International Journal of Pharmacy and Pharmaceutical Sciences

ISSN- 0975-1491 Vol 3, Issue 2, 2011

ResearchArticle

ANEVALUATIONOFDIURETICACTIVITYOFMORINDACITRIFOLIA(LINN)(NONI)FRUIT JUICEINNORMALRATS
JNANESHWARPSHENOY1,PREETHIGPAI*2,AHSANSHOEB,2PGOKUL,2AVDHOOTKULKARNI,2MSKOTIAN3
1

DepartmentofPhysiology,KSHegdeMedicalAcademy,Deralakatte,Mangalore,NitteUniversity,Karnataka,India,2Departmentof Pharmacology,KasturbaMedicalCollege,Mangalore,ManipalUniversity,Karnataka,India,3DepartmentofCommunityMedicine, KasturbaMedicalCollege,Mangalore,ManipalUniversity,Karnataka,IndiaEmail:meddocpai2@yahoo.com Received:09Dec2010,RevisedandAccepted:11Jan2011

ABSTRACT Morindacitrifolia(Nonifruit)juice,aprovenantioxidantwhenevaluatedfornephroprotectiveeffectsinamurinemodelwasnotedtoproducea highvolumeofurineformation.Henceitwasevaluatedforits diureticpotentialinnormalrats.ThestudywasconductedinsalineprimedWistar albinorats(n=6)usingfrusemide(10mg/kg)asthereferencediureticdrugwithtwooraldoses,5mg/kgand10mg/kgrespectively.Urinevolume andelectrolytes(Sodium,PotassiumandChloride)excretionwasestimatedattheendof24hoursanddatawasanalyzedbyKruskallWallisand Mann Whitney tests. P < 0.05 was considered as statistically significant. Noni fruit juice statistically increased the volume of urine (6.82 1.18ml/100g/24hr and 7.871.15ml/100gm/24hr) in a dose dependent manner increasing the diuretic index to 2.04 and 2.36 for 5ml.kg and 10ml/kgdoserangesrespectively.However,therewasastatisticalsignificantdecreaseinsodiumionexcretion(70.114.3m.mol/Lat5ml/kgand 41.979.3m.mol/Lat10ml/kg)whencomparedtothecontrol(1075.18m.mol/L).Thoughtherewasasimilardecreaseinpotassiumexcretionit wasnotstatisticallysignificant.ThesefindingsindicatethattheprobableincreaseinurineformationmightbedueanaquareticactionofNonifruit ratherthananatrureticeffectandfurtherstudieswithlargerdosesandlongerdurationarewarranted. Keywords:Aquareticactivity,Diuretic,Nonifruitjuice,Morindacitrifolia INTRODUCTION Herbal and natural products of folk medicine have been used for centuries in every culture throughout the world. Scientists and medicalprofessionalshaveshownincreasedinterestinthisfieldas theyrecognizethetruehealthbenefitsoftheseremedies.Letfood beyourmedicineandletmedicinebeyourfoodwasadvisedbythe fatherofmedicine,Hippocrates,overtwomillenniaago.Amongthe medicinal plants discovered by the ancestors of Polynesians, Morinda citrifolia L (Noni) is one of the traditional folk medicinal plantsthathavebeenusedforover2000yearsinPolynesia 1.Noni isanativeplantfromSoutheastAsiatoAustraliaandiscultivatedin Polynesia, India, the Caribbean, Central and northern South America2,3.Ithasbeenreportedtohaveabroadrangeoftherapeutic andnutritionalvalue4. Morinda citrifolia Linn (Rubiaceae), also known as Noni or Indian mulberry, Ba Ji Tian, Nono or Nonu, Cheese Fruit, and Nhau in variousculturesthroughouttheworld,isasmallevergreentree.Itis identifiable by its straight trunk, large, bright green and elliptical leaves, white tubular flowers, and its distinctive, ovoid, grenade likeyellowfruit.Thefruitcangrowinsizeupto12cmormoreand has a lumpy surface covered by polygonalshaped sections. The seeds, which are triangular shapedand reddish brown,havean air sacattachedatoneend,whichmakestheseedsbuoyant.Themature Nonifruithasafoultasteandodor5.Ithasbeenreportedtohavea broad range of health benefits for cancer, infection, arthritis, diabetes, asthma, hypertension, and pain6. Several animal studies suggestnonimayhaveanticancer7,8,immuneenhancing9andpain relievingproperties10.MostrecentlyTakashimaetal.demonstrated the medicinal uses of new constituents isolated from noni leaves11. Furthermore,ithasbeendemonstratedthatNonifruitjuicecontains someantioxidativeorantiinflammatoryingredients 12. Morinda citrifolia, has been reported to possess antithrombotic13, antioxidant14, analgesic and antiinflammatory15 and xanthine oxidase inhibitory16 activities There are also preliminary studies reporting its blood pressure lowering17 and vasodilatory18 properties.Onthedownside,reportsofserioushyperkalemiadueto its high content of potassium (56.3 meq/L), which is similar to orangeandtomatojuiceshavebeenpublished19. Since noni fruit has proven antioxidant and hepatoprotective activity(attributedtoitsantioxidantactivity),wedecidedtoexplore thenephroprotectiveeffectsofnonifruitextractinamurine model ofgentamicininducedrenaldamage.Duringthecourseofthisstudy, itwasnotedthatnonifruitjuiceadministeredratsproducedahigh volumeofurineformation.Sothisincidentalfindingpromptedusto evaluatethediureticpotentialofnonifruitjuiceinnormalrats. MATERIALSANDMETHODS Experimentalanimals AdultmaleWistaralbinorats(150200g)fromourbreedingstock wereusedforthestudy.Theywerehousedincleanandtransparent poly propylene cages with three animals in each cage and maintained at 27 with 12: 12 h lightdark cycle for a period of 7 dayspriortothestudy.Theywerefedstandardratchowandwater ad libitum. The experimental procedures described were approved bytheInstitutionalAnimalEthicsCommittee. Drugs Frusemide (Sanofi Aventis Co.) was used as a reference diuretic drug. Testdrug The fruit juice of M. citrifolia was obtained from the mature fruit grown in Karnataka. Ripe noni fruits washed and air dried, were weighed and placed in a food grade plastic container for 45 days. The noni fruit juice dripping from the pulp during this time was collectedinthecontainer,decanted(separatingthejuicefromother sediments),filtered,preservedusing10%sodiummethylparabenIP and5%sodiumpropylparabenandwasbottled.About200250ml of juice was obtained from 1kg of ripe noni fruit. After opening it waskeptinacooldryplaceandstoredat20C. Evaluationofdiureticactivity Eachanimalwasplacedinanindividualmetaboliccage24hpriorto commencementofthestudyforadaptation.ThemethodofLipschitz et al, 20,21 was employed for the assessment of diuretic activity. According to this method, the animals, deprived of food and water for 18 hours prior to the experiment, were divided into 5 groups (n=6). Group I animals received normal saline (25 ml/kg, p.o.); Group II received the standard diuretic, Frusemide (20 mg/kg, p.o.) and Groups III and IV received the test compound Noni fruit juice (5mg/kg and 10mg/kg) respectively. Before treatment, all

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IntJPharmPharmSci,Vol3,Issue2,2011,119121 the diuretic index to 2.04 and 2.36 for 5ml/kg and 10ml/kg dose ranges respectively. However it was less than that of frusemide treatedrats.(Tableo1) Table1:Effectoforaladministrationofnonifruitjuiceon urinaryvolumeexcretion Group Control Frusemide Nonifruitjuice (5mg/kg) Nonifruitjuice (10mg/kg) Urinevolume (ml/100g/24hr) 3.330.31 9.110.61* 6.821.18* 7.871.15* Diureticindex(24 hrinterval) 2.74 2.04 2.36

animalsreceivedphysiologicalsaline(0.9%NaCl)atanoraldoseof 5ml/100gbodyweighttoimposeauniformwaterandsaltload22.All thedrugswerefreshlypreparedpriortoadministration. Immediately after administration, the animals were placed in metabolic cages (each animal per cage), specially designed to separateurine andfaeces,kept at 20C0.5C.Thevolume of urine collectedwasmeasuredatthe end of 24hrs.Duringthisperiod, no food and water was made available to animals. The parameters noted were body weight before and after test period, total urine volume,andconcentrationofNa+,K+andClintheurine.Na+,K+,Cl concentrations were determined by Ion Sensitive Electrode; Roche Hitachi 917 automatic analyzer and bicarbonate ion was estimated withBloodgasanalyzer:AVLcompact3. StatisticalAnalysis TheresultswereexpressedasmeanSD.Thedatawasanalyzedby MannWhitneytestandKruskallWallistests.AvalueofPlessthan 0.05wasconsideredasstatisticallysignificant. RESULTS Effectonurinevolume Therewas noevidence of dehydrationandtheanimalswerefound normal at the observed 5hr and 24hr intervals. The reference diuretic frusemide, significantly increased the urine output when compared to control (P < 0.01), the diuretic index being 2.74. The testdrugat5and10mg/kgdoses,showedastatisticallysignificant increase in the volume of urine with a dose dependent increase in Table2:EffectoforaladministrationoftheNonifruitjuiceonurinaryelectrolyteexcretions Groups Control Frusemide Nonifruitjuice(5mg/kg) Nonifruitjuice(10mg/kg) Na+ m.mol/L 1075.18 167.817.15* 70.114.3* 41.979.3* K+ m.mol/L 5510.1 92.55.0* 50.28.9 45.332.66 Cl m.mol/L 87.335.72 132.54.42* 98.51.52* 992.61* SalureticindexNa/K Na 1.57 0.66 0.39 K 1.69 0.91 0.76 Cl 1.52 1.13 1.13 1.95 1.81 1.39 0.93

Values are expressed in meanSD; *P <0.01 compared with control group(KruskallWallisandMannWhitneytest) Diureticindex=volumeoftestgroup/volumeofcontrolgroup

Effectonurinaryelectrolyteexcretion Asindicatedintable2,thetestdrug,whencomparedtothecontrol group, showed a statistical significant decrease in the excretion of sodium, at both the dose levels tested. Though there was a similar decrease in the potassium ion excretion, it was not statistically significant.Astatisticalsignificantincreaseinchlorideexcretionwas noted in the test drug groups though it was less than that of the positive control, Frusemide which significantly increased the excretionofalltheelectrolytes.

ValuesareexpressedinmeanSEM; *P<0.01comparedwithcontrolgroup(KruskallWallisandMannWhitneytest); Salureticindex=volumeof testgroup/volumeofcontrolgroup DISCUSSION In the present study, administration of Noni fruit juice in normal Wistaralbinoratsfailedtoshowanincreaseinelectrolyteexcretion inspiteofamarkedandsignificantincreaseinthevolumeof urine. Thesefindingsindicatethattheprobableincreaseinurineformation might be due an aquaretic action of Noni fruit rather than a natrureticeffect. Although the term diuretic denotes all substances which increase urineflow(andinthissensewateritselfisadiureticagent),diuretic drugs are designed to increase sodium excretion, since cardiac edema largely results through sodium retention. In contrast, in herbal texts the term diuretic is often loosely or inaccurately applied. In particular, when a herb was taken as a decoction or infusion, as it often was traditionally, the water consumed in conjunction with the herb would have produced an observable diuresiswhichmighthavehadlittletodowithanydiureticactionof theherbitself;thiswouldpartiallyexplainvariableresultsofclinical trialswithaquareticherbs23. Those herbs which did exhibit a mild diuretic activity might have donesobecauseoftheirmineral(electrolyte)content.Confounding theissue,thetermdiureticisoftenusedinquiteadifferentcontext inherbalwritings.Herbswhicharesaidtoenhancetheexcretionof metabolic waste from the kidneys are also often described as diuretics.However,amoreaccuratedescriptionisencompassed by the terminology "diuretic depurative." Examples of diuretic depuratives include celery and clivers. Any frank diuretic action of these herbs is probably variable, depending on the individual, and unlikelytobeoutsidenormalpbysiologicallimits23. ThelatepharmacognocistVarroTyler,PhD,theorizedthatherbsact only as aquaretics, (agents that increase water excretion without affecting renal handling of electrolytes)24. In Europe, phytotherapists have proposed that the term aquaretic: more accuratelydescribessomeherbswhichgenuinelydoincreaseurine output.Thethinkinghereisthattheseherbsactontheglomerulus (unlike conventional diuretic drugs which act further along the nephron)toincreasewaterexcretionfromthebody,buttheireffect on electrolytes such as sodium and potassium is largely neutral. Aquaretics may work by causing dilation of glomerular arterioles, thereby increasing glomerular filtration rate. In other words, aquaretics act by increasing fluid loss from the body in a physiological manner, by increasing the formation of primary urine23. Various herbs like Asparagus officinalis with Petroselinum crispumhavebeenstudiedinthiscontext.Inuncontrolledtrials,this combination caused significant weight loss in overweight patients and significantly lowered blood pressure in patients with hypertension, without changing other biochemical parameters23. The distinction between an aquaretic and diuretic is critical. Aquaretics are very unlikely to affect edema or hypertension since sodium chloride is the major determinant of extracellular fluid volume and aquaretics do not influence electrolyte levels 25. They have potential for the treatment of excessive weight, hypertension, congestive heart failure, kidney stones and premenstrual syndrome23. The mineral (electrolyte) content of herbs can often underpin any observed diuretic activity. The ratio of potassium to sodium was found to be higher in decoctions of herbs which are traditionally regardedasdiuretics,comparedtootherherbs26.Apharmacological studyconcludedthatthehighpotassiumcontentofdandelionisthe 120

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agent responsible for any diuretic activity27. It is well known that potassium overloading which occurs when the kidney tubules are incapable of absorbingit,produce urinary excretion of the osmotic type28. Since noni fruit has a high content of potassium19 it can be speculated that increase in urinary volume seen during this study couldbeanosmoticeffect. Thus it can be concluded that the increase in the volume of urine observedinthisstudycouldbeduetoeitheraquareticeffectofnoni fruit juice or an ormotic effect. Since only two dose ranges were tried in this study and since the possibility of a delayed onset of action exists furtherstudieswith larger doses and moreprolonged duration are warranted. Evaluation of its activity on vasopressin receptors presents yet another portal of further investigation regarding its mechanism of action as numerous in vivo animal studies have demonstrated the unmistakable aquaretic effect of nonpeptideAVPV2receptorantagonists29. REFERENCES Whistler WA. Traditional and herbal medicine in the Cook islands.JEthnopharm1985;13:23980. 2. Dixon AR, McMillen H, Etkin NL. Ferment this: the transformation of Noni, a traditional Polynesian medicine (Morindacitrifolia,Rubiaceae).EcologicalBotany1999;53:51 68. 3. Ross IA. Medical Plants of the World. Chemical Constituents, Traditional and Modern Medical Uses Volume 2. New Jersey: HumanaPress;2001. 4. Singh Y, Ikahihifo T, Panuve M, Slatter C. Folk medicine in Tonga.Astudyontheuseofherbalmedicinesforobstetricand gynacological conditions and disorders. J Ethnopharmacol 1984;12:30525. 5. Swanholm CE, St John H, Scheuer PJ. A survey of alkaloids in Hawaiianplants.PacificScience1959;13:295305. 6. Whistler W. Tongan herbal medicine. Isle Botanica, Honolulu, Hawaii;1992.p8990. 7. McClatchey W.From Polynesianhealersto health food stores: changingperspectivesof Morindacitrifolia(Rubiaceae).Integr CancerTher2002;1:11020. 8. Hirazumi A, Furusawa E. An immunomodulatory polysacchariderichsubstancefromthefruitjuiceofNoniwith antitumouractivity.PhytotherRes1999;13:3807. 9. Hiramatsu T, Imoto M, Koyano T, Umezawa K. Induction of normal phenotypes in rastransformed cells by damnacanthal fromMorindacitrifolia.CancerLett1993;73:16166. 10. Younos C, Rolland A, Fleurentin J, Lanhers MC, Misslin R, MortierF.AnalgesicandbehavioraleffectsofMorindacitrifolia. PlantaMed1990;56:43034. 11. Takashima J, Ikeda Y, Komiyama K, Hayashi M, Kishida A, Ohsaki A. New constituents from the leaves of Morinda citrifolia.ChemPharmBull2007;55:3435. 12. Shinichi H, Wakako H, Kohei K, Toshiko S, Junichiro Y and Shogo T. Preventive effect of Morinda citrifolia fruit juice on 1. 18. 13.

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