Antidiabetic and Antihyperlipidemic Properties of Phyllanthus emblica Linn. (Euphorbiaceae) on Streptozotocin Induced Diabetic Rats

Transcription

1 Pkistn Journl of Nutrition 9 (1): 43-51, 2010 ISSN Asin Network for Sientifi Informtion, 2010 Antiieti n Antihyperlipiemi Properties of Phyllnthus emli Linn. (Euphoriee) on Streptozotoin Inue Dieti Rts Mni Krishnveni, Snkrn Mirunlini, Knn Krthishwrn n Gnesn Dhmohrn Deprtment of Biohemistry n Biotehnology, Annmli University, Annmli Ngr, Tmilnu, Ini Astrt: Dietes is known to involve oxitive stress n hnges in lipi metolism. The present stuy ws esigne to investigte the therpeuti effets of n ethnoli extrt of Phyllnthus emli fruits on ntiieti, ntioxint n lipi profile in plsm n tissues (liver n kiney) of experimentl ietes. Thirty rts were llote rnomly into 5 groups, eh of 6 rts. Group I ws te s ontrol group, group II rts were renere ieti y intrperitonel injetion of streptozotoin (40 mg/kg w), group III rts reeive Phyllnthus emli fruit ethnoli extrt (PFEet) (200 mg/kg w) y using n intrgstri tue for 45 ys, group IV rts reeive glienlmie (600 µg/kg w), group V rts given PFEet (200 mg/kg w) lone. Ethnol extrts of Phyllnthus emli fruits ws ministere orlly t oses of 200 mg/kg oy weight for 45 ys resulte in signifint reution in loo gluose n signifint inrese in plsm insulin in ieti rts. Dieti rts h elevte levels of Totl Cholesterol (TC), Very Low Density Lipoprotein-Cholesterol (VLDL-C), LDL-holesterol, Free Ftty Ais (FFA), Phospholipis (PL), Triglyeries (TG) n erese HDL-holesterol. Dieti rts fe PFEet showe signifint reution in TC, VLDL-C, LDL-C, FFA, PL, TG n n elevtion in HDL-C. In onlusion, the oservtions from this stuy show tht Phyllnthus emli hs ntiieti n its enefiil effets on lipi profile, thus it n e reommene for use s nturl supplementry herl remey in ptients suffering from ietes mellitus. Key wors: Phyllnthus emli, streptozotoin-ietes, plsm insulin, loo gluose, lipi profile INTRODUCTION Dietes mellitus is synrome hrterize y hroni hyperglyemi n isturnes of rohyrte, ft n protein metolism ssoite with solute or reltive efiieny in insulin seretion or insulin tion (Jykr n Suresh, 2003). During hyperglyemi exessive proution of free rils is elieve to e involve in mny ieti omplitions inluing ieti neuropthy in ietes mellitus (Sim n Sugimoto, 1999). Besies hyperglyemi, severl other ftors inluing yslipiemi or hyperlipiemi re involve in the evelopment of miro n mro vsulr omplitions of ietes whih re the mjor uses of moriity n eth (Bennet n Joslin s, 1998). Dietes mellitus is lso ssoite with hyperlipiemi with profoun ltertion in the onentrtion n omposition of lipi (Oetol et l., 2006). Chnges in the onentrtion of lipi with ietes ontriute to the evelopment of vsulr isese. Exessive levels of loo holesterol elerte therogenesis n lowering high loo holesterol reues the iniene of CHD (Gruny, 1986). One of the risk ftors for oronry hert isese is elevte Totl Cholesterol (TC), Low Density Lipoprotein-Cholesterol (LDL-C) n Lower High Density Lipoprotein-Cholesterol (HDL-C) (Moreno n Mitjvli, 2003; Shefer, 2002; Hu et l., 2001; Jin et l., 2007). An iel orl tretment for ietes mellitus woul e rug tht not only ontrols the glyemi level, ut lso prevents the evelopment of theroslerosis n other omplitions ssoite with ietes mellitus. A ifferent type of orl hypoglyemi gents suh s igunies n sulphonylure re ville long with insulin for the tretment of ietes mellitus (Holmn n Turner, 1991), ut hs sie effets ssoite with their uses (Kmeshwr Ro et l., 1997; Vlithn, 1998). There is growing interest in herl remeies euse of their effetiveness, miniml sie effets in n reltively low osts. In spite of the presene of known ntiieti meiines in the phrmeutil mrket, remeies from meiinl plnts re use with suess to tret this isese possily euse they re onsiere to e less toxi n free from sie effets ompre to syntheti one (Momin, 1997). Ethnootnil informtion inites tht more thn 800 plnts re use s tritionl remeies for the tretment of ietes (Pushprj et l., 2000), ut only few hve reeive sientifi srutiny. The Worl Helth Orgniztion hs reommene the evlution of the effetiveness of plnts in onitions where sfe orthoox rugs re sre (Worl Helth Orgnistion, 1980). Phyllnthus emli Linn. (Euphoriee) ommonly known s ml is n nient her use in folk meiine for the tretment of vriety of humn ilments, Corresponing Author: Snkrn Mirunlini, Deprtment of Biohemistry n Biotehnology, Annmli University, Annmli Ngr, , Tmilnu, Ini 43

2 MATERIALS AND METHODS Animls: Mle Alino Wistr rts weighing ( g) were otine from the Centrl Animl House, Deprtment of Experimentl Meiine, Rjh Muthih Meil College, Annmli University, Annmli ngr. The nimls were kept in len n ry ges with eing of py husk, expose to 12 h rk n night yle, fe with rt pellets fee (Hinustn lever lt, Bnglore, Ini) n wter liitum. The stuy ws onute in orne with the Ntionl Institute of Helth Guie for the Cre n Use of Lortory Animls (NIH Consens Sttement, 1985). Animl stuies in the work hve een performe s per the Institutionl Animl Ethil Committee of Rjh Muthih Meil College (Reg no: 465/160/1999/CPCSEA), Annmli University, Annmli ngr. Chemils: Streptozotoin (STZ) ws purhse from Sigm hemil ompny, St. Louis, Mo, USA. All other hemils n solvents use were of nlytil gre otine from E-merk n Himei, Mumi, Ini. Plnt mteril: Phyllnthus emli fruits were ollete in n roun res of Chimrm, Tmil Nu. The Plnt ws txonomilly ientifie n uthentite y Dr. V. Venkteslu, Reer, Deprtment of Botny, Annmli University. A vouher speimen ws eposite t the Herrium of Botny. Preprtion of methnoli fruit extrt of Phyllnthus emli: Aml fruits were ut into smll piees n Pk. J. Nutr., 9 (1): 43-51, 2010 prtiulrly ietes. A eotion of the fruit with stems groun into uniform power using lener. The of Tinospor orifoli is well known remey for methnoli extrt of ml ws prepre y soking 100 vrious urinry iseses (Jyweer, 1980). Aml is g of rie powere smples in 250 ml of methnol for highly nutritious n oul e n importnt ietry 12 h. The extrts re filtere y using Whtmn No. 1 soure of vitmin C, minerls n mino is. The fruit filter pper. The filtrte ws use for phytohemil pulp is eing use in severl inigenous meil sreening. preprtions ginst vriety onitions suh s hehe (Jeen n Kuttn, 1995), liver injury (De et Preliminry phytohemil nlysis: Qulittive l., 1993), theroslerosis (Thkur et l., 1998) n nlysis rrie out for P. emli fruit rue extrt ietes (Su n Kuttn, 2002). Mishr et l. (1981) showe the presene of phytohemil onstituents reporte tht ml hs hypoholesterolemi tivity. suh s flvonois, tnnins, terpenois, lklois, Extrt of Phyllnthus emli n queretin ( flvonoi rohyrtes n proteins with sene of sterois. isolte from emli) for heptoprotetive tion ws ssesse ginst pretmol inue liver mge in Chromtogrphi purifition lino rts n mie (Gulti et l., 1995). To the est of Thin lyer hromtogrphy: The mjor ompouns our knowlege, no etile investigtions h een present in the P. emli fruit extrt were ientifie y rrie out to she light on the ntihyperglyemi n using TLC/HPTLC. ntihyperlipiemi effet of P. emli in ieti rts injete with STZ. The ore im of the present stuy ws Anlytil metho to ssess the therpeuti effets of n ethnoli fruit Applition of the smple in TLC plte: 2 µl of smple extrt of P. emli on loo gluose, non-enzymi ws pplie on preote sili gel GF254 luminium ntioxints n lipi profile in plsm, liver n kiney pltes (MERCK, 8m * 7m size). Develop the plte in of STZ-inue ieti rts. The effet of P. emli Methnol: Chloroform (3:7) moile phse. Run the ethnol extrts ws ompre to glienlmie, whih moile phse up to 9 m of the plte. Remove the plte is often use s stnr rug. n ry in ir. A spot in TLC ws performe using CAMAG is snne in 2 Densitometer n linomtt IV pplitor. High Performne Thin Lyer Chromtogrphy (HPTLC): The methnol extrt ws etermine in HPTLC/ (CAMAG TLC SCANNER II). HPTLC ws performe using CAMAG HPTLC spetrophotometer provie with snner II ensitometry Linomt pplitor. Preprtion of Phyllnthus emli loholi fruit extrt for iohemil stuies: The fruits of Phyllnthus emli were rie n powere. 100 g of power ws extrte with 250 ml ethnol for 72 h. After 72 h the suspension ws filtere through fine muslin o loth n onentrte t 40±5 C. A 13% soli mteril o otine ws store t 20 C prior to experimenttion. Whenever neee, the resiul extrt ws suspene in istille wter n use for tretment. Experimentl esign: Dietes ws inue in group of rts fter n overnight fst y single intrperitonel injetion of STZ, whih ws freshly issolve in 0.1M H itrte uffer (P 4.5). The ose ws 40 mg/kg oy weight. STZ trete nimls were llowe to rink 5% gluose solution over night to overome rug inue hypoglyemi. After 48 h of STZ ministrtion, the loo gluose rnge ove mg/l were onsiere s ieti rts n use for the experiment. In the experiment, totl of 30 rts were use. Animls were rnomize n ivie into five groups of six nimls eh. 44

3 Pk. J. Nutr., 9 (1): 43-51, 2010 Group A: Control rts reeive 3% gum i. Group B: Rts were me ieti y single intrperitonel injetion of streptozotoin (40 mg/kg oy weight) with itrte uffer (ph 4.5). Group C: Dieti rts trete with Phyllnthus emli Fruit Ethnoli extrt (PFEet) (200 mg/kg oy weight) in queous solution ily using n intrgstri tue for 45 ys. Group D: Dieti rts reeive glienlmie (600 µg/kg oy weight) in queous solution ily using n intrgstri tue for 45 ys. Group E: Control rts trete with PFEet (200 mg/kg oy weight) in queous solution ily using n intrgstri tue for 45 ys. At the en of the experimentl perio, ll the rts were kept overnight fst n nesthetize using ketmine hlorie (24 mg/kg oy weight) y intrmusulr injetion n srifie y ervil epittion etween m. Bloo smples ollete in potssium oxlte/soium fluorie-ontining tues were use for the estimtion of gluose. Plsm ws seprte for the estimtion of insulin n other iohemil prmeters. Liver n kiney tissues were issete out n trnsferre to ie ol ontiners ontining 0.9%NCl n use for vrious iohemil estimtions. Biohemil investigtions: Bloo gluose ws etermine y the metho of O-toluiine using the moifie regent of (Sski et l., 1972). Reue glutthione in the plsm n tissues ws estimte y the metho of Ellmn (1959). Asori i in the plsm n tissues ws estimte y the metho of Roe n Kuether (1943). "-Toopherol in the plsm n tissues ws estimte y the metho of (Bker et l., 1980). Lipis were extrte from plsm n tissues y the metho of (Folh et l., 1957). Totl holesterol were nlyze y the metho of (Allin et l., 1974). Triylglyerol in the plsm n tissues were estimte using the ignosti kit se on the enzymi metho esrie y (MGown et l., 1983). Free ftty is n phospholipis were estimte y the metho of (Flholt et l., 1973) n Zilversmit n Dvis (1950). HDL-holesterol ws estimte using the ignosti kit se on the enzymi metho esrie y (Izzo et l., 1981). Very Low Density Lipoprotein Cholesterol (VLDL- C) n Low Density Lipoprotein Cholesterol (LDL-C) frtions were lulte s VLDL-C = TG/5 n LDL-C = totl holesterol - (HDL-C + VLDL-C), respetively. Dt evlution: Sttistil nlysis ws performe using SPSS softwre pkge, version The vlues were nlyse y One Wy Anlysis Of Vrine (ANOVA) followe y Dunn s Multiple Rnge Test (DMRT). All these results were expresse s men±sd for six rts in eh group: p-vlues <0.05 were onsiere s signifint. RESULTS The qulittive nlysis of methnoli rue extrts of P. emli n the results were summrize in Tle 1. It shows tht flvonois, tnnins, rohyrtes, lklois, Proteins, terpenois were present in extrt. Sterios were sent in P. emli extrt. The methnol extrt of P. emli fruit shows two mjor peks. This inites tht there re two mjor ompouns whih my e present in the P. emli fruit extrt n it is shown in Tle 2. Figure 1 shows HPTLC hromtogrm of P. emli. Tle 1: Qulittive nlysis on phytohemil onstituents in the methnol extrt of P. emli fruit Flvonois + Tnnins + Crohyrtes + Alklois + Proteins + Sterois - Terpenois + + = Presene; - = Asene Tle 2: Anlysis of methnol extrt of P. emli fruit y HPTLC Pek Retention Ftor (R f) Are (%) The methnol extrt of P.emli fruit shows 2 mjor peks. This inites tht there re two mjor ompouns my e present in the P.emli fruit extrt Fig. 1: HPTLC hromtogrm of P. emli The oy weight hnges in ontrol n experimentl groups were illustrte in Tle 3. The oy weight of ieti rts were signifintly erese (p<0.05) when ompre with ontrol group. Supplementtion of PFEet n glienlmie showe signifint improvement in the oy weight of ieti rts. There were no signifint hnges oserve etween ontrol (group I) n ontrol trete group (group V). 45

4 Pk. J. Nutr., 9 (1): 43-51, 2010 Tle 3: Effet of PFEet on the hnges of oy weight of ontrol n experimentl rts Boy weight (g) Groups Initil (0 y) Finl (45 ys) Control (3% gum i) ± ±8.08 Dieti ontrol ± ±6.46 Dieti + PFEet (200 mg/kg w) ± ±7.42 Dieti + glienlmie (600 µg/kg w) ± ±9.08 Control + PFEet (200 mg/kg w) ± ±5.70 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 4: Effet of PFEet on the levels of loo gluose, plsm insulin in ontrol n experimentl rts Groups Bloo gluose (mg/l) Plsm insulin (µu/ml) Control (3% gum i) 80.52± ±1.50 Dieti ontrol ± ±0.54 Dieti + PFEet (200 mg/kg w) ± ±0.61 Dieti + glienlmie (600 µg/kg w) 91.48± ±0.79 Control + PFEet (200 mg/kg w) 82.08± ±0.79 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 5: Effet of PFEet on non-enzymti ntioxints in plsm of ontrol n experimentl rts Plsm Groups GSH (mg/l) Vitmin C (mg/l) Vitmin E (mg/l) Control (3% gum i) 24.94± ± ±0.12 Dieti ontrol 14.82± ± ±0.06 Dieti + PFEet (200 mg/kg w), 22.32± ± ±0.05 Dieti + glienlmie (600 µg/kg w) 20.71± ± ±0.03 Control + PFEet (200 mg/kg w) 22.07± ± ±0.14 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 6: Effet of PFEet on non-enzymti ntioxints in liver of ontrol n experimentl rts Liver GSH Vitmin C Vitmin E Groups (mg/100 g wet tissue) (mg/100 g wet tissue) (mg/100 g wet tissue) Control (3% gum i) ± ± ±0.21 Dieti ontrol 46.26± ± ±0.39 Dieti + PFEet (200 mg/kg w) ± ± ±0.31 Dieti + glienlmie (600 µg/kg w), ± ± ±0.27 Control + PFEet (200 mg/kg w) ± ± ±0.40 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 7: Effet of PFEet on non-enzymti ntioxints in kiney of ontrol n experimentl rts Kiney GSH Vitmin C Vitmin E Groups (mg/100 g wet tissue) (mg/100 g wet tissue) (mg/100 g wet tissue) Control (3% gum i) ± ± ±0.10 Dieti ontrol 47.32± ± ±0.15 Dieti + PFEet (200 mg/kg w) 95.53± ± ±0.34 Dieti + glienlmie (600 µg/kg w), ± ± ±0.20 Control + PFEet (200 mg/kg w) ± ± ±0.21 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) To evlute the effets of PFEet on loo gluose n plsm insulin levels of ontrol n ieti rts were summrize in Tle 4. There ws signifint (p<0.05) inrese in the loo gluose n erese in plsm insulin levels in ieti rts s ompre to ontrol. After ministrtion of PFEet n glienlmie to ieti rts were signifintly reue loo gluose n elevte plsm insulin levels. No vritions were oserve etween ontrol (group I) n ontrol trete group (group V). Tle 5, 6 n 7 shows the levels of non-enzymti ntioxints suh s, vitmin C, vitmin E n GSH in the plsm n tissues of ontrol n STZ-ieti rts. The levels of non-enzymti ntioxints were foun to 46

5 Pk. J. Nutr., 9 (1): 43-51, 2010 Tle 8: Effet of PFEet on the levels of totl holesterol in plsm n tissues of ontrol n experimentl rts Totl holesterol Plsm (mg/l) Liver Kiney Totl (mg/100 g (mg/100 g Groups Cholesterol HDL-C LDL-C VLDL-C tissue) tissue) Control (3% gum i) 76.32± ± ± ± ± ±0.21 Dieti ontrol ± ± ± ± ± ±0.56 Dieti + PFEet 94.32± ± ± ± ± ±0.68 (200 mg/kg w) Dieti + glienlmie, 83.21± ± ± ±0.09, 4.05± ±0.43 (600 µg/kg w) Control + PFEet 72.98± ± ± ± ± ±0.29 (200 mg/kg w) Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 9: Effet of PFEet on the levels of phospholipis in plsm n tissues of ontrol n experimentl rts Phospholipis Groups Plsm (mg/l) Liver (mg/100 g tissue) Kiney (mg/100 g tissue) Control (3% gum i) 78.10± ± ±1.57 Dieti ontrol ± ± ±3.02 Dieti + PFEet (200 mg/kg w) ± ± ±1.39 Dieti + glienlmie (600 µg/kg w) 80.43± ± ±1.68 Control + PFEet (200 mg/kg w) 76.53± ± ±1.24 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 10: Effet of PFEet on the levels of triglyerie in plsm n tissues of ontrol n experimentl rts Triglyerie Groups Plsm (mg/l) Liver (mg/100 g tissue) Kiney (mg/100 g tissue) Control (3% gum i) 55.45± ± ±0.38 Dieti ontrol 157.8± ± ±0.32 Dieti + PFEet (200 mg/kg w) ± ± ±0.32 Dieti + glienlmie (600 µg/kg w) 60.45± ± ±0.28 Control + PFEet (200 mg/kg w) 57.34± ± ±0.76 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) Tle 11: Effet of PFEet on the levels of free ftty i in plsm n tissues of ontrol n experimentl rts Free ftty i Groups Plsm (mg/l) Liver (mg/100 g tissue) Kiney (mg/100 g tissue) Control(3% gum i) 54.76± ± ±0.23 Dieti ontrol ± ± ±0.27 Dieti + PFEet (200 mg/kg w) 69.54± ± ±0.61 Dieti + glienlmie (600 µg/kg w) 60.57± ± ±0.14 Control + PFEet (200 mg/kg w) 56.87± ± ±0.26 Vlues re given s men±sd (n = 6 rts); Vlues not shring ommon supersript letter iffer signifintly t p<0.05 (DMRT) levels were foun to e inrese signifintly (p<0.05), the level of High-Density Lipoprotein Cholesterol (HDL- C) ws signifintly (p<0.05) erese. Aministrtion of PFEet n glienlmie to ieti rts erese the TC, LDL-C n VLDL-C levels n signifintly inrese the levels of HDL-C. No signifint hnges were oserve in the non-ieti n non-ieti rts trete with PFEet. Influene of PFEet on PL, TG n FFA in plsm n tissues of ontrol n experimentl rts were presente in Tle 9, 10 n 11. The levels of phospholipi, free ftty i n triglyerie inrese signifintly (p<0.05) e signifintly (p<0.05) erese in the plsm n tissues of ieti rts. Orl ministrtion of PFEet n glienlmie for 45 ys resulte in the ner normliztion of the levels of vitmin C, vitmin E n GSH in the plsm, liver n kiney of ieti rts. No signifint hnges were oserve etween ontrol (group I) n ontrol trete group (group V). The results of the effets of PFEet on plsm n tissue totl holesterol of ontrol n ieti rts were epite in Tle 8. In ieti rts, while the levels of TC, Low-Density Lipoprotein Cholesterol (LDL-C) n very Low-Density Lipoprotein-Cholesterol (VLDL-C) 47

6 Pk. J. Nutr., 9 (1): 43-51, 2010 in plsm n tissues of ieti rts s ompre to nti-ieti tivity (Su et l., 2004). Inihiite "- those of ontrol rts. On the progression of tretment gluosise enzyme shows reue loo gluose with PFEet n glienlmie signifintly erese level ue to the polyphenoli ompouns suh s these prmeters in ieti rts. The ontrol (group I) tnnins n sponins (Chkrvrthy et l., 1982). On the n ontrol trete group (group V) i not show ny sis of the ove eviene, it is possile tht the signifint hnges. flvonois n tnnins present in this fruit my e responsile for the oserve ntiieti tivity. Free-ril inue oxitive stress hs een implite in the pthogenesis of wie vriety of linil isorers, resulting usully from efiieny of nturl ntioxint efenses. The non-enzymti ntioxint efense systems re the seon line of efense ginst free ril mge. Vitmin C, potent wter solule non-enzymi ntioxint effetively interept oxints in the queous phse efore they ttk n use etetle oxitive mge (Beter, 1994). Vitmin C plys n importnt role in the etoxifition of retive intermeites proue y DISCUSSION Streptozotoin inue hyperglyemi in roents is onsiere to e goo preliminry sreening moel. Streptozotoin is well known for its seletive pnreti ß-ell ytotoxiity n hs een extensively use to inue Type-1 ietes in experimentl rt moel. It interferes with ellulr metoli oxitive mehnisms (Ppio et l., 2000). STZ-inue ietes is hrterize y severe loss in oy weight of untrete rts. The hrteristi loss of oy weight in ieti rts oul e ue to egrtion n tolism of fts n proteins (Hkim et l., 1997). Inrese toli retions leing to musle wsting my e the use for weight loss in ieti rts (Rjkumr et l., 1991). In our stuy, the orl ministrtion of PFEet n glienlmie normlize the oy weight in ieti rts. This suggests tht the protetive effet of the extrt in ontrolling musle wsting is glyolysis. Phyllnthus emli is lime to e useful in ietes in folklore meiines, the results inites tht the plnt extrt ws foun to reue the loo gluose level in STZ-inue ieti rts. The oserve signifint inrese in loo gluose ws ompnie y signifint erese in plsm insulin levels in ieti rts when ompre with ontrol rts. Our oservtions re in omplete greement with the reports y severl workers tht STZ-inue ietes mellitus n insulin efiieny le to inrese loo gluose (Chue et l., 2001). This oul e ue to the result of impirment of peripherl tissues of liver to metolize gluose. In the present stuy, ieti rts trete with PFEet n glienlmie showe signifint erese in loo gluose with signifint inrese in plsm insulin level s ompre to ieti ontrol rts. The possile mehnism y whih PFEet ring out its ntihyperglyemi tion my e y inresing the pnreti seretion of insulin from remnnt ß-ells, whih ws lerly eviene y the inrese level of insulin in ieti trete rts. A numer of other plnts hve een reporte to exert hypoglyemi tivity through insulin relese-stimultory effets (Twij n Al-Br, 1988). The effet proue y the extrt ws ompre with stnr rug, glienlmie. Phytohemil nlysis of the fruit revele the presene of tnnins, flvonois, lklois, terpenois, rohyrtes n proteins. These ompouns re powerful ntioxints to svenge the free rils inue y hyperglyemi. Herl extrts ontining flvonois n tnnins were reporte to emonstrte ytp whih etoxifies xenoiotis. In our investigtion, 450 the erese in vitmin C level my e ue to inrese utiliztion of vitmin C s n ntioxint efense ginst retive oxygen speies or to erese in GSH level, sine GSH is require for reyling of vitmin C (Inofers n Sies, 1988). The oserve erese in the levels of vitmin C in the ieti onition is onsistent with previous reports (Prine n Menon, 1999). Vitmin E is n importnt ril svenging ntioxint tht interrupts the hin retion of LPO y reting with lipi peroxyl ril (Grry n Buethner, 1993). In our experiment, there is erese level of vitmin E in plsm n tissues of ieti rts were notie. Reue vitmin E levels my e ue to the erese vitmin C level, euse there is well estlishe synergism etween vitmin C n vitmin E. Glutthione is one of the most unnt tripeptie, nonenzymti ntioxint present in liver. It removes free ril speies suh s hyrogen peroxie, superoxie rils n mintins memrne protein thiols. Moreover, it is sustrte for glutthione peroxise (Prksh et l., 2001). The levels of GSH in plsm n tissues ereses uring ietes represent the inrese utiliztion ue to oxitive stress. Tretment with PFEet n glienlmie showe normliztion of enzymi n non-enzymi ntioxint levels, whih suggests the effiy of extrt to svenge the retive oxygen speies overproution uring ietes. The ntioxint potentil of P. emli whih my e ue to the presene of phenoli ompouns n vitmin C. It is reporte tht phenoli ompouns in plnts possess strong ntioxint tivity n my help to protet ells ginst the oxitive mge use y free rils (Kirkosyn et l., 2003). Lipis ply n importnt role in the pthogenesis of ietes mellitus. The mrke hyperlipiemi tht hrterizes the ieti stte my therefore e regre s onsequene of the uninhiite tions of 48

7 Pk. J. Nutr., 9 (1): 43-51, 2010 lipolyti hormones on the ft epot (Csigli n onentrtion ws remrkly inrese in plsm n Pltini, 1998). In our stuy, we hve notie tht the tissues of ieti rts whih is lso ssoite with n elevte levels of serum lipis suh s Totl inrese in LDL-C, VLDL-C n erese in HDL-C. In Cholesterol (TC) n Triglyerie (TG) in plsm n ieti stte, plsm n tissue holesterol levels tissues of ieti rts when ompre with tht of were elevte ue to lk of insulin, s the tivity of ontrol nimls. This oul e ue to insulin efiieny, HMG CoA s lowere in insulin efiieny. The whih results in filure to tivte the lipoprotein lipse inrese onentrtion of holesterol oul e ue to therey using hypertriglyeriemi (Shirwkr et l., erese in HDL-C, Sine HDL is known to e 2004). Uner norml irumstnes, insulin tivtes involve in the trnsport of holesterol from tissues to the enzyme lipoprotein lipse n hyrolyses the liver for its tolism. The inrese level of Triglyeries (TG) (Suresh Kumr n Menon, 1992; plsm n tissue holesterol oserve in our stuy is Tskinen, 1987). The inrese levels of TG n TC in greement with the previous finings (Kweiterovih, in ieti rts re in greement with other reports 2000). Aministrtion of PFEet n glienlmie (Tunli n Ynrg, 2006). Upon ministrtion of ttenute signifint reution in totl holesterol, PFEet n glienlmie reue triglyerie in LDL-C, VLDL-C n signifint inrese in HDL-C in plsm n tissues of ieti rts, this my e ue to ieti nimls. low tivity of holesterol iosynthesis enzymes or low level of lipolysis tht re uner the ontrol of insulin Conlusion: Our present investigtion shows tht P. (Shrm et l., 2003). emli fruit extrt possesses ntiieti, ntioxint Fttyis, n importnt omponent of ellulr n ntihyperlipiemi effets in streptozotoinmemrnes re eiosnoi preursors n re inue ieti rts. Therefore, P.emli fruits show therefore require for oth the struture n funtion of therpeuti promise s protetive gent ginst the every ell in the oy (Rjsekrn et l., 2006). Aprt evelopment n progression of theroslerosis n from the regultion of rohyrte metolism, Insulin possile relte riovsulr omplitions in lso plys n importnt role in the metolism of lipis. ietes mellitus. It s worth emphsing tht P.emli Insulin is potent inhiitor of lipolysis. Sine it inhiits hs onsierle potentil for improving puli helth if the tivity of the Hormone Sensitive Lipse (HSL) in use on regulr sis. Further phrmologil n ipose tissues n suppresses the relese of Free iohemil investigtions re unerwy to fin out the Ftty Ais (FFA) (Cohn n Roth, 1996). In our stuy it tive onstituent responsile for the ntiieti tivity is ler tht the plsm n tissues of the ieti n to eluite its mehnism of tion. niml shows inrese level of free ftty is. Inrese FFA in ieti rts is minly ue to n inrese in the moiliztion of free ftty i from the peripherl ft epots, sine insulin inhiits the hormone sensitive lipse (Goomn n Gilmn, 1985). Our ACKNOWLEDGEMENTS The uthors knowlege the University Grnts Commission, New Delhi, Rjiv Gnhi Fellowship for proviing Junior Reserh Fellowship (JRF). results re in or with other reports (Krishnkumr et l., 2000). A signifint reution of FFA ws oserve in plsm n tissues of ieti rts fe with PFEet n glienlmie. Phospholipis (PL) re vitl prt of iomemrne rih in Polyunsturte Ftty Ai (PUFA), whih re o o suseptile for free rils suh s O 2 n OH rils (Ahme et l., 2001). Our present results inite tht the inrese levels of phospholipis in ieti rts my e ue to the elevte levels of free ftty i n totl holesterol. The phospholipis level tens to k towrs norml fter tretment with PFEet n glienlmie. Cholesterol is powerful risk ftor for mny oronry hert iseses. The egree of hyperholesterolemi is iretly proportionl to the severity of ietes (Zvroni et l., 1989). ß-Hyroxy-ß-Methylglutryl Coenzyme A (HMGCoA) reutse tlyzes the rte-limiting step in holesterol iosynthesis n its tivity orreltes losely with the rte of tissue holesterol synthesis. Aoring to our stuy, the totl holesterol REFERENCES Ahme, I., M.S. Lkhni, A. Gillet John n H. Rz, Hypotriglyeriemi n inue hypoholesterolemi effets of ntiieti Momori hrnti Krel fruit extrt in STZieti rts. Di. Res. Clin. Prt., 51: Allin, C.C., L.S. Poon, C.S.G. Chn, W. Rihmon n P.C. Fu, Enzymti of totl etermintion serum holesterol. Clin. Chem., 20: Bker, H., O. Frnk, B. DeAngelis n S. Reingol, Plsm toopherol in mn t vrious times fter ingesting free or etylte toopherol. Nutr. Res., 21: Bennet, P.H. n Joslin s, Dietes Mellitus Khn R.C, Weir GC Bltimore Interntionl pulition. Wverley, pp: Beter, R.E., The role of sorte in ntioxint protetion of iomemrne: intertion with vitmin E n CoenzymeQ. J. Bioenerg. Biomemr., 26:

8 Pk. J. Nutr., 9 (1): 43-51, 2010 Csigli, E. n P. Pltini, Criovsulr risk Izzo, C., F. Grillo n E. Muror, Improve metho ftors in the elerly. J. Hum. Hypertens., 12: 575- for the etermintion of high ensity lipoprotein 581. holesterol I. Isoltion of high-ensity lipoproteins Chkrvrthy, B.K., S. Gupt n K.D. Goe, y the use of polyethylene glyol Clin. Chem., Funtionl ß-ell regenertion in the islets of 27: pnres in lloxn inue ieti rts y (-) Jin, K.S., M.K. Kthirvn, R.S. Somni n C.J. epietehin. Life. Si., 13: Shishoo, The iology n hemistry of Chue, M.A., O.E. Oriskwe, O.J. Afonne, K.S. hyperlipiemi, Bioorg. Me. Chem., 15: Gmenil, O.H. Vongtu n E. Oi, Jykr, B. n B. Suresh, Antihyperglyemi n Hypoglyemi effet of the queous extrt of hypoglyemi effet of Aporos linleyn in Boerrhvi iffus leves. In. J. Phrmol., 33: lloxn-inue ieti rts. J. Ethnophrmol., : Cohn, R.M. n K.S. Roth, Biohemistry n Jyweer, D.M.A., Meiinl plnts use in isese. Willims n Klikins Pulishers, Ceylon prt 2. Ntionl Siene Counil of Srilnk. Bttimore, Lipi n Lipoprotein metolism, pp: Colomio Jeen, K.J. n R. Kuttn, 1995.Antioxint tivity of De, S., B. Rvishnkr n G.C. Bhvsr, Plnts Emli offiinlis. J. Clin. Biohem. Nutr., 19: 63- with heptoprotetivetivity review. In. Drugs., 30: Kmeshwr Ro, R. Giri, M.M. Kesvulu n C. Ellmn, G.L., Tissue sulfhyryl groups. Arh. Appro, Herl meiine: In the Biohem. Biophys., 82: mngement of ietes mellitus. Mnphr Flholt, K., B. Lun n W. Flholt, An esy Vihy Ptri. I, pp: olorimetri metho for routine etermintion of free Kirkosyn, A., E. Seymour, O.B. Kufmn, S. Wrer, S. ftty is in the plsm. Clin. Chim. At., 46: 105- Bolling n S.C. Chng n Antioxint 111. pity of polyphenoli extrts from leves of Folh, J., M. Lees n S.G.H. Solne, A simple Crtegus levigt Crtegus monogyn metho for isoltion n purifition of totl lipis (Hwthorn) sujete to rought n ol stress. J. from niml tissues. J. Biol. Chem., 26: Agri. Foo Chem., 51: Grry, R. n Buethner, The peking orer of free Krishnkumr, K., K.T. Augusti n P.L. Vijymml, rils n ntioxints: Lipiperoxition, " Hypolipiemi effet of Sli olong wll toopherol n sorte. Arh. Biohem. Biophys., root rk in STZ ieti rts. Me. Si. Res., 28: 300: Goomn, L.S. n A. Gilmn, The Kweiterovih, P.O., Metoli pthwy of high phrmologil sis of therpeutis, 7th En., MMilln, New York, pp: 490. Gulti, R.K., S. Agrwl n S.S. Agrwl, Heptoprotetive stuies on Phyllnthus emli L. n queretin. In. J. Exp. Biol., 33: Gruny, S.M., Comprison of monounsturte ftty i n rohyrtes for lowering plsm holesterol. N. Eng. J. Me., 314: Hkim, Z.S., B.K. Ptel n R.K. Goyl, Effet of hroni rmipril tretment in streptozotoin-inue ieti rts. In. J. Physiol. Phrmol., 41: NIH Consens Sttement Helth implitions of oesity, 5: 1-7. Holmn, R.R. n R.C. Turner, Orl Agents n Insulin in the Tretment of Dietes, Blkwell, Oxfor, 1991, pp: Hu, F.B., J.E. Mnson n W.C. Willett, Types of ietry ft n risk of oronry hert isese: A ritil review. J. Am. Coll. Nutr., 19: 5. Inofers, H. n H. Sies, The proution y sorte n glutthione ginst mirosoml lipi lipi peroxition is epenent on vitmin E. Eur. J. Biohem., 174: ensity lipoprotein n triylglyerols. Criol., 86: MGown, M.W., J.D. Artiss, D.R. Strnergh n B. Zk, A peroxise-ouple metho for the olorimetri etermintion of serum triglyeries. Clin. Chem., 29: Mishr, M., U.N. Pthk n A.B. Khn, Emli offiinlis Gertn n serum holesterol level in experimentl rits. Br. J. Exp. Pthol., 62: Momin, A., Role of inigenous meiine in primry helth re, in proeeings of First Interntionl Seminr on Unni Meiine. New Delhi, Ini, pp: 54. Moreno, J.J. n M.T. Mitjvli, The egree of unsturtion of ietry ftty is n the evelopment of theroselerosis. J. Nutr. Biohem., 14: 182. Oetol, A.A., O. Akinloye, C. Egujoi, W.A. Aenkunle n A.O. Ayool, Possile ntiieti n ntihyperlipiemi effet of fermente Prki Bigloos (JACQ) extrt in lloxn-inue ieti rts. Clin. Exp. Phrmol. Physiol., 33:

9 Pk. J. Nutr., 9 (1): 43-51, 2010 Ppio, G., F.A. Pisnthi, M.Y. Ltrronio, E. Ammenol n M. Glieri, Multiple lowose n single high ose tretments with streptozotoin o not generte nitri oxie. J. Cell. Biohem., 77: Prksh, J., S.K. Gupt n N. Singh, Chemopreventive tivity of Withni Somnifer in experimentlly inue firo srom tumors in Swiss lino rts. Phytother Res., 15: Prine, P.S.M. n V.P. Menon, Antiieti tivity of Tinospor orifoli roots in experimentl ietes. J. Ethnophrmol., 65: Pushprj, P., C.H. Tn n B.K.H. Tn, Effets of Averrho ilimi lef extt on loo gluose n lipis in streptozotoin-ieti rts. J. Ethnophrmol., 72: Rjsekrn, S., K. Rvi, K. Sivgnnm n S. Surmnin, Benefiil effets of loe ver lef gel extrt on lipi profile sttus in rts with STZ-inue ietes. Clin. Exp. Phrmol. Physiol., 33: Rjkumr, L., N. Srinivsn, K. Blsurmnin n P. Govinrjulu, Inrese egrtion of erml ollgen in ieti rts. In. J. Exp. Biol., 29: Roe, J.H. n C.A. Kuether, The etermintion of sori i in whole loo n urine through the 2,4-initrophenylhyrzine erivtive of ehyrosori i. J. Biol. Chem., 11: Su, M.C. n R. Kuttn, Anti-ieti tivity of meiinl plnts n its reltionship with their ntioxint property. J. Ethnophrmol., 81: Sski, T., S. Mtsy n A. Sore, Effet of eti i onentrtion on the olour retion in the O- toluiine ori i metho for loo gluose estimtion. Rinsho Kgrku, 1: Shefer, E.J., Lipoproteins, nutrition n hert isese. Am. J. Clin. Nutr., 75: 191. Shrm, S.B., A. Nsir, K.M. Prhu, P.S. Murthy n G. Dev, Hypoglyemi n hypolipiemi effet of ethnoli extrt of sees of Eugeni jmoln in lloxn-inue ieti rits. J. Ethnophrmol., 85: Shirwkr, A., K. Rjenrn, C.D. Kumn n R. Bol, Antiieti tivity of queous lef extrts of Annon squmos in STZ-inue niotinmie type 2 ieti rts. J. Ethnophrmol., 91: Sim, A.A. n K. Sugimoto, Experimentl ieti neuropthy: An upte. Dietologi., 42: Su, V., T. Murugesn, R. Bhskr Ro, L. Ghosh, M. Pl, S.C. Mnl n B.P. Sh, Antiieti potentilof Brleri lupulin extrt in rts. Fitoterpi., 75: 14. Suresh Kumr, J.S. n V.P. Menon, Peroxitive hnges in experimentl ietes mellitus. In. J. Me. Res., 96: Tskinen, M.R., Lipoprotein lipse in ietes. Di. Met. Rev., 3: Thkur, C.P., B. Thkur, B. Singh, S. Singh, P.K. Sinh P.K. n S. K. Sinh, The rits. Ayurvei meiines, Hritki, Aml n Bhir reue holesterol inue theroslerosis in In. J. Criol., 21: Tunli, S. n R. Ynrg, Effet of Vnyl sulfte on the sttus of lipi prmeters n on stomh n spleen tissues of Stz-inue ietes. Phrmol. Res., 53: Twij, H.A.A. n A.A. Al-Br, Hypoglyemi tivity of Artemisi her l. J. Ethnophrmol., 24: Vlithn, M.S., Heling plnts. Curr. Si., 75: Worl Helth Orgnistion (WHO), Seon Report of the WHO Expert Committee on Dietes Mellitus. Tehnil Report Series, 646: 66. Zvroni, I., E. Bonr n M. Pgilor, Risk ftors for oronry rtery isese in helthy persons with hyperinsulinemi n norml gluose tolerne. New. Eng. J. Me., 320: Zilversmit, D.B. n A.K. Dvis, Miro etermintion of plsm phospholipis y trihloroeti i preipittion. J. L. Clin. Me., 35: