Comparative Study of Aqueous and Ethanolic Leaf Extracts Of Spondias Mombin on Neurobehaviour in Male Rats

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1 IOSR Journl of Phrmcy nd Biologicl Sciences (IOSR-JPBS) e-issn: 7-3. Volume, Issue (Jn. Feb. 13), PP 9-3 Comprtive Study of Aqueous nd Ethnolic Lef Extrcts Of Spondis Mombin on Neurobehviour in Mle Rts O. R Asuquo 1*, C.E Fischer 1, O. E. Mesembe 1, A. O. Igiri 1, J.I Ekom 1 Deprtment of Humn Antomy, College of Medicl Sciences, University of Clbr, Clbr. PMB 111, Nigeri. Deprtment of Physiology, College of Medicl Sciences, University of Clbr, Clbr. PMB 111, Nigeri. Abstrct: Remedies for nxiety nd fer hve been investigted through both orthodox nd trditionl medicine; this study compred the effects of chronic dministrtion of grded doses of queous nd ethnolic extrcts of Spondis mombin on nxiety nd fer in Albino Wistr rts. Fifty rts of Wistr strin (1-1g body weight) were divided into five mjor groups A-E with ech group mde up of ten rts (n=1). Spondis mombin (,, nd mg/kg) were orlly dministered for weeks before testing. The elevted plus mze ws used to ssess nxiety behviour. Following dministrtion of grded doses of queous nd ethnolic extrcts of Spondis mombin for period of four weeks, decresed nxiety nd fer in the lbino rts ws reveled (p<.). The order of the nxiolytic property following these grded dose of dministrtion ws; durtion nd frequency in open rm of the mg/kg group ws significntly higher (9. ± 37.s &. ± 1.9/min respectively) compred to control, nd other tretment groups of Spondis mombin (P<.). Durtion in closed rm (.33 ± 1.97s) ws significntly lower compred to control. Closed rms frequency with mg/kg nd mg/kg ethnol extrct were 3.3 ± 1.7/min nd 3.1 ± 1./min. The extrcts were significntly (P<.) lower compred with control (.7 ± 1./min). However, the mg/kg ethnol extrct showed lower frequency. The durtion of grooming following four weeks of tretment with mg/kg nd mg/kg ethnol extrct of Spondis mombin were. ±.1s nd 1. 1 ±.1s respectively nd these were significntly lower compred to control. Spondis mombin dose-dependently decresed nxiety nd fer with significnt decrese observed in rts dministered with ethnolic extrcts of Spondis mombin. Anxiolytic effect is shown by decrese in the frequency of grooming nd durtion of grooming, decrese in frequency of hed dipping nd stretch ttempt posture (SAP), nd lower frequency of close rm entry by the experimentl groups. Ethnolic extrct from Spondis mombin produced better behviorl effects with decresed nxiety nd fer compred to the queous extrct nd so hs potentil s n lterntive nxiolytic nd sedtive gent compred to its queous extrct. Keywords: Spondis mombin, Fer, Anxiety, Frequency, Durtion, Ethnol, Elevte plus mze. I. Introduction Neurobehviorl test is non-invsive method employed to evlute the performnce of the centrl nervous system in n individul or group with similr exposures. Herbs re n integrl prt of the development of modern civiliztion, much of the medicinl use of plnts seems to hve been observed through observtion of wild nimls, nd by tril nd error (1). Medicinl plnts re distributed worldwide, but they re most bundnt in tropicl countries (). People from developing Countries especilly Africns still resort to the use of herbs despite dvnces nd vilbility of Orthodox medicine. Herbl medicines re used for the tretment of mentl disorders in Nigeri due to its ffordbility nd the belief tht the min cuses of mentl disorders re supernturl (3,). Plnts hve been known for their medicinl properties in the centrl nervous system nd used to influence brin function (). Reports from other Continents hve lso shown the use of trditionl medicine especilly in Chin nd the United Sttes of Americ in the tretment of psychitric disorders. (,7). Spondis mombin (Sp) is one of the herbs used for the tretment of psychitric disorders in Nigeri; it belongs to the fmily Ancrdicee. Herbl preprtions of Spondis mombin re used by trditionl medicine prctitioners in the tretment of mentl disorder in Western Nigeri (). Sp is relied on for vrious herbl remedies for numerous conditions nd virtully every prt of the tree is used; from its thick corky brk, to its leves, fruits, roots to even its flowers. The leves re considered to be ntivirl, ntibcteril, nticndidl, nd ntiseptic. It is lso used in numerous microbil problems; colds nd flu, cystitis, urethritis, sore throts, herpes, yest infections, gonorrhe, eye nd er infections, nd used externlly for infected wounds, cuts, burns nd rshes. Reserches hve been crried out on Spondis mombin, nd it ws found tht most of the results correlted with its trditionl uses. It hs been reported to be non-toxic (9). Its phrmcologicl ctivities hve been reported (1-3). Recently we reported its ntigondotrophic nd ntifertility effects (-). Therefore this reserch ws imed t compring the effect of grded doses of ethnolic nd queous extrcts of Spondis mombin on behviourl prmeters such s fer nd nxiety in rts. 9 Pge

2 II. Mterils nd method Fifty mle rts of Wistr strin (1-1g body weight) purchsed from the Deprtment of Zoology, University of Clbr, Nigeri, were housed in stndrd fcility of the Deprtment of Humn Antomy, University of Clbr, Nigeri. The nimls hd ccess to rt chow from Agrofeed Nigeri Limited nd wter d libitum. The rts were divided into five min groups A-E mde up of control nd four experimentl groups. Ech group ws mde up of ten rts..1 Preprtion of ethnolic nd queous lef extrcts of Spondis mombin The plnt Spondis mombin ws identified in the botnicl grden of the University of Clbr, Nigeri nd smple deposited in the University herbrium with the voucher number MIA 313. The leves of the plnt were hrvested, wshed nd sun-dried before blending to fine powder which ws stored in cool dry plce, until required for use. 11g of dried powdered leves ws seprtely extrcted with wter nd ethnol. Aqueous infusion of Spondis mombin leves ws prepred by mixing 1mls of distilled wter upon g of powdered smple. The mixture ws llowed to stnd for 3 minutes before filtrtion. Ethnol extrct ws obtined with 1:1 ethnol/ wter rtio by sohxlet pprtus. The mixture ws then centrifuged t bout 3xg for min nd the superntnt collected. The superntnts collected were clened of prticles by suction filtrtion using Whtmnn no 1. Filter pper nd cellulose filter pper. The extrcts were subsequently concentrted to dryness in vcuo t o C using rotry evportor nd stored in dessictor. These extrctions gve percentge yield of.% nd 3% for queous nd ethnol extrcts respectively.. Herb preprtion nd tretment The queous nd ethnolic lef extrcts of Spondis mombin were reconstituted to stock concentrtion of 1mg/ml. Grded doses ccording to body weights of rts were dministered. Group A served s control (.% sline), while groups B nd C nimls received mg/kg body weight nd mg/kg body weight of the queous extrct of Spondis mombin. Groups D nd E received mg/kg body weight nd mg/kg body weight of ethnolic extrct of Spondis mombin. The extrcts were dministered orlly with the id of n orogstric tube nd lsted for dys. The lst tretment ws hours before behviourl testing for Spondis mombin..3 Behviourl ssy The elevted plus mze (EPM) test, which provides test of nxiety nd explortion ws used for this study. The elevted plus mze pprtus consists of two () open rms nd two () closed rms in the sign of plus the open rms re versive to nimls since they re open (), the closed rms provide sense of sfety due to the enclosure. The rms mesure (3x x 1 cm). The floor of the mze ws constructed from cm plnk. The sme plnk ws lso ttched to the floor of the open rms to prevent the rodent from flling off (7). Behviour scored includes frequency of open rms entry, open rm durtion, closed rm entries, close rm durtion, grooming, nd hed dipping, frequency of stretch ttend postures nd rering ().. Sttisticl nlysis Dt mong the groups with different doses of extrcts Spondis mombin ws nlysed by one- wy nlysis of vrince (ANOVA) followed by post hoc Student s t-test. Dt were presented s mens ± SEM, p- vlue less thn. ws considered sttisticlly significnt. III. Results 3.1 Comprison of durtion nd frequency of open rms entry in the elevted plus mze in rts following weeks tretment with queous nd ethnolic extrcts of Spondis mombin. The durtion in open rms following four weeks tretment with mg/kg nd mg/kg queous extrct of Spondis mombin were 1. ± 1.s nd. ±.9s respectively. These did not differ significntly when compred with control (11. ± 3.s). The durtion in the open rms following four weeks of tretment with mg/kg ethnol extrct of Spondis mombin ws 1. ± 9.1s nd this ws lso not significntly different from control. However, following tretment with mg/kg ethnol extrct of Spondis mombin, the durtion in open rm (9. ± 37.s) ws significntly higher compred to control, mg/kg, mg/kg queous extrct nd mg/kg ethnol extrct of Spondis mombin (p<.). The frequency of open rms entry with mg/kg nd mg/kg queous extrct of Spondis mombin were 1.7 ±./min nd. ±./min respectively nd did not differ significntly from the control which ws 1. ±.1/min. The frequency of open rms entry following four weeks of tretment with mg/kg ethnol extrct of Spondis mombin ws. ±./min nd this ws lso not significntly different from control. Following tretment with mg/kg ethnol extrct of Spondis mombin however, the frequency of 3 Pge

3 Open rm entry (/min) Open rm durtion (s) open rms entry (. ± 1.9/min) ws significntly higher compred to control nd mg/kg queous extrct (p<.). This is shown in figs 1 &. 1 1,b,c,d 1 mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control; b = p<. compred to mg/kg Aqueous extrct; c = p<. compred to mg/kg Aqueous extrct; d = p<. compred to mg/kg Ethnol extrct. Fig. 1: Comprison of durtion in open rms of the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin,b 3 1 mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control; b = p<. compred to mg/kg Aqueous extrct Fig. : Comprison of frequency of open rms entry in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 3. Comprison of durtion nd frequency in closed rms of elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. The durtion in closed rms following four weeks tretment with mg/kg nd mg/kg queous extrct of Spondis mombin were.7 ± 1.s nd7.17 ±.1s respectively nd did not differ significntly when compred with control (9.17 ±.s). The durtion in the closed rms following four weeks of tretment with mg/kg ethnol extrct of Spondis mombin ws.3 ± 7.1s nd this ws lso not significntly different from control. However, following tretment with mg/kg ethnol extrct of Spondis mombin, the durtion in closed rm (.33 ± 1.97s) ws significntly lower compred to control, nd mg/kg ethnol extrct of Spondis mombin (p<.). See fig 3 The frequency of closed rms entry with mg/kg nd mg/kg queous extrct of Spondis mombin were 3.3 ±./min nd 3.3 ± 1./min respectively. Closed rms frequency with mg/kg nd mg/kg ethnol extrct were 3.3 ± 1.7/min nd 3.1 ± 1./min. The extrcts were significntly (p<.) lower compred with control (.7 ± 1./min). However, the mg/kg ethnol extrct showed lower frequency. Fig. 31 Pge

4 Closed rm entry (/min) Closed rm durtion (s) ,d 7 mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control; d = p<. compred to mg/kg Ethnol extrct Fig. 3: Comprison of durtion in closed rms of the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 1 1 mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control Fig. : Comprison of frequency of closed rms entry in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 3.3 Comprison of durtion nd frequency of grooming in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. The durtion of grooming following four weeks tretment with mg/kg nd mg/kg queous extrct of Spondis mombin (1.7 ± 7.97s nd ±.7s respectively) were not significntly different compred to control (. ±.s). The durtion of grooming following four weeks of tretment with mg/kg nd mg/kg ethnol extrct of Spondis mombin were. ±.1s nd 1. 1 ±.1s respectively nd these were significntly lower compred to control. See fig. The frequency of grooming for the groups of rts treted with mg/kg nd mg/kg queous extrct of Spondis mombin were.1 ± 1./min nd 3.33 ±.7 1/min respectively. These vlues were both significntly lower when compred to control (7.3 ± 1.1/min; p<.). the frequency of grooming for the groups treted with mg/kg nd mg/kg ethnol extrct of Spondis mombin were. ±.3/min nd 1.7 ±./min respectively nd these were lso significntly lower thn control(p<.). The frequency of grooming following tretment with mg/kg ethnol extrct ws lso lower thn the mg/kg queous extrct (p<.). This decrese seemed dose dependent even though it did not significntly decrese t ll doses of tretment. Fig 3 Pge

5 Rering Freq (/min) Grooming Freq (/min) Grooming durtion (s) mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control Fig. : Comprison of durtion of grooming in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin mg/kg Aq. mg/kg Aq. mg/kg Eth.,b mg/kg Eth. = p<. compred to control; b = p<. compred to mg/kg Aqueous extrct Fig. : Comprison of frequency of grooming in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 3. Comprison of frequency of rering in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. Figure 7 shows tht the frequency of rering following tretment with both queous nd ethnol extrcts of Spondis mombin were significntly lower when compred to control (1.33 ± 1.7/min; p<.). The frequency of rering for the mg/kg nd mg/kg queous extrct of Spondis mombin were 1.33 ± 1.1 nd 9.17 ± 1.3/min respectively while tht for the mg/kg nd mg/kg ethnol extrct treted groups were 9. ± 3.9 nd. ± 1.1/min respectively. tht for control wss. ± 1.11/min mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control Fig. 7: Comprison of frequency of rering in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 33 Pge

6 SAP freq (/min) Hed dips freq (/min) 3. Comprison of frequency of hed dips nd stretch ttend postures in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin The frequency of hed dips for ll the treted groups did not differ from control. However, this ws lower in the ethnol extrct treted groups when compred to the mg/kg queous extrct group. Fig. Figure 9 shows tht the frequency of stretch ttends postures were significntly lower in ll the treted groups of nimls when compred to control. 1 1 b b mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. b = p<. compred to mg/kg Aqueous extrct Fig. : Comprison of frequency of hed dips in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin. 1 1 mg/kg Aq. mg/kg Aq. mg/kg Eth. mg/kg Eth. = p<. compred to control Fig. 9: Comprison of frequency of stretch ttend postures (SAP) in the elevted plus mze in rts following weeks tretment with queous nd ethnol extrcts of Spondis mombin IV. Discussion The elevted plus mze (EPM) is typicl test for nxiety / fer. At weeks of tretment, the frequency of open rms entry nd durtion in open rms of the EPM were significntly higher for the mg/kg ethnol extrct. The motivtion for the EPM is the open spce nd height. Rodents hve nturl version for open spce nd height (). Thus, ferful/nxious rts would spend less time in the open. The group of rts treted with mg/kg ethnol extrct ws therefore less nxious compred to their control nd lower doses of the extrct. The closed rm entry nd durtion indictes the opposite effect. The closed rm durtion is consistent with erlier results for the mg/kg extrct. Anxious rts will spend more time in the closed rms which feel sfer. Since the group of rts treted with mg/kg spent less time in the closed rm, they explored the open rms more. Grooming is displcement rection (9). Rodents introduced into novel environment would groom more s sign of nxiety nd pprehension. The frequency nd durtion of grooming ws significntly lower for the groups of rts treted with mg/kg nd mg/kg ethnol extrct. The frequency of grooming in ddition, showed dose dependent decrese in the frequency of grooming. This decrese in frequency nd durtion of grooming for these doses indicte decrese in nxiety. The frequency of stretch ttend postures is risk ssessment behviour. An increse in this behviour indictes increse hesittion to explore nd therefore incresed nxiety nd vice vers. At ll doses of tretment, the frequency of SAP ws significntly lower thn control. There ws however no significnt dose relted effect even though it ppered dose relted. 3 Pge

7 V. Conclusion The decresed nxiolytic effect of the lef extrcts of Spondis mombin my be due to its sedtive nd ntipsychotic effect reported by Ayok et l,() nd Ayok et l, (). The nxiolytic ctivities observed with Spondis mombin lef extrcts my possibly be due to the presence of flvonoids, lkloids nd crdic glycosides found in the plnt extrcts. These chemicl constituents hve been reported to be responsible for nxiolytic nd sedtive effects observed in different plnt extrcts (3-3). References [1] Huffmn M.A. Animl self-mediction nd ethno-medicine: explortion nd exploittion of the medicinl properties of plnts. Pro Nturl Society, 3; (): [] Frnsworth, N.R & Morris, R.W. Higher plnts- the sleeping gint of drug development. Americn J Phrmceut Edu, 197; 1: -. [3] Mknjuol, R. O. A. Yorub trditionl heler in psychitry. Helers concept of the nture nd etiology of mentl disorders. Africn J Med & Med Sci, 197; 1: 3-9. [] Gureje, O., Ach, R. A. & Odejide, O. A. Pthwys to psychitric cre in Ibdn, Nigeri. 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