Free Serum Testosterone Level in Male Rats Treated with Tribulus Alatus Extracts

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1 Investigtive Urology Triulus Altus Extrcts nd Testosterone Level Interntionl Brz J Urol Vol. 33 (4): , July - August, 2007 Free Serum Testosterone Level in Mle Rts Treted with Triulus Altus Extrcts Wlid H. El-Tntwy, Aeer Temrz, Omym D. El-Gindi Drug Biovilility Center, Ntionl Orgniztion For Drug Control nd Reserch, Ciro, Egypt, Phrmcognosy Deprtment, Fculty of Phrmcy for Girls, Al-Azhr University, Ciro, Egypt ABSTRACT Ojective: The present study ws undertken to investigte the effect of Triulus ltus extrcts on free serum testosterone in mle rts. Mterils nd Methods: Free serum testosterone level ws mesured in mle rts treted with lcoholic extrcts of the eril prt without fruits, fruits of Triulus ltus nd their frctions. Results: All tested extrcts showed significnt increse in the level of free serum testosterone when compred to tht of corresponding control, p. Sttisticl comprison of ll groups reveled tht the mximum level ws found in groups treted with chloroformic nd ethnolic frctions of fruits extrct. Conclusion: Triulus ltus extrct ppers to possess phrodisic ctivity due to its ndrogen incresing property. Key words: Triulus; testosterone; phrodisics; rts Int Brz J Urol. 2007; 33: INTRODUCTION In trditionl medicine vriety of plnts hve een used s sex stimulnts (1). For centuries, Ars hve mde use of herl drugs to improve sexul performnce nd increse liido (2). In Africn trditionl medicine, especilly in Cmeroon, Zingier officinle nd Pentdipln-dr rzzen re used s phrodisic nd mle sexul stimultion (3). In Egypt, the pollen grins of dtes (Phoenix dctylifer) nd seeds of herml (Pegnum hrml) re used to restore sexul potency (4). The genus Triulus of the Zygophyllcee comprises c 20 species tht grow s shrus or hers in sutropicl res round the world (5).Among the Triulus species T. terrestris, T. cistoides nd T. ltus hve een phytochemiclly investigted nd isoltion of steroidl sponins from these plnts ws reported (6-8). The present study ws undertken to investigte the effect of Triulus ltus extrcts on free serum testosterone in mle rts. MATERIALS AND METHODS Plnt Mteril nd Preprtion of Extrcts Smples of Triulus ltus were collected from Al Azhr University, Nsr-city, Ciro nd were kindly identified y Deprtment of Botny, Fculty of science, Ciro University. A voucher specimen (numer 3978/1) ws deposited t Herrium Horti Botnici Pisni (Flor Aegyptic), Pis, Itly. 554

2 Triulus Altus Extrcts nd Testosterone Level The dried eril prts nd fruits (400g nd 100g, respectively) of Triulus ltus were finely powdered nd were mcerted seprtely in 70% methnol. The lcoholic extrct ws evported to dryness under vcuum. The residues were comined, weighted (90g nd 20g, respectively) suspended in distilled wter nd successively extrcted with chloroform, ethylcette nd n-utnol sturted with wter. Ech extrct ws collected nd evported to dryness under vcuum to give chloroformic extrct (4g nd 2.1g, respectively ) ethylcette extrct (3.2g nd 1.5g, respectively) nd n-utnolic extrct (8g nd 3.8g, respectively) then the wter ws evported to dryness nd the residue ws mcerted in solute ethnol severl times. The lcoholic extrcts were comined nd evported to dryness under vcuum to give ethnolic extrct (30g nd 8g, respectively) Animls Helthy, dult mle Wistr lino rts weighing g, ged 4-5 months were used in this study. The nimls hd free ccess to stndrd commercil diet, wter nd were kept in rooms mintined t 25 ± 1 C. The nimls were divided rndomly into different groups; ech group consisted of six rts. Control groups treted with distilled wter (2 ml/kg p.o.) only. Toxicity Study On the sis of the toxicity study, the LD50 vlue of the extrct of the eril prt without fruits in mice ws 812 nd ws 868 mg/kg ody weight for the extrct of fruits. On the other hnd, LD50 vlue for extrct of different frctions rnged from mg/kg ody weight. The experiment ws crried out on three steps. First Step Control group: treted with distilled wter (2 ml/kg p.o.). Group 1: received 70% lcoholic extrct of eril prt without fruits (n = 6). Group 2: received 70% lcoholic extrct of fruits (n = 6). Group 3: received 70% lcoholic extrct of totl her (n = 6). Reference group: treted with 0.45 mg mestrolone (n = 6). In the first step, groups received extrcts (suspended in wter using Tween 20 s surfctnt) t dose of 50 mg / kg ody weight orlly y orogstric ctheter once dy for 40 dys. A reference group ws treted with 0.45 mg mestrolone once dy for 40 dys orlly y orogstric ctheter. After 40 dys, lood smples were collected from the til veins of the rts t the sme time of the dy nd serum ws seprted. Second Step Control group: treted with distilled wter (2 ml/kg p.o.). Group 4: received chloroformic frction of eril prt without fruits (n = 6). Group 5: received ethylcette frction of eril prt without fruits (n = 6). Group 6: received utnolic frction of eril prt without fruits (n = 6). Group 7: received ethnolic frction of eril prt without fruits (n = 6). Reference group: treted with 0.45 mg mestrolone (n = 6). Third Step Control group: treted with distilled wter (2 ml/kg p.o.). Group 8: received chloroformic frction of fruits (n = 6). Group 9: received ethylcette frction of fruits (n = 6). Group 10: received utnolic frction of fruits (n = 6). Group 11: received ethnolic frction of fruits (n = 6). Reference group: treted with 0.45 mg mestrolone (n = 6). On the other hnd, in the second nd third steps the groups received dose 12.5 mg / kg ody weight orlly y orogstric ctheter once dy for 40 dys. A reference group ws treted with 0.45 mg mestrolone once dy for 40 dys orlly y orogstric ctheter. After 40 dys, lood smples were collected from the til veins of the rts t the sme time of the dy nd serum ws seprted. Determintion of Free Serum Testosterone The level of free serum testosterone ws mesured y Enzyme-linked immunosornt ssy (ELISA) ccording to (9), KAPD29:040318/ 2KAPD2924 IN VITRO DIAGNOSTIC USEenBioSource Europe SA - Nivelles, Belgium. Sttisticl Anlysis Dt were presented s the men ± SE (n = 6). Sttisticl nlysis used Student s t-test to compre differences etween groups nd the control. One-wy nlysis of vrince (ANOVA) ws pplied for 555

3 Triulus Altus Extrcts nd Testosterone Level comprison etween different tretments. Differences were considered sttisticlly significnt t P. RESULTS Tle-1 represents men free serum testosterone level (pg/ ml) mong group of rts treted with 70 % lcoholic extrcts of Triulus ltus. Testosterone level ws significntly incresed mong ll groups, when compred to tht of their corresponding control, P. The highest level ws found in the group treted with the reference drug followed y the group treted with fruits extrct, followed y the one treted with the eril prt without fruits nd the lowest level ws found in the group treted with totl her. Tle-2 illustrtes men free serum testosterone level (pg/ml) mong groups of rts treted with different frctions of 70% lcoholic extrcts of the eril prt without fruits of T. ltus. Testosterone level showed significnt increse mong ll groups, when compred to tht of their corresponding control, p. The level of testosterone in the group treted with the reference drug showed significnt increse when compred to tht of ll other groups. The level of testosterone in-group (5) showed significnt increse when compred to tht of other groups including tht treted y totl eril prts extrct, group (1). Tle-3 illustrtes men free serum testosterone level (pg/ml) mong groups of rts treted with different frctions of 70 % lcoholic Tle 1 Men free serum testosterone level (pg/ ml) mong groups of rts treted with 70% lcoholic extrcts of Triulus ltus. Tested Prmeter Men ± SE p Vlue Control Group 0.75 ± Group ± Group ± 0.14 Group ± c Reference Group 6.5 ± 0.98 d Vlues expressed s men ± stndrd error of 6 nimls/group. P in reltion to control group (Student s t-test). Different letters men significnce. Tle 2 Men free serum testosterone level (pg/ml) mong groups of rts treted with different frctions of 70% lcoholic extrct of eril prt without fruits of Triulus ltus. Tested Prmeter Men ± SE p Vlue Control Group 0.8 ± 0.06 Group ± 0.76 Group ± 1.02 Group ± 0.41 Group ± 0.55 Reference Group 8 ± 0.86 d Vlues expressed s men ± stndrd error of 6 nimls/group. P in reltion to control group (Student s t-test). Different letters men significnce. Tle 3 Men free serum testosterone level (pg/ml) mong groups of rts treted with different frctions of 70% lcoholic extrct of fruits of Triulus ltus. Tested Prmeter Men ± SE p Vlue Control Group 0.72 ± Group ± Group 9 8 ± Group ± Group ± 1.88 c Reference Group 7 ± 1.0 Vlues expressed s men ± stndrd error of 6 nimls/group. P in reltion to control group (Student s t-test). Different letters men significnce. 556

4 Triulus Altus Extrcts nd Testosterone Level extrcts of the fruits of T. ltus. Testosterone level showed significnt increse mong ll groups, when compred to tht of their corresponding control, p < Testosterone level showed significnt increse in groups 8 nd 11. As compred with tht of other frctions, totl fruits extrct nd the group treted with the reference drug. COMMENTS Some cuses tht re responsile for low testosterone levels, include congenitl prolems such s deficiencies of mle hormones nd rre mlformtion syndromes, nd cquired prolems such s ging, chronic illness, drugs, strvtion, stress, hed trum, infections, cncers, surgeries, lcoholism, removl or trum to the testicles, nd infection or twisting of the testicles in their sck. The use of testosterone is widespred in the tretment of mny prolems including infertility, thletic enhncement, erectile dysfunction nd liido prolems. Its ppliction cn hve grve consequences if not used properly. Androgen, or more specificlly testosterone, is widely utilized to tret erectile dysfunction (10). Vrious neurotrnsmitters nd their inter/ intrcellulr signling re responsile for the relxtion of corpus cvernosl smooth muscle. Androgens influence these neurotrnsmitters nd contriute to the regultion of penile erection. The clssic theory out testosterone tretment is tht it stimultes the sex drive nd, y doing so, restores erectile functioning (11). Once mn is dignosed s hypogondic, or hving low testosterone level, the next step is to choose which form of tretment to utilize. As with ll medictions, enefits should e evluted ginst potentil risk. Age is one importnt fctor in mking this decision. In men less thn fifty yers old, the gol is to restore liido nd erections. Testosterone lso improves strength, physicl stmin, nd helth sttus (10). Mny people re now relying on herl medicines for helth cre (12). Since other tretments pplied re ecoming more expensive nd often crry serious side effects, there should e scientific dissemintion of informtion on the therpeutic efficcy of these plnts. Aphrodisics re sustnces tht enhnce sex drive nd/or sexul plesure or cn rise sexul desire or liido (13). They re lso gents tht cn e used to modify impired sexul functions. Studies hve implicted the sponin component of plnts in enhncing phrodisic properties due to their stimultory effect of ndrogen production (11). A survey concerning the secondry metolites of genus Triulus showed tht steroidl sponins re the typicl constituents of the genus, nd in prticulr of T. terrestris (14). Sponins hve een implicted s possile ioctive gent responsile for the phrodisic effect in Triulus terrestris extrct (11). These sponins were found to increse the levels of testosterone nd luteinizing hormone (15). It ws reported tht Triulus ltus contined steroidl sponins (8), which might contriute to incresing endogenous testosterone levels y rising the level of luteinizing hormones (LH) s reported for sponins isolted from T. terrestris (15). In the present study, the significnt increse in the level of free serum testosterone is n indiction of the phrodisic potentil of Triulus ltus extrct. CONCLUSION The lcoholic extrcts of oth prts of Triulus ltus produced significnt increse in the level of free serum testosterone t dose 50 mg/kg ody weight. Also different frctions of oth prts of the plnt reveled significnt increse in the level of free serum testosterone t dose 12.5 mg/kg ody weight when compred to their corresponding controls. It is concluded tht Triulus ltus extrct ppers to possess phrodisic ctivity due to its ndrogen incresing property. CONFLICT OF INTEREST None declred. 557

5 Triulus Altus Extrcts nd Testosterone Level REFERENCES 1. Islm MW, Triq M, Ageel AM, l-sid MS, l-yhy AM: Effect of Slvi hemtodes on sexul ehviour of mle rts. J Ethnophrmcol. 1991; 33: Puri HS: Vegetle phrodisics of Indi. Q J Crude Drug Res. 1971; 11: Noumi E, Amvn ZPH, Lontsi D: Aphrodisic plnts used in Cmeroon. Fitotherpi 1998; 69: Amin ES, Awd O, El-Smd MA, Isknder MN: Phrmcologicl studies on pollen grins of dtes (Phoenix dctylefer). Phytochemistry. 1969; 8: Hegnuer R: Chemotxonomie der Pflnzen. Bsel, Birkhäuser-Verlg. 1973; 6: Mhto SB, Shu NP, Gnguly AN, Miyhr K, Kwski TJ: Steroidl glycosides of Triulus terrestris Linn. J Chem Soc Perkin Trns I. 1981; 1: Achench H, Huner H, Brndt W, Reiter M: Crdioctive steroid sponins nd other constituents from the eril prts of Triulus cistoides. Phytochemistry. 1994; 35: Temrz A, El Gindi OD, Kdry HA, De Tommsi N, Brc A: Steroidl sponins from the eril prts of Triulus ltus Del. Phytochemistry. 2006; 67: McCnn D, Kirkish L: Evlution of Free Testosterone in serum. J Clin Immunossy. 1985; 8: (ccess in August 2005). 11. Guthmn K, Adikn PG, Prsd RN: Aphrodisic properties of Triulus Terrestris extrct (Protodioscin) in norml nd cstrted rts. Life Sci. 2002; 71: Tyler VE: A textook of Hers Of Choice: The Therpeutic Use Of Phytomedicinls. Norwood, Hworth Pr Inc. 1994; P Rosen RC, Ashton AK: Prosexul drugs: empiricl sttus of the new phrodisics. Arch Sex Behv. 1993; 22: De Comrieu E, Fuzzti N, Lovti M, Merclli E: Furostnol sponins from Triulus terrestris. Fitoterpi. 2003; 74: Koumnov F, Bozdjiev E, Andreev M, Pltonv E, Ankov V: Clinicl tril of Triestn. Experiment Med. 1982; 2-4. Accepted fter revision: Decemer 23, 2006 Correspondence ddress: Dr.Wlid Hmdy Ali El-Tntwy Drug Biovilility Center Ntionl Orgniztion for Drug Control nd Reserch Dokki, P.O. 29, Ciro, Egypt Fx: E-mil: wldhmdy@yhoo.com EDITORIAL COMMENT In this mnuscript, the uthors evluted the effect of Triulus ltus extrcts on free serum testosterone in mle rts. The uthors found tht the lcoholic extrcts of oth prts of Triulus ltus produced significnt increse in the level of free se- rum testosterone t dose 50 mg/kg ody weight. Also different frctions of oth prts of the plnt reveled significnt increse in the level of free serum testosterone t dose 12.5 mg/kg ody weight. From these studies, the uthors concluded tht the lcoholic ex- 558

6 Triulus Altus Extrcts nd Testosterone Level trct of the Triulus ltus might thus e used to modify impired sexul functions, especilly those rising from hypotestosteronemi. The pper is very originl nd most prt is crefully crried out. It presents some interesting oservtions. Dr. Q. T. Yng Deprtment of Urology Second Affilited Hospitl Shntou University Medicl College Shntou, , Chin E-mil: yng2001to@126.com EDITORIAL COMMENT Mle reproductive disorders nd sexul dysfunction is serious prolem of recent society. In this respect the current pper del with topicl question of incresing incidences of mle sexul dysfunction nd potentil risk of ppliction of synthetic hormonl drugs. There is limited understnding in the literture on phrodisic effect of plnts, nmely, Triulus on mn nd primtes. The study is interesting nd informtive nd dds to our knowledge out phrodisic properties of species elonging to genus Triulus. The uthors develop proper experimentl model on rt nd provide useful dt on the wys for stimultion of ndrogen production y nturl sources. The rticle contins new fcts out isoltion of different orgnic frctions from fruits nd eril prt of Triulus nd their comprtive effect on testosterone production. The dt re useful for clinicl prctice nd tretment of mle sexul dysfunction. They will encourge further studies on finding nturl products for stimultion of testiculr endocrine function tht would e of interest for phrmcology. Dr. Nin Atnssov Inst. of Exp. Morphology & Anthropology Bulgrin Acdemy of Sciences Sofi, Bulgri E-mil: nintnssov@yhoo.com 559