Pharmacodynamic Interaction of Fenugreek Seeds Extract with Fluvastatin in Diabetes Induced Vascular Dysfunction

Transcription

1 ISSN Aville Online t Interntionl Journl of Phrmeutil & Biologil Arhives 2012; 3(4): ORIGINAL RESEARCH ARTICLE Phrmodynmi Intertion of Fenugreek Seeds Extrt wi Fluvsttin in Dietes Indued Vsulr Dysfuntion Preml Kpdi N*, A K Prsd, Shnkul kumr, Stish V Iyer, Hitesh A Ptel, Jtin A Ptel G.H.B. College of Phrmy, Aniyd, Gujrt, Indi Reeived 12 My 2012; Revised 09 Aug 2012; Aepted 18 Aug 2012 ABSTRACT This study is minly imed to investigte e erpeuti potentil of FG seeds in omintion wi e well-known hypolipidemi t is fluvsttin nd hypoglyemi drugs pioglitzone nd glipizide for e tretment of vsulr dysfuntion indued y dietes mellitus. Clinil studies hve lredy proven t fluvsttin erpy n signifintly redue e rdiovsulr moridity nd mortlity euse of its lipid- nd holesterol-lowering potentil. The long-term effiy of pioglitzone nd glipizide s single nd omined erpy ginst e dietes mellitus hs een explored widely. In ddition to eir gluose-lowering effets, pioglitzones n modulte numer of rdiovsulr risk ftors vi eir Anti-inflmmtory, nti-erogeni nd nti-romoti properties. Similrly, e erpy of glipizide is found to e highly potent for e tretment of dietes mellitus, s e drug lowers lood gluose level very signifintly. Alough e omined erpy of stndrd drugs is found to e highly effetive in normlizing e norml iohemil prmeters in dieti ptients, e FG seed extrt lone or in omintion wi hypoglyemi drugs, pioglitzone nd glipizide is lso oserved to e very promising erpy for dietes mellitus. The FG seed extrt in omintion wi pioglitzone nd glipizide shows e gluose-lowering effet. Erlier Fluvsttin ws proved for normlizing endoelil dependent relxtion. Even e endoelil dependent relxtion ws improved in treted groups. Thus, eir omintion is ssumed to e very potentil tretment for vsulr dysfuntion indued y dietes mellitus. Key words: Hypolipidemi, Hypoglyemi, vsulr dysfuntion, Fenugreek seed (FG). INTRODUCTION Endoelil dysfuntion n e defined s e prtil or omplete loss of lne etween vsoonstritors nd vsodiltors, grow promoting nd inhiiting ftors, proerogeni nd nti-erogeni ftors, nd pro-ogulnt nd ntiogulnt ftors. Endoelil dysfuntion now regrded s n erly pivotl event in erogenesis [1].Endoelil dysfuntion in e rhil rtery highly orreltes wi endoelil dysfuntion in e oronry irultion, whih is emerging s n independent risk ftor for vsodiltion, respetively) rdiovsulr disese [2].There is strong ssoition etween endoelil dysfuntion nd oxidtive stress nd development of oer mirovsulr omplitions [3].Nitri oxide is importnt sustne sereted from endoelium so lled endoelium-derived relxing ftor [4]. Nitri oxide not only produes vsodiltion, ut it lso prtiiptes in vrious proesses t re enefiil to e vsulture, suh s e redution of vsulr smoo musle ell migrtion nd grow, pltelet ggregtion nd romosis, monoyte nd mrophge dhesion, nd inflmmtion. There re vrious phrmologil nd non-phrmologil stimuli t n e used to ssess e prodution of nitri oxide in endoelil ells in vivo s well s its effet on e underlying vsulr smoo musle ells (endoelium-dependent nd -independent [5].The min mehnism underlying endoelil dysfuntion is oxidtive stress oer mehnism is prodution of AGEs nd on turn produing Retive oxygen speies [6].nd vsulr endoelil dysfuntion is leding use for peripherl rtery diseses [7].so ssoition of o dietes nd endoelil vsulr dysfuntion my led ptient to de. Mny ommeril drugs ville whih n modify e disese pttern. Among ese drugs, ere re two lsses t re widely used in *Corresponding Auor: Shnkul Kumr, Emil: Kumr.snkul@gmil.om, Phone No:

2 Shnkul Kumr et l. / Phrmodynmi Intertion of Fenugreek Seeds Extrt wi Fluvsttin in Dietes Indued Vsulr Dysfuntion people wi oesity nd type 2 dietes. These re e izolidenediones (TZDs) nd e igunides (metformin). TZDs diretly redue insulin resistne y enhning insulin tion in peripherl tissues nd erefore, hve een ttrtive ndidtes for studying e effets on vsulr funtion of reduing insulin resistne. Severl studies showed e signifint impt of troglitzone on glyemi, insulin resistne, nd trditionl rdiovsulr risk ftors. There re two urrently ville TZDs, pioglitzone nd rosiglitzone. They o signifintly redue insulin resistne nd improve glyemi ontrol in people wi type 2 dietes. They lso seem to hve n overll positive effet on lood pressure, lipids, nd ft distriution [8,9]. A good omintion erpy n e used to get strit glyemi ontrol. Fluvsttin tretment for dyslipidemi in ptients wi type 2 dietes mellitus on insulin erpy is effetive nd well tolerted [10]. The hypoglyemi effet of fenugreek seeds is studied previously [11]. The phrmodynmi intertion of Fenugreek seed extrt (Rih in polyphenol) nd fluvsttin in dieti indued vsulr dysfuntion is not studied until now. Hene n ttempt is mde to study e effet of Fenugreek seed extrt wi fluvsttin in dietes indued vsulr dysfuntion in rts. MATERIALS AND METHODS Colletion of plnt mterils: Fenugreek seed ws olleted from Shool of siene, Gujrt University, Ahmdd. It ws identified nd uentited y Dr. Himnshu A. Pndy, Assoite Professor, Shool of siene, Gujrt University, Ahmdd. Preprtion of extrt: Fenugreek seeds (100 g) were finely powdered, mixed wi 80% menol nd kept t room temperture for 5 dys. After 5 dys it ws filtered nd e solvent ws evported. The residue ws dissolved in wter nd e queous lyer ws wshed wi petroleum eer severl times until ler upper lyer of petroleum eer ws otined. The lower lyer ws en treted wi eyl ette ontining glil eti id (10 ml/l).extrtion of polyphenols ws rried out for 36 h t room temperture nd e omined eyl ette lyer ws onentrted). The residue ws lyophilized nd stored. This yielded out 6 8 g per 100 g of seed powder. A Menoli extrt ws prepred nd used for studies. Experimentl nimls: Mle Sprgue dwley Rts weighing g were housed t 25 ± 5 C in well ventilted Animl house under 12:12 h light drk yle. The nimls were mintined under stndrd onditions in n niml house s per e guidelines of Committee for e Purpose of Control nd Supervision on Experiments on Animls1359//10/ (CPCSEA). INDUCTION OF DIABETES: The nimls were fed wi HFD one dy for two weeks followed y I.P injetion of streptozotoin (35mg/kg) dissolved in 0.5M/l itrte uffer ( ph: 4.4) fter overnight fsting. STZ injeted nimls were en given 5% w/v gluose solution for 5-6 hours following e injetion to prevent initil drug indued hypoglyemi mortlity. The rts wi non fsted plsm level 300mg/dl were onsidered dieti. The lood Smple ws olleted from til vein nd lood gluose ws heked using gluose dignosti kit. STANDARDISATION AND SELECTION OF LOW DOSE FOR STANDARD DRUGS: Orl Gluose Tolerne Test: After overnight fsting (18hrs), 0-min lood smple ws tken from e tip of e til of eh rt of different groups. Gluose solution (2 g/kg P.O) ws given fter 30 min fter e dministrtion of e drug. Four more smples were tken t 30, 60, 90 nd 120 min fter gluose dministrtion. All lood smples were heked wi e help of uhek gluometer. The orl gluose tolerne test will e performed on overnight fsted Sprgue dwley rts. Rts will e divided into different groups s follows: Group 1: Norml ontrol, rts reeive sline/vehile. Group 2: Dieti ontrol. Group 3: Fenugreek seeds extrt (200 mg/kg) Group 4: Pioglitzone (10 mg/kg) Group 5: Glipizide (5 mg/kg) Effet of fenugreek seed extrt lone nd in omintion wi orl hypoglyemi gents in streptozotoin indued dieti vsulr dysfuntion [12,13,14] : Rts ws divided into different groups (n=6) s follows nd e tretment wi e respetive drugs ws given for 8 weeks. The estimtions were rried out on e 15, 30, 45 nd 60 dys of e tretment. Group 1: Norml ontrol, rts reeive sline/vehile. 901

3 Shnkul Kumr et l. / Phrmodynmi Intertion of Fenugreek Seeds Extrt wi Fluvsttin in Dietes Indued Vsulr Dysfuntion Group 2: Dieti ontrol Group3: FSEt (200 mg/kg p.o) Group 4: Pioglitzone (10 mg/kg) Group 5: Glipizide (5mg/kg) Group 6: Pioglitzone (7.5mg/kg) plus Glipizide (3.75mg/kg) Group 7: Pioglitzone(7.5 mg/kg )+ FSEt(200mg/kg) Group 8: Glipizide (3.75 mg/kg) + FSEt (200 mg/kg) STATISTICAL ANALYSIS: Dt re presented s e men ± SE from 6 rts per group. Comprison of men vlues mong e vrious groups ws performed y one wy ANOVA. For e single omprison etween e groups unpired Student s t-test ws used. RESULTS AND DISSCUSSION The phrmodynmi intertion of fenugreek seed extrt (Rih in polyphenol) nd fluvsttin in dieti indued vsulr dysfuntion is not studied until now. Hene n ttempt is mde to study e effet of fenugreek seed extrt wi fluvsttin in dietes indued vsulr dysfuntion in rts nd result is tulted in tle. The present study (OGTT) lso indites profound effet of FSEt (Fenugreek seed extrt). FSEt showed signifint inrese in gluose tolerne when ompred wi dieti ontrol, whih might e due to insulinotropi property in vitro nd improved gluose tolerne in rt model of type 2 DM1 [15]. In type 2 dieti ptients, e ingestion of 15 g of powdered (wter soked) fenugreek seed signifintly redued postprndil gluose levels during e gluose tolerne test [16]. The omined erpy of FSEt wi glipizide showed more signifine in reduing e Tle 1: Effet of FSEt nd its omintion wi Orl Hypoglyemi Agents on OGTT in Norml rts TIME FBS 0 min 30 min 60 min 120 min NC 84± ± ±0.8 ( 16%) ±1.5 ( 10%) 86.2± 0.6 ( 17%) DC 205± ± ±2.6 ( 35%) ±3.0 ( 34.5%) 285.4±2.1 ( 33%) FSEt 86.3 ± ± ± 4.1 ( 18.68%) ±1.9 ( 14%) 88.90±0.8 ( 4%) P 85.6 ± ± ±3.4 ( 29%) 11( 20%) 2.33± ±2.8 ( 3%) G ± ± ±3.1 ( 28.8%) ±0.6 ( 13%) 84.99±0.5 ( 6%) P+G ± ± ±0.5 ( 17.7%) 99 ±0.8 ( 9%) 70.22± 0.9 ( 23%) P+ FSEt 86.1 ± ± ±0.9 ( 20%) 102 ±1.8( 15%) 85.77± 0.8 ( 5.1% ) G+ FSEt 82.66± ± ±3.8 ( 15.9%) 96± 2.2 ( 11.6%) 82.1± 0.7 ( 5%) All vlues re men ± SEM, n=6, P<0.001, P<0.01, P<0.05 when ompred to norml ontrol group. Tle 2: Effet of FSEt nd its omintion wi Orl Hypoglyemi Agents on OGTT in Dieti rts TIME FBS 0 min 30 min 60 min 120 min NC ± ± ±2.0 ( 6%) ± 0.9 ( 15.3%) 90.21± 2.1 ( 2.8%) DC ± ± ± 0.8 ( 17.5%) ±0.8 ( 40%) 306.3±0.1 ( 44%) FSEt ± ± ± 3.3 ( 10.4%) 222 ±1.2 ( 17%) 186.1±1.7 ( 2%) P ± ± ±3.2 ( 17.4%) ±4.1( 27%) 212.7±5.0 ( 5.5%) G ± ± ±2.4 ( 12.56%) 243.5±4.3( 20%) 180±2.9 ( 12.4%) P+G ± ± ±0.8 ( 8%) 208±1.9 ( 19%) 143.1±1.8( 18%) P+ FSEt ± ±3.9 ( 11%) ±4.0 ( 11%) ±2.8 ( 30%) 194.8±1.8 ( 14%) G+ FSEt ± ± ±2.2 ( 3%) ±2.1 ( 15%) 187.4±0.6 ( 2%) All vlues re men ± SEM, n=6, P<0.001, P<0.01, P<0.05 when ompred to dieti ontrol group. postprndil gluose levels when ompred wi individul erpy. When e dieti rts were treted for 10 weeks, e omintion erpy (FSEt wi glipizide) did not produe hypoglyemi effet ut e omined erpy of (Orl hypoglyemi gents) OHA s hd produed hypoglyemi. The prodution of NO from e endoelil ells is stimulted y insulin [17]. Thus, in hely non-dieti sujets, insulin inreses endoelium-dependent (NO-medited) vsodilttion ut endoelium- dependent vsodilttion is redued in insulin-resistnt sujets [18]. Insulin-medited gluose disposl is orrelted inversely wi e severity of e impirment in endoelium-dependent vsodiltion [19]. Drug erpies whih inreses insulin sensitivity (metformin nd izolidinediones), improves endoeliumdependent vsodilttion [20,21]. Hyperglyemi indues series of ellulr events t inrese e retive oxygen speies (ROS) prodution (suh s superoxide nion) t intivte NO (Nitri Oxide) to form peroxynitrite [22,23]. Dieti rt endoelium produes n enhned rte of superoxide nion genertion [24]. It is importnt to undersore e ft t e superoxide is n initil oxygen rdil speies whih my led to seondry rdils or retive oxygen speies suh s H 2 O 2 or hydroxyl rdils [25]. The present study (FSEt wi low dose of OHA s) indites e signifint improvement in endoelil dependent vsodilttion euse e perentge relxtion produed y etylholine ws more signifint when ese groups were ompred wi dieti ontrol. All e results re tulted in (Tle 1, 2, 3 & 4). 902

4 Shnkul Kumr et l. / Phrmodynmi Intertion of Fenugreek Seeds Extrt wi Fluvsttin in Dietes Indued Vsulr Dysfuntion Tle 3: Effet of FSEt nd its omintion wi Orl Hypoglyemis on Blood Gluose (mg/dl) Groups 0 dy 15 dy 30 dy 45 dy 60 dy 75 dy NC 124± ± ± ± ± ±0.8 DC 265± ± ± ± ± ±0.4 FSEt 241± ± ±0.5 P 245± ± ± ± ± ± ±0.6 G 270± ± ± ± ±0.4 P+G 241± ± ± ± ±0.5 P+FSEt 251± ± ± ± ±1.5 G+FSEt 262± ± ± ± ±0.8 All vlues re men ± SEM, n=6, P<0.001, P<0.01, P<0.05 when ompred to dieti ontrol group 139± ± ± ± ±0.1 Tle 4: Comprison of relxnt responses in treted groups: TREATMENT Perentge relxnt response to Aetylholine SNP 0.1 (µm) 0.3 (µm) (1.0 µm) NORMAL (Men ±SEM) ± ± ± ±1.98 DIABETIC (Men ±SEM) 9.07 ± ± ± ±2.87 FSEt (Men ±SEM) ± ± ± ±3.12 PIO (Men ±SEM) ± ± ± ±1.87 GLIP (Men ±SEM) ± ± ± ±2.34 PIO+GLIP (Men ±SEM) ± ± ± ±3.05 FSEt+PIO (Men ±SEM) ± ± ± ±1.05 FSEt+GLIP (Men ±SEM) ± ± ± ±3.76 FSEt+FLU (Men ±SEM) ± ± ± ±1.89 P+FLU (Men ±SEM) ± ± ± ±2.67 G+FLU (Men ±SEM) ± ± ± ±2.23 All vlues re men ± SEM, n=6, P<0.001, P<0.01, P<0.05 when ompred to dieti ontrol group. CONCLUSION Endoelil dysfuntion ssoited wi insulin resistne ppers to preede e development of over hyperglyemi in ptients wi type 2 dietes mellitus. Therefore, endoelil dysfuntion my e ritil erly trget for e prevention of eroslerosis nd CVD in ptients wi dietes mellitus or insulin resistne. Therefore preventive erpy wi omintion of FSEt wi OHA s (low dose) my prevent e endoelil dysfuntion in dieti ptients (predieti stte) due to free rdil svenging tivity, metoli ontrol. ACKNOWLEDGEMENT The uors wish to nk Chirmn, Exeutive Diretor, Goplk viks mndl; GHB College of Phrmy, Aniyd for eir filities provided for is study. REFRENCES 1. Enrique CA. Endoelil dysfuntion in oesity nd insulin resistne: rod to dietes nd hert disese. Oes Res 2003; 11: Shhinger V, Britten MB, Zeier AM. Prognosti impt of oronry vsodiltor dysfuntion on dverse long-term outome of oronry hert disese. Cirultion 2000; 101: Son SM. Role of vsulr retive oxygen speies in development of vsulr normlities in dietes. Dietes Res Clin Pr 2007; 77: Cokroft J R. Exploring Vsulr Benefits of Endoelium-Derived Nitri Oxide. A J H 2005; 18: Cllero E. Endoelil Dysfuntion in Oesity nd Insulin Resistne: A Rod to Dietes nd Hert Disese. Oes Res 2003; 11: Esper RJ, Nordy RA,Vilrino JO, Prgno A, Chrron JL, Mhdo AR. Endoelil dysfuntion: omprehensive pprisl. Crdiovs Dietol 2006; 5: Brevetti G, Shino V, Chiriello M. Endoelil dysfuntion: A key to e pophysiology nd nturl history of peripherl rteril disese? Aeroslerosis 2008; 197: Kenned DL, Piper J, Bun C. Trends in use of orl hypoglyemi gents Dietes Cre 1988; 11: Prulkr AA, Pendergrss ML, Grnd- Ayl R, Lee TR, Fonse VA. Nonhypoglyemi effets of izolidinediones. Ann Intern Med 2001; 134:

5 Shnkul Kumr et l. / Phrmodynmi Intertion of Fenugreek Seeds Extrt wi Fluvsttin in Dietes Indued Vsulr Dysfuntion 10. Visseren FLJ, Boutter PK, Vn Loon BJP, Erkelens WD. Tretment of Dyslipidemi wi Fluvsttin in Ptients wi Type 2 Dietes Mellitus: Effets on Lipids, Mentl Stte nd Firinolysis. Clin Drug Invest 2001; 21: Aryne MA, Sultn N, Mirz AZ, Zueri MH, Siddiqui FA. In vitro hypoglyemi tivity of Menoli extrt of some indigenous plnts. Pk J Phrm Si 2007; 20: Xi J, Allenrnd B, Sun GY. Dietry supplementtion of grpe polyphenols nd hroni enol dministrtion on LDL oxidtion nd pltelet funtion in rts. Life Si 1998; 63: Lee MY, Lee EY, Lee BJ, Won CS, Koh JH. Benefiil effet of izolidinediones on dieti neuropy in OLETE rts. Yom sei Med J 2007;48: Hosszufusi N, Reinhers L, Tkei S, Chn E, Chrles MA. Glipizide indued prevention of dieti nd utoimmune events in BB rts. J A Autoimmun 1994; 9: Rju J, Gupt D, Ro AR, Ydv PK, Bquer NZ.Trigonell foenum greum (fenugreek) seed powder improves gluose homeostsis in lloxn dieti rt tissues y reversing e ltered glyolyti, gluoneogeni nd lipogeni enzymes. Mol Cell Biohem.2001; 224: Mohmmd S, Th A, Bmezi RN, Bquer NZ. Modultion of gluose trnsporter (GLUT4) y vndte nd Trigonell in lloxn-dieti rts. Life Si; 2006; 78(8): Kuoki K, Jing ZY, Tkhr N. Regultion of endoelil onstitutive nitri oxide synse gene expression in endoelil ells nd in vivo: speifi vsulr tion of insulin. Cirultion 2000; 101: Lkso M, Edelmn SV, Brehtel G, Bron AD. Deresed effet of insulin to stimulte skeletl musle lood flow in oese mn: novel mehnism for insulin resistne. J Clin Invest 1990; 85: Mer K, Lkso M, Edelmn S. Evidene for physiologil oupling of insulin-medited gluose metolism nd lim lood flow. Am J Physiol Endorinol Met 2000; 279: Mer KJ, Verm S, Anderson TJ. Improved endoelil funtion wi metformin in type 2 dietes mellitus. J Am Coll Crdiol 2001; 37: Wtne Y, Sunym S, Shimd K, Swno M,Hoshi S,Iwn Y. Troglitzone improves endoelil dysfuntion in ptients wi insulin resistne. J Aerosler Throm 2000; 7: Bekmn JA, Goldfine AB, Gordon MB. Asorte restores endoeliumdependent vsodiltion impired y ute hyperglyemi in humns. Cirultion; 2001; 103: Nishikw T, Edelstein D, Du XL Normlizing mitohondril superoxide prodution loks ree pwys of hyperglyemi dmge. Nture.2000; 404: Pieper GM. Oxidtive stress in dieti lood vessels. FASEB J 1995;9:A Ohkuw T, Sto Y, Noi M. Hydroxyl rdil formtion in dieti rts indued y streptozotoin. Life Si 1995;56: