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Ti+3( )* +,( +)>i-? Pharmacological screening of aqueous extracts of Cyperus rotundus Linn., on Obesity & bone mineral density in Wistar Rats.




Obesity is associated se!eral cancers.

ith many chronic diseases, including diabetes, heart disease, and

#he body mass index $%&'( of a )erson is 2*+,- .g/m 2 indicates o!er eight and

abo!e ,- .g/m2 re)resents obesity. World 0ealth Organi1ation $W0O( assigns obesity as global e)idemic. W0O2s latest study indicate that globally in 2--*, a))roximately ".3 billion adults $"*4( ere o!er eight and at least 5-- million adults ere obese. 6urther W0O )ro7ects that by 2-"* a))roximately 2., billion )eo)le ill be o!er eight and more than 8-- million ill be obese., 9xcess body lo %&' is associated eight and the ele!ated body mass index are strongly correlated eight is associated ith high bone mineral density $%&:(.5 ;on!ersely decrease in the body ith bone loss * and

ith an increase the ris. of fractures. 3 #he mechanism for the )ositi!e correlation

bet een obesity & %&: is unclear. <lthough increase the mechanical load on s.eletal resulting from an increase in fat and lean mass are thought to )artially ex)lain this obser!ation, 8 it is generally acce)ted that there are other factors in!ol!ed. 'ncrease adi)ose tissue may influence bone metabolism by altering hormonal factor in the circulation and through the adi)o.ines.=, > <di)onectin is the most abundant )rotein secreted from adi)ose tissue and its concentration in the circulation is much higher than that of other hormones and cyto.ines. "- #his ,-.:a )rotein circulates as !arious com)lexes and mediates its effects through binding to its s)ecific rece)tors

adi)oR" and adi)oR2 that ha!e differential tissue distribution. "" <di)onectin synthesis and secretion are de)endent on many factors and its le!els in serum adi)onectin le!els in serum are higher in omen than ith men."2 ?nli.e other adi)o.ines, serum adi)onectin le!els are decreased in conditions associated negati!ely associated

obesity and the metabolic syndrome including insulin resistance. ", @erum adi)onectin le!els are ith obesity, )articularly !isceral adi)osity."5 &oreo!er, adi)onectin le!els increase in res)onse to eight loss."* 0uman osteoblasts ha!e been re)orted to ex)ress adi)onectin and its rece)tors <di)oR" and <di)orR2 suggesting that this )rotein may be a metabolic factor lin.ing obesity to bone metabolism. "3 'n one study, treatment of human osteoblasts ith adi)onectin resulted in increased osteoblast )roliferation and differentiation, increased al.aline )hos)hatase acti!ity $<LP(, a mar.er of bone formation, and increased minerali1ation suggesting a )ositi!e effect of adi)onectin on bone."8 'n another study, adi)onectin as sho n to increase osteoclast formation by stimulating the )roduction of rece)tor acti!ator of nuclear factor+k% ligand $R<ABL( that stimulates osteoclast differentiation and acti!ity and by inhibiting the )roduction of osteo)rotegerin $OPC(, an inhibitor of osteoclastogenesis, in osteoblasts. "= #he in vivo data are conflicting. @erum adi)onectin has been re)orted to be negati!ely associated %&: at a number of s.eletal sites, in both men and addition, adi)onectin as sho n to be )ositi!ely correlated ith omen, e!en after ad7usting for fat mass. ">, 2- 'n ith serum concentrations of bone+s)ecific

<LP, and ith cross+lin.ed A+telo)e)tides of ty)e collagen $A#D(, a bone resor)tion mar.er, suggesting that adi)onectin is associated ith increased bone turno!er.2" Eurimae et al.22 similarly re)orted a hole body %&; and %&: as ell as lumbar omen, hereas some studies suggest that serum omen. 2,,25. #amura et al.2* re)orted a significant negati!e association bet een adi)onectin and s)ine %&: in both )re+and )ostmeno)ausal

adi)onectin is a )redictor of %&: in )ost+ but not )remeno)ausal the femur nec. or lumbar s)ine in )atients ith ty)e 2.

significant )ositi!e correlation bet een serum adi)onectin and %&: at the distal radius, but no effect at @e!eral studies ha!e sho n, decreases in bone density after )articular )roblem for eight loss. 23, 28 #his may be a

omen because they tend to diet more frequently than men and may ha!e more

issues related to bone health $i.e., ha!e smaller bone frames and lo er bone density(.2= Bone mass is maintained constant between puberty and menopause by the balance between osteoblast and osteoclast activity. The existence of a hormonal control of osteoblast activity has been

speculated for years by analogy to osteoclast biology. Through the search for such humoral signal(s) regulating bone formation, leptin has been identified as a strong inhibitor of bone formation. Furthermore, intracerebroventricular infusion of leptin has shown that the effect of this adipocyte-derived hormone on bone is mediated via a brain relay. Subsequent studies have led to the identification of hypothalamic groups of neurons involved in leptin's antiosteogenic function. In addition, those neurons or neuronal pathways are distinct from neurons responsible for the regulation of energy metabolism. Finally, the peripheral mediator of leptin's antiosteogenic function has been identified as the sympathetic nervous system. Sympathomimetics administered to mice decreased bone formation and bone mass. Conversely, F-blockers increased bone formation and bone mass and blunted the bone loss induced by ovariectomy.29 6rom the literature sur!ey it is quite e!ident that ;.rotundus is indicated to ha!e an effecti!e anti obesity & )roestrogenic acti!ity, besides many other )harmacological )ro)erties exhibited by the herb or its )re)arations. 't is also )ro!ed to be a safe herb. Ao study has been done so far of ;. rotundus, on its effects on obesity indicator ade)o.ine $ade)onectin(, )roestrogenic )ro)erty, Le)tin and bone mineral density / biomechanical strength of bones. 0ence the in!estigation. is en!isaged.


'n the traditional system, it as used both in its fresh and dry form. 't also mentioned in as .no n to heal ulcer and sores,dioscorides arm literature )ertaining to <rabian and Parsian medicine. 't )laster. 't as used as )erfume and an aromatic. S-i(n+i*i- n&'(? cy)erus rotundus S9n)n9'0: nut grass, musta, musta.a, nagarmotha, bhada, laldudhi, chido, mustha. G(n70: cy)erus F&'i39: cy)eraceae S>(-i(0? rotundus P&/+ 70(. ? #ubers

mention its as a diuretic in the treatment of ulcers and as an emmenagogue and an ingredient in

B)+&ni-&3 D(0-/i>+i)n? #his grass li.e herb is found in south 'ndia. 't has tubers roots or rhi1omes that are fragment. 't has been mentioned in @ahasrayoga, G )rrennial herb ith long rhi1omes, sometimes tuberous ,culms u) to 3- cm tallH lea!es 2+3 cm flo ered, the rachilla P3&n+ -,('i-&30? #he ;. rotundus ha!e been re)orted to contain oils, al.aloids, glycosides, sa)onins, fla!onoids, tannins, starch and carbohydrates. 't also contains )roteins and traces of &g, I, ;r, &n and ;o. #he rhi1ome oils of C. rotundus from 'ndia ere re)orted to ha!e J+co)aene $"".5+"2."K(, cy)erene $=.5+"".8K(, hich !alerenal $=.8+>.=K(, caryo)hyllene oxide $8.=+>.8K( and trans+)inocar!eol $*.2+8.5K(, some of ideH s)i.es o!ate, on rays to 3 cm lomngH s)i.letes linear, "+2 cm long, "2+,ingedH scales )ur)lish, carinate, obtuseH achene sub+ob!oid, trigonal, ".* mm

long, blac., minutely )a)illate.,-

ere absent in the s)ecies from other countries. #he essential oil of C. rotundus from Cermany, and found the oil to be dominated by cy)rotene, J+co)aene, cy)erene, J+selinene, rotundene, cadalene and noot.atene, among others.," U0(0? :iuretic, anthelmintic, in disorders of stomach, emmenagogue, dia)horetic, astringent, analgesic, antifungal, hy)otensi!e,, anti+inflammatory, inhibitor of )rostagland in biosynthesis and also in rhi1ome )roduced diuretic acti!ity in rats. 't is has been re)orted that the drug sho s !arious effects !i1.diabetes, antimutagens, radical sca!engers, antide)ressants, antioxidant, ound healing. #he essential oil )ossessed tranquillising acti!ity hile the ethanolic and )etroleum+ether extracts sho ed significant anti+inflammatory and anti)yretic )ro)erty. #he extracts of Cyperus rotundus inhibits acetylcholinesterase aci!ity. #he ethanol extract of aerial )arts of Cynodon dactylon and roots & rhi1omes of Cyperus rotundus sho ed mar.ed )rotection against con!ulsions induced by chemo con!ulsi!e agents in mice. #he methanol extract of nutgrass tubers also )ossessed analgesic acti!ity ele!ating the )ain threshold of mice similar to that of acetylsalicylic acid. #hera)eutic uses of Cyperus rotundus : 0igh blood )ressure ;olds and flu ;olic

;on!ulsions diarrhea :ysentery :ysmenorrheal 6e!ers 'ndigestions &alabsor)tion &eno)ause Iomiting Parasites &ental health $moodiness and de)ression( &enstrual disorders $)ain, cram)s( Agamr o7ana!anich et al.,$2--3( re)orted the inhibitory effect of hexane extract of cy)erus

rotundus and other medicinal on Aa4B4 <#Pase )um).#he hexane extract of ;.rotundus and Orthosi)hon aristatus sho ed high )otent inhibitory acti!ity on crud e en1yme Aa4,.4+<#Pase from rat brain. ,2 ?ddin et al ,$2--3( obser!ed the antidiarrhoeal acti!ity of ;.rotundus.#he methanol extract of c.rotundus rhi1ome ,gi!en orally at the doses of 2*- and *-- mg/.g body eigh,sho ed significant antidiarrhoeal acti!ity in castor oil induced diarrhoea in mice.among the fractions ,tested at 2*-mg/.g ,the )etroleum ether fraction and residuai methano fraction ere found to retain the acti!ity,the lattrr being more acti!e as com)ared to the control.#he ethyl acetate fraction did not sho antidiarrhoeal acti!ity.,, Aatara7an et al.$2--3( re)orted the antioxidant acti!ity of ;.rotundus and other medicinal )lant against free radical induction.< combination of s)eacies $)i)er nigram,).longum and 1ingiber officinale( herbs$;.rotundus and )lumbago 1eylanicam( and salts ma.es u) <mrita bindu . #hese result re!eal that <mrita bindu , a salt Ls)ice+herbal mixture exerts a )romising antioxidant )otential againsts free radical induced oxidati!e damage.,5 :uarte et al.,$2--*( re)orted the anti+candida acti!ity of ;.rotundus .,* Eagta) et al.,$2--5( obser!ed the effect of )olyherbal formulation containing ;.rotundus on any

ex)erimental models of inflammatory bo el disease. #he formulation sho ed significant inhibitory acti!ity against inflammatory bo l disease induced in these ex)erimental animal models. #he acti!ity as com)arable ith the standard drug )rednisolone. #he results obtained established the efficacy of this )oly herbal formulation against inflammatory bo el diseases.,3 0aung and Peng$2--,( obser!ed the ad!ances in the study of chemical constituents and )harmacology of C.rotundus L.,8 %amgbose et al. $2--,( studied the utili1ation of tigernut $ ;. rotundus, L.( meal in diets for coc.erel starters. #he effect of feeding graded le!els of tigernut meal $#CA( as a re)lacement for the mai1e in the diets of coc.erel starters on carcass characteristics and economics of feed con!ersion day+old chic.s as assessed for 8- days. #igernut re)laced mai1e at - K, ,,.,,K, 33.38K and "--K le!els. < total of "2ere randomly allotted to four ex)erimental diets such that each dietary treatment had ings, heads, shan.s, li!ers, hearts and ere three re)licates of ten birds. 'nclusion of #CA at ,,.,,K in coc.erel diets su))orted better carcass yield in terms of high )luc.ed, e!iscerated, drumstic., thigh, nec., lungs eights ithout significance differences $PM-.-*( in !alues obtained. 0o e!er, there

significant difference $PN-.-*( in !alues obtained. 0o e!er, there ere significant difference $PN-.-*( in bac., breast, abdominal fat, gi11ard, s)leen, .idney and intestinal eights and lengths. 'nclusion of #CA "--K le!el significantly de)ressed )arameters assessed. #he o)timum re)lacement le!el of mai1e #CA ith as ,,.,,K as this ga!e no significant reduction in carcass yield of the birds but a significant

reduction in the cost of feed consumed. 't required a feed cost of -.," ?@ dollars to )roduce one .ilogram eight gain on diet 2 $,,.,,K(. 'nclusion of #CA in the diet resulted in feed cost sa!ing of 5.==K $:2(, =."8 $:,( and =.>-K $:5( res)ecti!ely.,= 0a et al. $2--2( studied the four sesquiter)enes, beta+selinene, isocurcumenol, moot.atone and aristolone and one triter)ene, oleanolic acid ere isolated from the ethyl acetate fraction of the rhi1omes of ;. rotundus and tested for their ability to modulate gamma+ aminobutyric acid $C<%<$<(+ ben1odia1e)ine rece)tor function by radioligand binding assays using rat cerebro cortical membranes. <mong these com)ounds, only isocurunenol, one of the ne ly identified constituents of this )lants, as found to inhibit $,0(Ro"*+"8== binding and enhance $,0(flunitra1e)am binding in the )resence of C<%<. #hese results suggest that isocurcunenol may ser!e as a ben1odia1e)ine rece)tor agonist and allosterically modulate C<%<argic neurotransimission !ia enhansiment of endegenous rece)tor ligand binding.,>

@eo et al. $2--"( studied that rhi1omes of ;. rotundus ha!e been used in oriental traditional medicines for the treatment of stomach and bo el disorders, and inflammatory diseases. Aitric oxide and su)er oxide are are im)ortant mediators in the )athogenesis inflammatory diseases. #his study underta.en to addresses as eather the methanol extracts of rhi1omes of ;. rotundus could modulate AO

and O+2 )roductions by murine macro)hsge cell line, R<W 235.8 cells. #he &eO0 extract of rhi1omes of ;. rotundus sho ed the inhibition of AO )roduction in a dose+de)endent manner by R<W 235.8 cells stimulated ith interferon+gama )lus li)o)olysaccharides. #he inhibition ofAO )roduction by the extract ell as iAO@ mRA< ex)ression, determined by as due to the su))resssion of iAO@ )rotein, as

estern and northen blotting analysis, res)ecti!ely. 'n addition the &eO Of O 2+ by )horbol ester+ stimulated R<W 235.8 cells in dose+ and time L de)endent manners. ;ollecti!ely, these results suggested that the &eO0 extract of rhi1omes of C. rotundus could be de!elo)ed as anti+inflammatory candidate for the treatment of inflammatory diseases mediated by o!er)roduction of AO and O2.50sieh et al. $2---( in!estigated the ameliorating effects of the cogniti!e+enhancing ;hinese herbs administrated orally for " ee.+)anax ginseng $)g(, )anax notoginseng $PAC(, :ioscorea o))osite $:O(. Castrodia elata $C6(, sal!ia miltiorrhi1a $@&(, acorus gramineus $<C(, co)tis chinensis $;;(, Polygonum multiflorum$)m(, cy)erus rotundus $;R( and )soralea corylifolia$P;(+on the sco)olamine$@;OP(+induced amnesia by using )assi!e a!oidance tas. in rats.Often ;hinese herbs,only PC,PAC,C9 and ;; )rolonged @;OP+shortened ste)+through latency$@#L(.these results re!ealed that PC,PAC,C9 and ;; administered orally for " ee. im)ro!ed the @;OP+induced learning and memory deficit in rats.5" Eeong et al.$2---( obser!ed Rotendines <$"(,%$2(,and ;$,( the no!el sesquiter)ene al.aloids ith an un)resident carbon s.eleton, ere isolated from the rhi1omes of ;.rotendes. the structure of "+, ere elucidated by s)ectral and chemical methods.52 %amgbos et al$">>8(obser!ed the effect of feeding tagenut$;.rotundus L(meal on the )erformance of rabbits.5, #hebtaranonth et al.$">>*( )erformed acti!ity Lguided in!estigation of ;.rotendus tubers led to the isolation of )actchoulenone,cariyo)hyllene+al)ha+oxide,"-,"2+)araoxycalamene and 5,8+di methyl+ "+tetralone.#he anti+malarial acti!ity of these com)ounds are in the range of 9;*- "-$+5(+"-$+3( &, ith the no!el edo)eroxide sesquiter)ene ,"-,"2+)eroxy calamenene, exhibiting the strongest effect at 9;*- 2.,, "-$+3( &.55


Due et al.$">>5( re)orted the effect of the combination of <stragalus membranaceus$fisch( %ge. $<&(, <ngelica sinensis $oli!.( :iels $#<@( , ;y)erus rotundus L, $;R(, Ligusticum chuangxiong hort $L;( and )eaonia !eitchii Lynch $PI( on the hemorrhcological changes in Gblood stagnatingO rats.5* Due et al.$">>5( built the Gblood stagnatingO rat model 'ts hemorrhcological character ith adrenalin and cold stimulation. hole blood s)ecific

as an increase in the !iscosity , thic.ness of blood and its liability to

coagulate . the ex)erimental result sho ed that <& and #<@ could decrease the

!iscosity, but at the same time increase the )lasma s)ecific !iscosity. the Pi+regulating drug ;R and t o blood acti!ating drug L; and PI could im)ro!e the hemorrhcological changes in Gblood stagnatedO rats .the combination of qi+ regulating drugs and blood+acti!ating drugs had more fa!orable effect.5* @tudy on analgesic effect of ;.stoloniferus Ret1, stoloniferus sho ed analgesic effect in the acetic acid ;.stoloniferus in mice as "2." ml./Bg.53 @hinde et al.">== re)orted the effect of Aagarmotha $; rotundus Linn( on reser)ine induced emesis in )igeons.58 %iologically acti!e substances in subterranean )arts of )ur)le nutssedge is re)orted by 6riedman and 0oro it1 ">8" as the gro th of barley$ hordeum disichon !ar es1)eran1a( seedlings in nutrient solution as reduced by )ur)le nutsedge $cy)erus otundus( )lants gro ing in connected !ials and by addition of ethanol extract of )ur)le nutsedge, aqueous and ethanol extract from subterranean )arts of )ur)le nutsedge red seed the germination and root elongation of barley. @timulations of barley root elongation ere obtained ith diluted aqueous extract the germination of barley, mustard$ brassica as reduced in soil hich had been nigra !ar <lsace,( and cotton $Cossy)ium hirsutum !ar acala "*"8( as re)orted by I? and &ai $">>5(. #he

decocts of rhi1omes of ;.rotundus and ;.stoloniferus and total al.aloids and essential oil from ;. rithing test . the oral L: *- of essential oil of

infested )re!iously ith )ur)le nutsedge. Radical elongation of barley as inhibited by ethanol extract of soil )re!iously infested. ;hromatogra)hic studies indicated that the inhibitors are )henolic substances. Cu)ta et al.$">8"( )erformed )harmacological studies to isolate the acti!e constituents from c. rotundus )ossessing anti inflammatory, anti)yretic and analgesic acti!ities.5= Lee et al.$">8-( characteri1ed ferredoxin from nutsedge, c.rotundus L, and other s)ecies ith a high )hotosynthetic ca)acity. @ingh et al.$">8-( in!estigated )harmacological acti!ites of


;.rotundus.5> <.)eroe.o!a and abdullae! $">33( re)orted the diuretic effect of drug from and galenicals from the roots of cy)erus rotundus gro ing in a1erbaid1han. #he chemical com)osition of dry roots as as follo s: al.aloids -.2"+-.25, heart glycosides -.32+-.85 fla!onoids ".2*, )oly)henolic com)ounds ".32, saccharides before hydrolysis ",.22, saccharides after hydrolysis "5.5, starch >.2, )ectins ,.82, ethereal oils ".-3, li)id com)ounds 2.>=, resins 5.2", total acidity ex)ressed as malic acid,.2*K and !itamin c =.=mgK. the ater extract ere nontoxic for hite miceH the L:*- of the alcohol extract $2:"( as induced by as determined as >-g/.g. both the drug form and galenicals increased diuresis glycosides$"".3K( list and horhammer,$">3>+">8>(.*'ndira et al. $">*3( re)orted the occurrence of estrogenic substance in )lants. L. estrogenic acti!ity of cy)erus rotundus. #he oil of c. rotundus exhibits lo order estrogenic acti!ity. #he as hydrocarbon fraction is more acti!e than other fractional distillates, but none of the com)onents

administration of resins, al.aloids$,>.3( )oly)henolic com)ounds, fla!onoids and ethereal oils and

found as acti!e as the oil. #he )robability of these com)ounds being )roestrogens is indicated by the ratio of systemically acti!e to locally effecti!e concentration. Ao correlation exists bet een antibacterial acti!ity and estrogenic )otency of the oil and its fraction.*" #.8 OBJECTIVE OF THE STUDY

". Pre)aration of ;. rotundus aqueous extracts 2. #o induce ;afeteria diet / 0igh fat diet fed obesity, in Wistar rats of either sex and e!aluate the effect of aqueous extracts of C.rotundus against de)osition. ,. #o e!aluate the effect of aqueous extracts of ; .rotundus on %one mineral density & biomechanical strength in obese Wistar rats. 5. #o e!aluate the effect of aqueous extracts of C.rotundus on the <de)o.ine ie., <di)onectin & @erum Le)tin le!els eight gain / loss and effect on adi)ositic fat





















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