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1 with the queous extrct of Dctylorhiz mculte roots on serum leptin levels nd body weight in mle rts Hjini Mlih 1, Krgr Jhromi Hossein 1 *, Krgr Jhromi Zhr 1, Khbbz Khermeh Zhr 2 Received:10/6/2014 Revised:11/17/2014 Accepted: 12/6/ Zoonoses Reserch Center, Jhrom University of Medicl Sciences, Jhrom, Irn 2. Young Reserchers Club, Islmic Azd University, Jhrom Brnch, Jhrom, Irn Pr J Med Sci 2015;12(4):9-14 Abstrct Introduction: Obesity is chronic disese with globlly incresing incidence tht cn contribute to the development of severl diseses, such s dibetes, crdiovsculr diseses, hypertension nd some types of cncer. A wide rnge of methods hve been proposed nd dopted for treting obesity, including herbl therpy. The present study ws conducted to investigte the effect of the queous extrct of Dctylorhiz mculte roots on serum leptin levels nd body weight in mle rts. Mterils nd Methods: In the present experimentl study, 50 dult mle Wistr rts were selected nd rndomly divided into 5 groups of equl size, including negtive control group (without receiving ny substnces), the shm control group (receiving 1 ml of distilled wter), experimentl group 1 (receiving 20 mg/kg queous extrct of Dctylorhiz mculte roots), experimentl group 2 (receiving 40 mg/kg queous extrct of Dctylorhiz mculte roots) nd experimentl group 3 (receiving 80 mg/kg queous extrct of Dctylorhiz mculte roots). The extrct ws intrperitonelly injected to the experimentl groups for 28 dys. On the 29th dy, blood smples were tken from the rts for ssessing serum leptin levels. The rts' weight ws lso mesured dily throughout the experiment. Dt were nlyzed using the ANOVA nd Duncn's rnge test. Results: The dose-dependent injection of the queous extrct of Dctylorhiz mculte roots led to significnt increse in serum leptin levels nd significnt decrese in food intke nd body weight compred to the control group (P<0.05). Conclusion: The queous extrct of Dctylorhiz mculte roots reduces body weight by incresing serum leptin levels. Keywords: Dctylorhiz mculte, Leptin, Body Weight, Mle Rt Introduction Obesity is chronic disese with globlly incresing incidence tht cn led to mny diseses including dibetes, crdiovsculr diseses, hypertension, nd some cncers (1). Tody, mny different * Corresponding uthor, Address: Jhrom, University of Medicl Sciences methods re used worldwide to tret obesity such s mediction, diet, liposuction, exercise, nd physicl ctivities. Other weight control nd obesity tretment methods include metbolic Tel: Emil: hossein.krgrjhromy@gmil.com 9

2 stimultion nd incresed energy consumption using herbl substnces (2). Dctylorhiz mculte, Lncibrctet (C.koch) Renz Dctylorhiz, previously known s Mculte L. (Orchis), belongs to the orchid fmily, nd with mny different species, nd grows throughout the world. Normlly, the roots of this plnt cn be hrvested in erly summer, nd mintins its medicinl property for two yers (3, 4). This plnt contins compounds such s glucomnnn, nitrogen, strch, protein, sugr, hydroxybenzldehyde, ferulic cid, quercetin, ducosterol, cirsilineol nd steroids (5, 6). In trditionl medicine, Dctylorhiz mculte is dministered for lung diseses, intestinl disorders, tuberculosis, dirrhe, Prkinson's disese, cncer, nd fever, nd especilly to enhnce libido, tret erectile dysfunction, increse stmin nd energy. Dctylorhiz mculte is lso used in ice-crem, drinks, nd confectionry industries (7, 8). One of the min components in queous root extrct of Dctylorhiz mculte is n queous fiber clled glucomnn, with long known effects on weight loss, blood sugr control nd cholesterol reduction (9, 10). Studies show tht fibers, prticulrly queous ones, re effective in weight control through reducing bowel movement nd bsorption rtes, incresing secretion of cholecystokinin nd regultion of leptin secretion (11, 12). Receiving nd consuming energy is blnced by severl complex hormonl nd neuronl mechnisms tht ffect metbolism nd ppetite ccording to nutrient molecules in blood nd body ft. Leptin is known s weight nd ppetite controlling hormone (13). Leptin is 16kD hormone produced by ob gene tht is essentil in regulting nd reducing norml weight. Leptin is minly synthesized in white dipose tissue, nd to lesser extent, in intestinl epithelium, plcent, muscles nd the brin. The min physiologicl role of leptin is to reduce body weight nd increse energy production from body reserves (14). Since the effect of Dctylorhiz mculte on body weight nd ppetite-controlling hormones hs not been directly studied so fr, the present study imed to ssess the effects of this plnt extrct on body weight nd leptin concentrtion in mle rts. Mterils nd methods Smple collection nd extrction method: Dctylorhiz mculte ws collected from round Ysuj town in erly summer, nd its glndulr roots were clened, wshed, nd dried in shde in vitro. Using electric mill, dried smples were powdered, nd the resulting powder ws mixed with ethnol 96% t 5:1 rtio, nd thoroughly stirred in rotry t room temperture for 24 hours to obtin homogeneous solution. Next, the solution ws filtered, nd left to dry t room temperture to turn into n lcohol-free solid extrct. The solid extrct ws dissolved t dosges of 20, 40, nd 80 mg in 1 cc of double-distilled wter, nd kept refrigerted until use (15). Animls nd their grouping: All ethicl issues concerning working with lbortory nimls were observed nd registered t the ethics committee of Jhrom University of Medicl Sciences, No D/P/2991, dted In this experimentl study, 50 dult mle Wistr rts of men weight grms were used, nd initilly housed in the niml house of Jhrom University of Medicl Sciences for one week to dpt to the new environment. In the course of study, nimls were housed in 12 hourly lternte light/drk cycles t C, with free ccess to food nd wter. Animls were rndomly divided into 5 groups (10 ech): negtive control group tht received no substnces, shm group tht received intrperitonel injection of 1ml of distilled wter ccording to body weight, nd tril groups 1, 2, nd 3 tht received intrperitonel injection of 20, 40, nd 80 mg/kg body weight dily doses of queous Dctylorhiz mculte extrct respectively for 4 weeks. 10

3 leptin level (ng/m( Blood smpling nd hormone tests: By the end of the study (on the 29th dy), fter weighing, blood smples were tken from the hert using 5 cc syringes (under nesthesi using diethyl-ether). Then, blood serum ws seprted by centrifuging t 3000 rpm for 15 minutes, nd kept frozen t -20 C until testing. Leptin level ws mesured using ELISA kit (Biospes Compny, Chin) for rts. Sttisticl nlysis: Dt were nlyzed in SPSS-21 softwre using one-wy vrince nlysis, nd Duncn test (when difference between groups ws significnt) t significnce level P<0.05. Results obtined re presented s men ± SEM. Results According to results, injection of moderte (40mg/kg/dy) nd mximum (80mg/kg/ dy) doses of Dctylorhiz mculte extrct over course of 28 dys cused significnt increse in serum leptin concentrtion compred to the control group (P<0.05). However, no significnt difference in serum leptin concentrtion ws observed t minimum dose (20mg/kg/ dy) compred to the control group (figure 1). Weighing mle rts showed tht moderte nd mximum doses of Dctylorhiz mculte extrct over course of 28 dys cused significnt weight loss compred to the control group (P<0.05), but no significnt weight loss ws observed t minimum dose compred to the control group (figure 2). Therefore, with incresed concentrtion of leptin fter 28 dys of injecting moderte nd mximum doses of Dctylorhiz mculte extrct, significnt weight loss ws chieved (figures 1 nd 2) (tble 1). b c Figure 1: Chnges in serum leptin concentrtion in experimentl groups receiving different doses of Dctylorhiz mculte extrct over 28 dys compred to control group Tble 1: Chnges in serum leptin concentrtion nd body weight in experimentl groups receiving different doses of Dctylorhiz mculte extrct over 28 dys compred to control group Group/vrible Negtive control Shm Tril 1 Tril 2 Tril 3 Leptin (ng/ml) ± ± ± ± b ± c Body weight (gr) ± ± ± ± b ± bc -According to Duncn test, men vlues in ech row with t lest one common letter hve no significnt difference t 5% level 11

4 Body Weight (gr) Figure 2: Chnges in body weight in experimentl groups receiving different doses of Dctylorhiz mculte extrct over 28 dys compred to control group Discussion The present study results showed tht dose-dependent queous Dctylorhiz mculte extrct cused incresed serum leptin concentrtion, reduced food intke, nd weight loss. In this study, weight loss my be ttributed to the increse in serum leptin concentrtion due to Dctylorhiz mculte extrct. Leptin hs been shown to be involved in control of food intke through negtive fter-effect mechnism (16), by connecting to its specific receptors in hypothlmus, nd chnging expression of neuropeptides tht regulte receiving nd consuming energy, such s neuropeptide Y (NPY). Leptin is ble to directly inhibit expression of NPY tht increses food intke nd reduces energy consumption (17). It lso increses expression of CRH gene in prventriculr nucleus of hypothlmus tht is responsible for reducing ppetite (18). Leptin function is considered s obesity prevention messge, so tht obesity is observed in ob/ob nd db/db rts with leptin deficiency or resistnce (19). Administrtion of recombinnt leptin in rts cused stiety, incresed energy consumption nd weight loss (20). On the other hnd, concentrtion of leptin is indictive of stored energy in dipose tissue, nd leptin is directly involved in regulting dipose tissue metbolism b through prcrine/utocrine function with inhibition of lipogenesis nd nd stimultion of lipolysis (21). Vrious physiologicl fctors nd hormones such s cortisol, insulin, estrogen, nd glucocorticoids re involved in regulting leptin level. Evidence from in vitro studies on humn nd rt dipose tissue suggest tht insulin stimultes leptin mrna expression nd its secretion from dipose tissue (22). Studies conducted on rts indicte tht insulin increses both ob gene nd plsm leptin in helthy nd dibetic rts (23). Mechnism of insulininduced leptin secretion is s follows: insulin cuses trnsfer of glucose into dipose cells through glucose-trnsferring protein (GLUT4), nd glucose cts s n intrcellulr signl, cusing stimultion of leptin secretion from dipose cells (24). Mny compounds in Dctylorhiz mculte root ffect incresed secretion of insulin. Studies show tht dministrtion of ferulic cid in dibetic rts reduces blood sugr by incresing insulin secretion (25, 16). Quercetin mintins insulin ctive by preventing its glucosyltion (27). Thus, in this study, incresed insulin level due to Dctylorhiz mculte extrct is possible leptin-incresing mechnism. Furthermore, stimulting effects of Dctylorhiz mculte hve been bc 12

5 References: 1. Mthieu P, Poirier P, Pibrot P, et l. Viscerl obesity: the link mong inflmmtion, hypertension, nd crdiovsculr disese. Hypertension 2009; 53(4): Westerterp-Plnteng MS. Green te ctechins, cffeine nd body-weight regultion. Physiol Behv 2010; 100(1): Freudenstein J, Rsmussen FN. Sectile pollini nd reltionships in the Orchidcee.Plnt Syst Evl 1997; 205 (4): Cozzolino S, Widmer A. Orchid diversity: n evolutionry consequence of deception? Trends Ecol Evol 2005; 20(9): Bronelumg MR, Cozzolino S, Kocyn A. Exine micromorphology of Orchidine (Orchidoidee, Orchidcee): phylogenetic constrints or ecologicl influences? Ann Bot 2006; 98(1): Grieve M. A modern herbl the medicinl, culinry, cosmetic nd economic properties, cultivtion nd folk- lore of herbs, grsses, fungi, shrubs nd trees with their modern scientific uses. New York 1989: Ky S, Tekin AR. The effect of slep content on the rheologicl chrcteristics of typicl ice- crem mix. J Food Eng 2001; 47(1): Frhoosh R, Rizi A. A compositionl study on two current types of slep in Irn nd their rheologicl properties s function of concentrtion nd temperture. Food Hydrocolloids 2007; 21(3): Tekinsen KK, Guner A. Chemicl composition nd physiocochemicl proprtiies of tuber slep produced demonstrted on pituitry-testes xis, incresed testosterone, spermtogenesis nd libido, but incresed testiculr nd body weights hve not been reported. Reserchers hve ttributed this to the presence of glucomnnn in Dctylorhiz mculte (15). Glucomnnn is n queous fiber tht constitutes 7% to 61% of vrious Dctylorhiz mculte species, nd hs mjor role in weight loss, blood sugr control nd cholesterol reduction (9, 10). Studies on the effect of glucomnnn on weight loss hve reported tht it increses viscosity of gstric contents, reduces emptying rte nd bsorption of contents, increses cholecystokinin secretion, nd thus controls body weight (24). It seems glucomnnn indirectly ffects leptin level by incresing secretion of cholecystokinin. There is evidence tht in certin circumstnce, coopertion between cholecystokinin nd leptin leds to weight loss nd reduced clorie intke (28). Conclusion It ppers tht components in queous root extrct of Dctylorhiz mculte reduce food intke nd thus body weight by regulting secretion of leptin. Acknowledgement In This Wy, The Authors From The University Of Medicl Sciences And Jhrom Are Grteful For Helth Services Reserch Funding. Conflict Of Interest The uthors declred no potentil conflicts of interests with respect to the uthorship nd/or publiction of this rticle. from som Prchidce species. Food Chem 2010; 121(2): Mchessult Rh. Structurl studies on tricttes of mnnn nd glucomnnn crbohydrte. Polym 1981; 1(2): Slvin JL. Dietry fiber nd body weight.nutrition 2005;21: Keithley J, Swnson B.Glucomnnn nd Obesity: A Criticl Review.Altern Ther 2005; 11(6): Obici S. Minireview: moleculr trgets for obesity therpy in the brin.endocrinol 2009; 150(6): Wilding J PH. Leptin nd control of obesity. Curr Opin phrmcol 2001;1; Frji Z, Nikzd H, Privr K, et l. The effect of queous extrct of Slep Tubers on the structure of testis nd sexul hormones in mle mice. J Jhrom U Med Sci 2013; 11( 1): Mntzoros CS. The role of leptin nd hypothlmic neuropeptides in energy homestsis;up to dte on Leptin in obesity. Growth Horm IGF Res 2001;A Suppl: S85-S Lee MJ, Fried SK. Integrtion of hormonl nd nutrient signls tht regulte leptin synthesis nd secretion. Am J Physiol Endocrinol Metb 2009; 296(6): Trtgli LA, Dembski M, Weng X, Deng N. Identifiction nd expression cloning of leptin receptor OB-R. Cell 1995; 83: Fried SK,Ricci MR,Russell CD, et l.symposium: Adipocyte function, differentition nd metbolism.j Nutr 2000;130:3127S-31S. 13

6 20. Gordn P,Gvrilov O. The clinicl uses of leptin. Curr Opinio in phrmcol, 2003; 3: Coppck SW, Pinkney JH, Mohmmd-Ali V.Leptin production in humn dipose tissue.proc Nutr Soci 1998;57: Wbitsch M, Jensen PB, Blum WF,et l. Insulin nd cortisol promote leptin production in cultured humn ft cells. Dib 1996; 45: Velsque MT, Bhthen SJ, Hnsen CT. Leptin nd its reltion to obesity nd insulin in the SHR/Ncorpulent rt, model of type 2 dibetes mellitus. Int J Exp Dibetes Res, 2001; 2: Wng JL, Chinookoswong N, Scully S, et l. Differentil effects of leptin in regultion of tissue glucose utiliztion in vivo. Endocrinol 1999; 140: Ohnishi M, Mtuo T, Tsuno T,et l. Antioxidnt ctivity nd hypoglycemic effect of ferulic cid in STZ-induced dibetic mice nd KK-Ay mice. Biofctors 2004;21: Sri Blsubshini M, Rukkumni R, Menon VP. Protective effects of ferulic cid on hyperlipidemic dibetic rts. Act Dibetol 2003;40: Asgry S, Nderi GA, Zdegn NS, et l.the inhibitory effects of pure flvonoids on in vitro protein glycosyltion. J Herb Phrmcother 2002;2(2): Cupples WA. Regultion of body weight. Am J Physiol Regul Integr Comp Physiol 2002; 28(5):R1264-R