ELISA kit. A iphenylmine olorimetri ssy (Rouh.o) ws use to quntify DNA frgmenttion, the prepre rin homogente were entrifuge t 300 g for 10 min to seprte superntnt from ell pellets. One hunre µl liquots of superntnts were use for DNA frgmenttion ssy. Histopthologil Investigtion: Speimens from the rin of ifferent groups were ollete n fixe in 10% uffere neutrl formlin solution, ehyrte in grul ethnol (70-100%), lere in xylene n emee in prffin. Five-miron thik prffin setions were prepre n then routinely stine with hemtoxylin n eosin (H&E) yes [25] n then exmine mirosopilly. Sttistil Anlysis: Dt of the urrent stuy expresse s men ± stnr error ws sttistilly nlyze using the omputer progrm SPSS/PC (2001). The sttistil metho ws one wy ANOVA test. 491

3 RESULTS Figure (1) represents the quntittive mesurement DNA frgmenttion. In mium trete group, Tle (1) epits the effets of C n SP on the initing signifint (p<0.05) inrese in the DNA plsm teholmine (renline, nor renline n frgmenttion, while ministrtion of Sp lone showe opmine) levels, serotonin, GABA, AChE tivity in non signifint hnges when ompre with ontrol rts. ontrol n experimentl rts. In C trete rts (gp.2), the Tretment with Sp (gps. 4 n 5) showe signifint teholmine (renline, nor renline n opmine) (p<0.05) erese in the DNA frgmenttion when levels signifintly (p<0.05) inrese when ompre with ompre with gp.(2). ontrol rts. Aministrtion of Spirulin lone (gp.3) proue non signifint hnges in the teholmine, Histologil Chnges in Brin: Figures (2, 3,4,5,6 &7) while tretment with Sp (gps.4 n 5) effetively illustrte the histopthologil ssessment of rin ttenute the C-inue ltertions in levels of tissue of ontrol n experimentl nimls. C teholmine. A signifintly (p<0.05) erese in the intoxite rts exhiite ererl hemorrhge (Fig. 3), plsm serotonin n GABA levels in gp.(2), while gp.(3) epletion of grnulr ell lyer (rrowhe) n showe non signifint hnges when ompre with neroti hnges in the Purkinje ells (Fig. 4) n ilte ontrol. Tretment with Sp (gps.4 n 5) showe lterl ventrile, suventriulr spongiosis n gliosis signifint inrese in the levels of serotonin n GABA (Fig. 5). Aministrtion of Sp lone the rin tissue when ompre with gp.(2). The tivities of AChE in the showing norml neurons n neuronoglis (Fig. 2). plsm ws signifintly (p<0.05) inrese in gp.(2) n Tretment with Sp (gps.4 n 5) the rin tissue showing gp.(3) showe non signifint hnges when ompre few egenerte neurons, stellitosis n with ontrol rts, while gps.(4&5) signifintly (p<0.05) neuronophgi (Fig. 6) n moerte egenerte erese the tivities of AChE to ner norml levels neurons, stellitosis n neuronophgi (Fig. 7) when ompre with gp.(2). respetively. Tle 1: Effet of orl ministrtion of Spirulin pltensis (150mg/kg) on the plsm levels of some neurologil prmeters in C trete rts (2mg/kg) (M ±SE) (n=5) Groups ADPg/ml NADPg/ml Dopmineng/ml Serotonineng/ml GABAng/ml AChEu/L Control (Gp1) 18.10± ± ± ± ± ±1.23 C (Gp2) 28.94± ± ± ± ± ±1.33 Spirulin (Gp3) 18.59± ± ± ± ± ±0.56 Trete A (Gp4) 23.25± ± ± ± ± ±0.20 Trete B (Gp5) 26.03± ± ± ± ± ±0.27 Vlues re men ± SD for five rts in eh group. Vlues not shring ommon supersript letters (, n ) iffer signifintly t p<0.05 Fig. 1: Effet of orl ministrtion of Spirulin pltensis on DNA frgmenttion in rin tissue of C trete rts 492

4 Fig. 2: Brin setion of gp. (3) showing norml neurons n neuronoglis Fig. 3: Brin setion of gp. (2) showing ererl hemorrhge (rrow) Fig. 4: Brin setion of gp. (2) showing epletion of grnulr ell lyer (rrowhe) n neroti hnges in the Purkinje ells (rrow) Fig. 5: Brin setion of gp.(2) showing ilte lterl ventrile, suventriulr spongiosis (rrow) n gliosis (rrowhe) Fig. 6: Brin setion of gp.(4) showing few egenerte neurons, stellitosis n neuronophgi (rrows) Fig. 7: Brin setion of gp.(5) showing moerte egenerte neurons, stellitosis n neuronophgi (rrows) DISCUSSION neurohemistry ue to oxitive mge to the rin tissue whih my e responsile for its toxi tion Cmium (C) is n importnt environmentl [29,30]. Also C inue lipi peroxition y stimulting ontminnt ue to its extensive use in phosphte the proution of superoxie nions [31] whih t s fertilizers, eletrohemil inustry, irrft eletroplting, oxitive stress, this proess n mge ells iretly y nuler fusion, lortory uses n in everyy life estroying their memrnes n iniretly y prouing prouts [26], C is le to inue neurotoxiity inluing retive ronyl prouts [32]. The higher lipi neurologil isturnes with hnges in the norml peroxition inue y C inites the memrne neurohemistry of the rin [27]. Cmium exert toxi mge resulting in neuronl ysfuntion leing to effets on the entrl nervous system (CNS) [28]. erese the uptke of teholmine in rin In the present stuy, the plsm teholmine synptosomes [33] n inrese it in the plsm. Our (renline, nor renline n opmine) levels results re prtilly greement with [5] who reporte tht inrese in rts trete with mium (gp.2), it hs een hroni tretment with low oses of mium signifintly suggeste tht mium proue ltertion in the plsm inrese the level of stritl opmine in rts; lso [34] 493

5 foun tht ietes enhnes the effet of mium iomrker for the ssessment of C-inue neurotoxiity elevting the levels of opmine. Disrepnies with other [16]. This enzyme hyrolyses the neurotrnsmitter uthors [10,33] oul e ue to ifferenes in ge of etylholine (ACh) in the synpti left of holinergi nimls uring mium exposure, ose, route n synpses n neuromusulr juntions [46]. Cmium tretment urtion. signifintly inrese the AChE tivity, whih my e On the other hn, serotonin (5-HT) is hiefly present ttriute to the mehnism of tion of C on AChE in the hypothlmus, mirin n limi system of the enzyme whih hs een hypothesize to e either the rin [35,36], long with prllel istriution of isplement of metl oftors from the tive site or the tryptophn hyroxylse [37]. The plsm serotonin levels iret etivtion of the enzyme site [47]. Moreover, C signifintly erese in gp.(2) whih my e ttriute n les to ltertion of the struturl integrity of lipis to the non uniform umultion of C in the rin leing n ffets memrne-oun enzymes suh s AChE, to inhiition of numer of sulfhyryl (-SH) groups thus, ltertions in the lipi memrne y oxitive stress ontining enzymes [38], tht ffet the entrl oul e eisive ftor in the moifition of the AChE monominergi neurotrnsmitter system [39], Sine the moleule, whih woul explin the hnges in their permeility of the loo rin rrier of the mein tivity [48,49]. However, results re ontroversil n the eminene region lters rpily, it llows more C to enter, tivtion s well s the inhiition of the AChE tivity whih gretly ereses hypothlmi 5-HT level [40]. hs een reporte. Our results re prtilly greement with Also the mium tretment signifintly ereses the Crgeorgiou et l.,[ 50] who reporte tht the tivity of onentrtion of 5-HT in most of the isrete rin res, AChE in rt rin synptosoml plsm memrnes ue to interferes with the funtion of tryptophn showe onsierle erese fter 6 h of C exposure, hyroxylse whih plys n importnt role uring followe y progressive inrese up to 24 h. However, onversion of tryptophn to 5-HT utilizing oxygen y n inrese in the AChE tivity ws foun in rin of rts inhiiting the oxitive metolism of the serotonergi expose to 1 mg/kg C i.p for 14 ys or i.m for 4 months neurons [41]. Our results re prtilly greement with [15, 51]. While our results re isgreement with [52] Ds et l. [6] who reporte tht rts trete with CCl2 who reporte tht erese AChE tivity ws foun in shows signifint erese in 5-HT onentrtion in hippompus, ereellum n hypothlmus of rts ifferent rin res whih le to erese the plsm intoxite with 2 mg/kg C y gvge, the ifferene my level of serotonin. The present results re isgreement e ue to the hnge in the route of ministrtion. with Her et l. [8] who reporte tht mium inrese Shgirth et l., [15] lso reporte erese the level of the serotonin level in the rin, the ifferene my e ue AChE in plsm n rin in C expose rts (5mg/kg to hnge in ose or urtion n ehvior response of /orlly), the ifferene my e ue to lrge ose of C or the niml moels. ifferent route of ministrtion leing to ifferent GABA is onsiere s the min inhiitory response of the rts AChE tivity. neurotrnsmitter in the entrl nervous system [42]. Reent stuies hve shown tht mium proues GABA hs een ssoite with the evelopment of ROS, resulting in n inrese lipi peroxition, neuroegenertive iseses [43]. This results show tht epletion of sulphyryls, ltere lium homeostsis, rts trete with mium signifintly erese the impirment of ntioxint efenses n finlly DNA GABA level in the plsm whih my e ttriute to mge [53]. Uner the present experimentl onitions, erese the GABA ontent in the ifferent rin res it hs een oserve tht mium toxiity signifintly (the nterior n meiosl hypothlmus n mein inrese the DNA frgmenttion of the rin tissue, whih eminene [12]. Our t prtilly gree with Lfuente et l. my ttriute to the ility of umulte C in the [44] who reporte reution in the GABA ontent in rin to mge the rin ells ue to inrese lipi posterior hypothlmus n prefrontl ortex in rts peroxition [55], lso mium toxiity inrese the expose to mium. Also Soreq n Seimn [45] extent of DNA frgmenttion whih my e regre s reporte tht rts expose to C signifintly erese the n initor of inrese ROS proution uring toxin GABA ontent in the nterior n meiosl exposure perio [16]. The previous results re onfirme hypothlmus. with histopthologil exmintion of the rin tissue of The stuy of rin enzyme tivities, suh s AChE mium-trete rts (gp.2). Cmium inue is key enzyme in eteting the neurotoxi effet of neurotoxiity whih hrterize y ererl ertin hevy metls. AChE tivity is often onsiere hemorrhges sttere on the rin tissue (Fig. 3). 494

6 Depletion of the grnulr ells in the ereellum, esies whih ontins mny ntioxints ting on the oxitive some neroti hnges in the Purkinje ells (Fig. 4). stress. -rotene of Spirulin my svenge free rils Enephlomli, egenerte neurons, stellitosis n generte y C n reues the lipi peroxition. neuronophgi, sever perivsulr n perineurl eem The ntioxint mehnism of -rotene hs een esies ilte ventriles were visulize in ition to suggeste to e singlet oxygen quenhing, free ril suventriulr spongiosis n intense gliosis (Fig. 5). svenging n hin reking uring lipi peroxition Our results re greement with Mrles et l. [56] who [58,59]. reporte nuler onenstion of hromtin, shrunken Tretment with SP (gps.4 n 5) signifintly Purkinje ells n interstitil eem in moleulr lyer in improve the plsm levels of serotonin n GABA n rts trete with mium. Also Gonlves et l. [16] who restore the tivity of AChE when ompre with gp.(2). reporte tht the rin tissue revels tht mium An importnt spet of Spirulen s role in intoxition use norml ultr struturl hnges in neuroprotetion is ue to the inhiition of ril the rin tissue inluing spongiform nerosis, nuler formtion through its nti-inflmmtory effets. Selenium vuoliztion pyknosis n lymphoyti inflmmtory present in Spirulin-inue selenium ontining enzyme hnges. GSH peroxise, proteins or ompouns suh s In the present stuy, Spirulin Pltensis lone i selenoiglutthione, selenoysteine n not use ny sie effets or rin toxiity. Rts trete imethylselenie, whih re known to moulte the toxi with SP (gp.3) showe non signifint hnges in the effets of hevy metls [23]. The ynoteri Sp is n plsm neurohemistry (renline, norrenlin, exellent soure of phyoynin [60] ple of exerting opmine, serotonin, GABA n AChE tivity), it lso strong ntioxint tion, whih my e the use of protet the norml ells from mge y eresing the reuing the lipi peroxition n reues the egree of DNA frgmenttion in the rin ells when neurotoxiity in rts y effiiently interts with vrious ompre with the ontrol. The protetive n improving retive oxygen n nitrogen speies. Spirulin my e effets of Spirulin Pltensis on rin tissue my e ue ounterte the effets of C on the rin tissue y its to it hs vrious omponents, suh s B-omplex powerful ntioxint nture n y filitting the vitmins, hlorophyll, rotene, vitmin E, superoxie isplement of C, it reuing their umultion in the ismutse n numerous minerls, whih involve in oy. The ility of Spirulin to prevent peroxition of tissue protetion when ingeste [57]. Our results re lipis oul e linke to the presene of ntioxints onfirme y the histopthologil exmintion of the ( rotene n vitmin C, E). Chlorophyll n its rin tissue whih revele no signifint ifferene hs erivtives svenge free rils [61], so the presene of een oserve etween ontrol n Spirulin trete hlorophyll in Spirulin n ontriute to the ntioxint groups, (gp.3) whih showe nerly norml tion. Moreover Spirulin pltensis ontins -lipoi historhiteture (norml neurons n neuronoglis, i, rioflvin, xnthophyll phytopigments, SOD enzyme, Fig. 2. Spirulin Pltensis ws remrkly effetive in selenium, mgnesium n mngnese, whih n reuing the iniene of neurotoxiity n DNA ontriute to the ntioxint effet [62, 63]. frgmenttion in the rin use y oxitive stress Spirulin pltensis in omintion n post inue y C whih isrupts the integrity of the tretment in C toxite rts (gps.4&5) erese the neuronl ell memrne leing to ltertion in the plsm egree of the DNA frgmenttion when ompre with neurohemistry suggesting its potentil therpeuti effet gp.(2) my e ue to eresing the ell mge n in our moel. Our reserh tem hs lrey reporte tht proteting the ells y its powerful ntioxint tivity. rts trete with C in omintion n post trete with The present results re greement with [64] who reporte Spirulin Pltensis (gps.4 n 5) showe signifint tht rotene of Spirulin my reue ell mge, erese in the plsm teholmine (renline, espeilly the DNA moleules. Suh n inhiitory effet norrenlin n opmine) when ompre with C my e ttriute to the repir of rinogen- mge trete group (gp.2). This my e ttriute to the SP DNA n SP hs een suggeste s n effiient ril prevente mium from rehing suffiient svenger [65,66]. Other stuies hve reporte tht the onentrtion in the rin, so erese its mge n unique polyshries of SP enhne ell nuleus erese its toxi effet. The mehnism of this protetion enzyme tivity n potentite the proess of DNA repir seems to e se on the omplex moleulr struture [67, 68]. 495

7 The previous results of the trete groups with 4. Co, Y., A. Chen n J. Rliffe, Postntl spirulin pltensis were onfirme with histopthologil mium exposure, neuroevelopment n loo exmintion of the rin ells whih showe tht the pressure in hilren t 2, 5 n 7 yers of ge. lesions use y C in the rin in gp.(2) were generlly Environmentl Helth Perspetives, 117: llevite in gps.(4&5). Gp. `(4) showe few egenerte 5. Hrin, D., A. Peters n L. Singhl, Effets of neurons, stellitosis n neuronophgi (Fig. 6). While hroni exposure to mium, le n merury of gp.(5) revele slight improvement with moerte rin iogeni mines in the rt. Res. Commun. Chem. egenerte neurons, stellitosis n neuronophgi Pthol. Phrmol., 15: (Fig 7). Our results re prtilly greement with Bnji et l. 6. Ds, K., P. Ds, S. Dsgupt n C. Dey, [24] who reporte tht Co-supplementtion with Spirulin Serotonergi-holinergi neurotrnsrnitters' funtion restore the struturl fetures of the ereellr neurons in rin uring mium exposure in protein (improvement in the Purkinje lyer n showing restrite rt. Biologil Tre Element Reserh, restortion in the struture of Purkinje ells n ense 36: grnule lyer) in rts expose to fluorie. 7. Lfuente, A., N. Mrquez, D. Pzo n A. Esquifino, CONCLUSION Effets of suhroni lternting mium exposure on opmine turnover n plsm levels of proltin, GH n ACTH. BioMetls, 13: The results of the present stuy emonstrtes tht 8. Her, E., D. Poj-Wilzek, M. Poj, A. Pleh n Spirulin pltensis exhiite signifint protetive R. Brus, The effet of serotonin on flsh visul effet ginst mium hlorie inue neurotoxiity in evoke potentil in the rt prentlly expose to rts y inresing the enogenous ntioxint efense mium. Klinik Ozn, 103: systems in plsm n rin with susequent restortion 9. Lfuente, A. n I. Esquifino, Cmium effets of teholmine, serotonin, GABA, AChE n DNA on hypothlmi tivity n pituitry hormone frgmenttion. Furthermore, Spirulin pltensis restore seretion in the mle. Toxiol. Lett., 110: the norml histologil rhiteture of the rin. 10. Lfuente, A., N. Mrquez, M. Perez-Lorenzo, D. Pzo Aoringly it my e suggeste tht Spirulin pltensis n A. Esquifino, Puertl n postpuertl n serve s potentil therpeuti nite for the mium exposure ifferentilly ffets the rin injury ssoite with CCl 2 inue oxitive hypothlmi pituitry testiulr xis funtion in the stress in the rin. rt. Foo. Chem. Toxiol., 38: Lfuente, A., N. Mrquez, M. Perez-Lorenzo, D. Pzo REFERENCES n A. Esquifino, Cmium effets on hypothlmi pituitry testiulr xis in mle rts. 1. Usu, K., K. Kono, K. Ohnishi, S. Nkym, Exp. Biol. Me., 226: Y. Sugiur, Y. Kitmur, A. Kurit, Y. Tsu, 12. Lfuente, A. n I. Esquifino, Effets of Orl M. Kimur n Y. Yoshi, Toxiologil Cmium Exposure through Puerty on Plsm spets of mium n ouptionl helth Proltin n Gonotropin Levels n Amino Ai tivities to prevent workple exposure in Jpn: A Contents in Vrious Brin Ares in Puertl Mle nrrtive review. Toxiol In Helth, 27: Rts. NeuroToxiology, 23: Tomn, R., M. Amkoviov, S. Hluhy, M. Cj 13. Pri, L. n P. Murugvel, Dillyl tetrsulfie n J. Golin, Quntittive nlysis of the rt improves mium inue ltertions of testes fter n ute mium n izinon etylholinesterse, ATPses n oxitive stress ministrtion. Si Ppers: Animl Siene n in rin of rts. Toxiology, 234: Biotehnologies, 44: Crgeorgiou, H., V. Tzotzes, A. Sieris, A. Zrros 3. López, E., S. Figuero, M. Oset-Gsque n n S. Tskiris, Cmium effets on rin M. González, Apoptosis n nerosis: two etylholinesterse tivity n ntioxint sttus istint events inue y mium in ortil of ult rts: moultion y zin, lium n neurons in ulture, British Journl of Phrmology, L-ysteine o-ministrtion. Bsi Clin Phrmol 138: Toxiol., 97:

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