Aville online t www.sholrsreserhlirry.om Sholrs Reserh Lirry Der Phrmi Lettre, 217, 9 [11]: 7-15 [http://sholrsreserhlirry.om/rhive.html] ISSN 975-571 USA CODEN: DPLEB4 Effets of Chroni Consumption of Col nitid on Meltonin nd Enzyme Prodution in Alino Wistr Rts Udeuni AC, Otitoju O*, Akniru CN, Nwogu LA, Ar PN Deprtment of Biotehnology, Federl University of Tehnology, Owerri Nigeri *Corresponding uthor: Otitoju O, Deprtment of Biotehnology, Federl University of Tehnology, Owerri Nigeri. E-mil: hiufo@yhoo.om ABSTRACT The effet of Col nitid onsumption on meltonin prodution in lino mle Wistr rts ws investigted. Wistr rts weighing 15-22g were fed for 3 dys with different perentges of powdered Col nitid (5%, 1%, 2%, 3% nd % w/w). Serum onentrtions of meltonin, sori id (AA), glutthione (GSH), serum lkline phosphtse (ALP), serum sprtte minotrnsferse (AST) nd lnine minotrnsferse (ALT) of lino rts were determined using stndrd methods. Results show tht meltonin level inresed in the rnge of 14.8-23.8 ng/ml. As the onentrtion of Col nitid in the exposed groups inresed there ws derese in meltonin onentrtion (9.3 ng/ml). There ws signifint derese (p<.5) in the level of sori id (.85 to.57 mg/dl) nd glutthione (5.7 to 2.5 mg/dl) ompred to the ontrol group whih showed 8.9 mg/dl nd 6.7 mg/dl respetively. Similrly, sprtte minotrnsferse (56.7 to 24.8 U/l) nd lkline phosphtse tivities (54.5 to 25.3 Ul) deresed signifintly (p<.5) s the onentrtion of Col nitid inresed ompred to ontrol. However, lnine minotrnsferse tivity inresed s the onentrtions of Col nitid inresed etween 2 to 3%. This study shows tht Col nitid redue the levels of meltonin prodution in exposed rts nd my onsequently use sleeping relted disorders mong onsumers or exposed groups. Keywords: Col nitid, Diet, Meltonin, Asori id, Alino rts INTRODUCTION Nuts exist in nture, whih re highly onsumed s prt of mn s herished diet. However, its hemil onstituents n fvorly influene ody physiology. Col nitid is one of suh nuts tht its onsumption stimultes the entrl nervous system, gives physil strength, nd help to redue hunger nd ftigue in individuls onsuming it [1,2]. 7
Studies hve shown tht Col nitid ontins sugr, theoromine, ffeine, polyphenol nd some proximte ompounds suh s wter, sh, fts, nd protein [3-5]. The phenoli ontents of Col nitid hve lso eing reported to inlude d-tehin, 1- epitehin nd kolnin [1]. Some of these hemil onstituents re used in everge drinks, s flvor, lxtive, hert stimultor nd sedtives [4-6]. Mny professionls onsume Col nitid euse of its stimultory nd ftigue resistnt effets on them. This hs led to inresed ddition to Col nitid for individuls onsuming it. Results of the previous studies hve onsistently shown dverse effets of Col nitid onsumption to inlude inrese ontrtion of rdi musle, inresed seretion of gstri id, nd inresed uptke of gluose in the skeletl musles of dog [7-1]. Mny reserhers hve ttriuted the iologil nd physiologil effets of Col nitid to it ffeine ontent [9,1,7]. Also toxiologil studies on ffeine onsumption hve shown lots of linil symptoms, suh s; nervousness, insomni to sensory disturnes, diuresis rrhythmi, elevted respirtion nd gstrointestinl disturnes [11]. Reprodutive effets [5,12,13], psyhotive ehviorl effets [14] nd loss of ody weight [15,16], hve lso een reported. The effets of Col nitid on inhiition of enzymes hve een ttriuted to the presene of tehin, epitehin, pigenin nd nringenin [17]. However, informtion on the effet of Col nitid onsumption on the neuroendorine modultor prmeter, suh s meltonin level in humns is sre. Meltonin is mjor neuroendorine modultor with fr-rehing iologil influene over most hormonl nd ehviorl funtions of humn [18]. There re reports tht meltonin is strongly ffeted y the length of the dy, rtifiil illumintion, eletromgneti energy, exerise, nutrition, nd other ftors [19]. There is no knowledge on the effets of nuts suh s Col nitid on meltonin seretion. Therefore, the im of this study is to investigte the effet of Col nitid on the meltonin level of lino rt. MATERIALS AND METHODS Plnt mteril olletion nd preprtion Pods of Col nitid were proured from the Ekeonunw Mrket, Owerri, Imo Stte, Nigeri nd were identified y plnt txonomist t the Deprtment of Biology, Shool of Siene, Federl University of Tehnology, Owerri. The nuts were removed from the pods, ir-dried for two weeks t room temperture nd ground into fine powder using grinder. Experimentl design Thirty (3) mle Alino Wistr rts weighing etween 12 2 grms were proured from Fulty of veterinry Mediine, University of Nigeri Nsukk nd housed in the Reserh Lortory of Deprtment of Biology, Federl University of Tehnology Owerri Nigeri. The rts were housed in five stndrd ges with the sides mde of losely meshed wire guze to llow for enough ventiltion. Eh ge housed six rts for eh group under hygieni ondition t temperture (25 ± 2 C) nd n ppreile reltive humidity with 12:12 hr. light/drk yles. Animl grouping nd feed dministrtion The nimls were fed with wter nd norml feed d liitum. The five groups of different experimentl groups were fed with different onentrtions of feed mixed with ground Col nitid seed thus; the group (1) reeived norml rt feed nd len drinking wter freely. The four other groups reeived different quntities of ground kol nut mixed with norml rt how feed s follows; group two (2) five grms (5 g) of kol nut with ninety five grms (95 g) of rt feed, group three (3) reeived ten grms (1 g) of ground kol with ninety (9 g) of rt how feed, nd group four (4) reeived twenty grms (2 g) of ground kol nut 8
with eighty grms (8g) of norml rt feed, group five (5) reeived thirty grms of kol nut with seventy (7) of rt how feed in the perentge onentrtion of 5%, 1%, 2% 3% nd % respetively. Mortr nd pestle were used to lend the mixture to form the kol nut-diet. The kol nut-diet ws dministered to the four groups on dily sis for 3 dys (Tle 1). Tle 1: The tretment regime for eh group Groups Diet Reeived Perentge Quntity of Kol Nut in Diet (%) Group 1 Kol nut-diet + wter 5% Group 2 Kol nut-diet + wter 1% Group 3 Kol nut-diet + wter 2% Group 4 Kol nut-diet + wter 3% Group 5 Norml rt feed + wter % (ontrol) Colletion of lood smples Rts were nesthetized y exposing them in desitor ontining otton wool soked with hloroform for out three minutes. After tht the rt were pled on the disseting ord srified, nd refully disseted to expose the hert region. Blood smples were olleted using 5 ml hypodermi syringe nd needle nd eh lood smple olleted ws dishrged eqully into n EDTA ottle. Blood smples were llowed to lot nd entrifuged t 3 rpm for 1 mins. The serum ws seprted using miropipettes nd then used for iohemil nlysis. Biohemil nlysis Meltonin onentrtion ws determined using the method desried y Kyo nd Borilek [2]. Asori id, Alnine minotrnsferse (ALT) nd sprtte minotrnsferse (AST) nd lkline phosphtse (ALP) were determined y the method of Dunn et l., [21]. Redued glutthione level in serum ws determined y the method of Jollow et l., [22], this method is sed on the formtion of reltively stle hromophori produt on reting with sulphur hydryl ompound (GSH) with Elmn s regent. Sttistil nlysis Eh reding ws tken in triplites nd ll dt were expressed s mens nd stndrd devition nd nlyzed sttistilly using Anlysis of Vrine (ANOVA), the vlues were onsidered signifint t p.5. RESULTS The orl dministrtion of Col nitid inresed the prodution of serum meltonin of lino Wistr rt. Results otined showed tht the highest level of meltonin prodution ws oserved in lino Wistr rts in Group 4 (3% onentrtion of Col nitid in its diet). The lowest level ws otined from lino Wistr rts in ontrol, whih were not fed with Col nitid. The levels of 9
meltonin prodution in ll the groups fed with Col nitid were higher thn tht otined from the ontrol (Figure 1). The results of levels of meltonin otined from the rt fed with different onentrtions of Col nitid were signifintly different from the ontrol. 3 25 Meltonin Conentrtion (ng/ml) 2 15 1 d 5 Experimentl Groups Figure 1: Meltonin onentrtion (ng/ml) of lino Wistr rts exposed to different onentrtions of Col nitid. Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). The glutthione onentrtions showed derese in ll they lino Wistr rt fed with different onentrtions of Col nitid (Figure 2). The lowest onentrtion of glutthione ws otined from Group 4 (3% Conentrtions of Col nitid), followed y Group 3 (2% onentrtions of Col nitid). The levels of Glutthione of those fed with 2 nd 3% of Col nitid were signifintly different from those otined from 1% onentrtion (ontrol). 8 6 GSH Conentrtion (mg/dl) 4 d 2 Experimentl Groups Figure 2: Glutthione onentrtion (mg/dl) of lino Wistr rts exposed to different onentrtions of Col nitid Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). The result of onentrtion of sori id in lino Wistr rt fed with different onentrtions of Col nitid showed signifint derese. Asori id redued in the rts fed with Col nitid ompred to the ontrol. The result lso showed tht s the onentrtion of Col nitid inresed in the diet of the rt, the onentrtion of sori id deresed. The highest sori id ws otined in the ontrol (Zero % onentrtion of Col nitid) group while the lowest vlue ws otined in group 3 (2% onentrtion of Col nitid) nd group 4 (3% onentrtion of Col nitid) (Figure 3). 1
1 Asori id Con. (x 1-3 mg/ml) 8 6 4 2 Experimentl Groups Figure 3: Asori id onentrtion (mg/ml) of lino Wistr rts exposed to different onentrtions of Col nitid. Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). The result of lkline phosphtse tivity of lino rts fed with different onentrtions of Col nitid showed tht the lkline phosphtse level in rts redued s the onentrtion of Col nitid inresed in the feeds given to the rts. The highest lkline phosphtse ws otined in-group one (ontrol) while the lowest vlue ws otined from group 4 (3% onentrtion of Col nitid). The vlues otined from rts fed with Col nitid were signifintly different (p<.5) from tht of the ontrol (Figure 4). 8 6 ALP tivity (U/l) 4 2 Experimentl Groups Figure 4: Alkline phosphtse tivity (U/l) of lino Wistr rts exposed to different onentrtions of Col nitid. Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). The result of sprtte minotrnsferse levels in rt fed with different onentrtion of Col nitid is presented in Figure 5. The result showed inrese in sprtte minotrnsferse in ontrol ompred to the other groups. The lowest sprtte minotrnsferse tivities were lowest in group 3. There is no signifint differene etween the sprtte minotrnsferse levels otined in group 2 nd 3 though the onentrtion of Col nitid fed to the rts were different. 11
8 6 AST tivity (U/l) 4 2 Experimentl Groups Figure 5: Asprtte minotrnsferse tivity (U/l) of lino Wistr rts exposed to different onentrtions of Col nitid. Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). The result of lnine minotrnsferse tivity in lino Wistr rts exposed to different onentrtions of Col nitid is shown in Figure 6. There is no signifint differene etween Alnine minotrnsferse tivity in the ontrol (% onentrtion of Col nitid) nd group 1 (1% onentrtion of Col nitid). However, there were inrese in lnine minotrnsferse level in rt fed with 2%, 3% nd 4% onentrtion of Col nitid nd there ws no signifint differene in the result otined in the three groups. 25 2 ALT tivity (U/l) 15 1 5 Experimentl Groups Figure 6: Alnine minotrnsferse tivity (U/l) of lino Wistr rts exposed to different onentrtions of Col nitid. Brs re men ± stndrd devition. Brs ering different lphet letters re sttistilly signifint (p<.5). DISCUSSION The result otined showed tht meltonin level in rt in ll the groups deresed signifintly (p<.1) when ompred with the ontrol. This signifies tht hroni onsumption of Col nitid in low doses lowers the rte of meltonin. The redution in meltonin ould e ttriuted the ffeine ontent of Col nitid whih hs een reported to inhiits meltonin synthesis through 12
its intertion with denosine [23]. It ould lso e tht the ffeine hd synergisti effets with other omponents of Col nitid. The result showed the inrese level of Col nitid in diet redued signifintly meltonin levels showing tht the higher the onsumption rtes the higher the redution rte s well. Meltonin hs een reported s mjor neuroendorine modultor of nnul nd irdin iorhythms in the ody, nd hs fr-rehing iologil influene over most of the utonomi, hormonl, nd ehviorl funtions of the humn orgnism [24]. The ulk of meltonin seen in irulting lood is derived from pinel glnd regrdless of the ft tht pinel meltonin is just one of the mny sites of meltonin prodution. Glutthione level in rt in different groups signifintly deresed (p<.1) when ompred with the group mening. This ould e due to the derese in the meltonin level, whih ould hve used non-stimultory effet on glutthione, though the orgnism involved ws still le to produe the ntioxidnt even with the redution in meltonin level. Rodriguez et l., [25] reported tht meltonin ntioxidtive effet is one of the mny funtions of meltonin. Its ility to enhne the tivities of vrieties of ntioxidtive enzymes its stimultory tions on the synthesis of nother importnt intrellulr ntioxidnt, glutthione hs lso een reported [26]. The result otined, from 2% nd 3% groups showing signifint redution when ompred with the ontrol nd the redution level in meltonin, present ws suffiient to hve sustined the glutthione prodution nd hene reffirming the ft tht meltonin hs stimulting effet on glutthione [26]. Vitmin C level in the rt group showed signifint derese (p<.5) when ompred with the group, whih mens tht the derese in meltonin level hd inhiitory effet on stimultion of vitmin C in the rts. The Vitmin C level in other groups showed signifint differene when ompred with tht of the ontrol nd thus inditing tht the meltonin level in them ws not suffiient to hve sustined good level of vitmin C prodution, this finding is ontrsting with the finding of [25]. Asori id is good free rdil svenger due to its hemil properties [27]. It is lso involved in mny physiologil hnges in living orgnisms. Asori id hs een reported to ttenute hnges in the level of some enzymes (tlse nd superoxide dismutse [28]. The study lso shows tht the rt lkline phosphtse level enzyme tivities were deresed t the 3 dys of exposure of to Col nitid. Similr result ws lso reported y Ogunmefun et l. [29], when he fed lino wistr rt with kolnut. The derese in lkline phosphtse ws proof of no dmge to the liver; however it ould e ttriuted to inhiition of enzymes synthesis s result of exposure [3]. Effets of Col nitid on serum enzyme tivity, sprtte minotrnsferse nd lkline minotrnsferse inresed these enzyme levels in rt. This ould e s result of over seretion of levels of these enzymes. This is n indition of hrmful effets on the rdi or hepti tissues [31]. CONCLUSION In onlusion, the Col nitid introdued to diets fed to lino rts hd the ility to inhiit meltonin prodution whih in turn led to redution in glutthione, vitmin C, nd other ntioxidnt in the rts exposed. Disruption of the seretion of meltonin n led to predisposition to mny diseses nd ggrvtion of mny illnesses. REFERENCES 1. Umoren, E.B., Osim, E.E., nd Udoh, P.B., The omprtive effets of hroni onsumption of kol nut (Col nitid) nd ffeine diets on explortion, nxiety nd fer in Swiss white mie. Int Res J of Phrmy nd Phrmol. 211. 13
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