Antihypertensive and Antioxidant Potential of Borneol-A Natural Terpene in L- NAME Induced Hypertensive Rats

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1 ISSN Aville Online t Interntionl Journl of Phrmeutil & Biologil Arhives 2010; 1(3): ORIGINAL RESEARCH ARTICLE Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L- NAME Indued Hypertensive Rts Murugesn Srvn Kumr, Surmniyn Kumr, Booln Rj * *Deprtment of Biohemistry nd Biotehnology, Annmli University, Tmil Ndu, Indi. Reeived 12 My 2010; Aepted 7 July 2010 ABSTRACT The present study ws imed to investigte the ntihypertensive nd ntioxidnt potentil of orneol - nturl terpene, ginst N ω -Nitro-L-rginine methyl ester hydrohloride (L-NAME) indued hypertension in rts. Hypertension ws indued in dult mle lino rts of the Wistr strin, weighing g, y orl dministrtion of the L-NAME (40 mg/kg w/dy) in drinking wter for 4 weeks. Rts were treted with orneol (50 mg/kg w/dy) for 4 weeks. Systoli nd distoli lood pressure ws mesured every week nd the toxi effet of L-NAME ws determined using lipid peroxidtive mrkers (thiorituri id retive sustnes (TBARS) nd lipid hydroperoxides (LOOH). We ssessed the tivity of enzymti ntioxidnts (superoxide dismutse (SOD), tlse (CAT), glutthione peroxidse (GPx) nd mesured the levels of non-enzymti ntioxidnts (vitmin C, vitmin E nd redued glutthione (GSH)) levels in erythroytes, plsm nd tissues. Our results showed tht orl dministrtion of orneol (50 mg/kg w/dy) signifintly ttenuted systoli nd distoli lood pressure. In ddition, orneol signifintly redued lipid peroxidtion nd inresed the tivities nd level of enzymti nd nonenzymti ntioxidnts. These results were supported y histopthlogil studies. The effet of orneol ws omprle with nifedipine. These findings suggest tht orneol ffords signifint ntihypertensive nd ntioxidnt effet ginst L-NAME indued hypertensive rts. Keywords: Hypertension, Borneol, Lipid peroxidtion, Antioxidnts, L-NAME. INTRODUCTION Crdiovsulr diseses (CVD) re the leding use of deth worldwide, ounting for n estimted 14 million deths in 1990 nd projeted to use 25 million deths in 2020 [1]. Hypertension is onsidered to e mjor risk ftor in the development of CVD [2]. Therefore, it hs een reeived inresing ttention y reserhers. Currently ville ntihypertensive drugs ply n importnt role in the tretment of hypertension. However, ommonly used ntihypertensive drugs re expensive with mny dverse effets. Therefore there is need for popultion-sed, ost-effetive, dverse-effet free hypertension ontrol strtegies to e developed [3]. The development of sfe nd effetive wy to mnge hypertension hs hllenged medil reserhers for enturies. In reent times, fous on plnt reserh hs inresed ll over the world nd lrge ody of evidene ws olleted to show immense potentil of mediinl plnts used in vrious trditionl systems. A wide vriety of the trditionl herl remedies re used y hypertensive ptients, espeilly in the third world ountries nd my therefore represent new venues in the serh for lterntive ntihypertensive drugs [4]. Monoterpenes re primry ompounds of plnt essentil oils nd the effets of mny mediinl hers hve een ttriuted to them.borneol, iyli monoterpene, present in the essentil oils of numerous mediinl plnts is often used in trditionl Chinese mediine nd is very importnt ingredient in mny Jpnese inense formuls with uplifting effets on the mind [5] nd on the neurotrnsmitter GABA [6]. In folk remedies, orneol is used for the tretment of dominl pin, prtiulrly stomhhe [7]. Borneol is used more frequently for topil pplitions suh s to injuries, urns, rheumti *Corresponding Author: Dr. B. Rj, Emil: drrju@gmil.om, Contt No:

2 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts pins, hemorrhoids, skin diseses nd ulertions of the mouth, er, eye, nd nose. Moreover, it stimultes digestive system y inresing prodution of gstri juies; tones the hert nd improves irultion; trets ronhitis, oughs nd olds; redues swelling; relieves stress; nd n e used s toni to promote relxtion nd redue exhustion [8]. It shows inhiitory effets on severl Grm (-) nd Grm (+) pthogeni miroorgnisms, exhiits mrked ntifungl tivity [9] nd rdil svenging properties [10]. Immunomodultory effets of orneol were studied y Juh s [11] showed tht orneol ws le to signifintly suppress the pro-inflmmtory ytokine mrna expression in oloni inflmmtion in mie. In ddition, orneol ws reported to t s iotive mteril in the ellulr signl trnsdution system [12]. Reently orneol hs een demonstrted to show ntithromoti nd ntipltlet tivity [13]. Our im, therefore, ws to exmine whether dministrtion of orneol ould hve positive effets on lood pressure nd oxidtive sttus in L-NAME indued hypertensive rts. MATERIALS AND METHODS Animls Mle lino Wistr rts, 7-8 weeks old (weighing g) were proured from the Centrl Animl House, Deprtment of Experimentl Mediine, Rjh Muthih Medil College nd Hospitl, Annmli University nd mintined in n ir-onditioned room (25 ± 1ºC) with 12 h light/12 h drk yle. Feed nd wter were provided d liitum. All the experimentl studies were onduted in the Deprtment of Biohemistry, Fulty of Siene, Annmli University, in ordne with the Ntionl Institutes of Helth Guide for the Cre nd Use of Lortory Animls (NIH 1985); the experimentl study ws pproved y the Ethil Committee of Rjh Muthih Medil College nd Hospitl (Reg No.160/1999/CPCSEA, Pro. No.597), Annmlingr, Tmil Ndu. A pilot study ws onduted with three different doses of orneol (25, 50 nd 100 mg/kg) to determine the dose dependent effet of orneol in L-NAME indued hypertensive rts. It ws oserved tht fter 14 dys of experiment, orneol tretment t the doses of 25, 50 nd 100 mg/kg signifintly (p< 0.05) lowered the elevted lood pressure in L-NAME hypertensive rts. 50 mg/kg of orneol showed higher signifint effet thn lower dose 25 mg/kg nd the higher dose 100. Hene, we hve hosen the dose (50 mg/kg) for our study. Chemils Borneol nd N ω -Nitro-L-rginine methyl ester hydrohloride (L-NAME) were purhsed from Sigm-Aldrih Compny (St. Louis, Missouri, USA). All other hemils used were of nlytil grde otined from E. Merk, Mumi nd HIMEDIA, Mumi, Indi. Experimentl design The rts were rndomly divided into five groups of six nimls eh s given elow. The first group (n=6) served s ontrol nd reeived tp wter only. In the seond group (n=6) served s drug (orneol) ontrol group reeived orneol t dose of 50 mg/kg /dy. The third group, L-NAME group (n=6), L-NAME ws dded to drinking wter t onentrtion of 0.4 mg/ml to ount for dily intke of 40 mg/kg [14].The fourth group (orneol group, n=6) reeived simultneously L- NAME (40mg/kg/dy) nd orneol t 50 mg/kg/dy nd the lst group (nifidipine group, n=6) reeived simultneously L-NAME (40 mg/kg/dy) nd nifedipine (20 mg/kg/dy). The orneol ws suspended in 0.5% Dimethyl sulphoxide (DMSO) vehile solution nd fed y intution. Before inititing experimentl protools, mesurements of seline systoli (SBP), distoli lood pressure (DBP), men rteril lood pressure (MABP) nd hert rte (HR) y non-invsive til-uff plethysmogrphy (IITC, model 31, Woodlnd Hills, CA, USA) nd initil ody weight hve een performed. The experimentl durtion ws 30 dys. On 31 st dy the rts were srified y ervil dislotion. Blood ws olleted in dry test tue nd llowed to ogulte t mient temperture for 40 min. Serum ws seprted y entrifugtion t 2000 rpm for 10 min. The lood, olleted in heprinised entrifuge tue ws entrifuged t 2000 rpm for 10 min nd the plsm ws seprted y spirtion. After the seprtion of plsm, the uffy ot, enrihed in white ells, ws removed nd the remining erythroytes were wshed three times with physiologil sline. A known volume of erythroyte ws lysed with hypotoni phosphte uffer t ph 7.4. The hemolyste ws seprted y entrifugtion t 2500 rpm for 10 min nd the superntnt ws used for the estimtion of enzymi ntioxidnts. Hert, 2010, IJPBA. All Rights Reserved. 272

3 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts liver nd kidney tissues (250 mg) were slied into piees nd homogenised in pproprite uffer in old ondition (ph 7.0) to give 20% homogente (w/v). The homogente ws entrifuged t 1000 rpm for 10 min t 0 C in refrigerted entrifuge. The superntnt ws seprted nd used for vrious iohemil estimtions. exmintion for evidene of hypertensive tissues hnges. The ross-setionl re (CSA) of hert ws evluted from photogrphs of whole tissue setions tken t 40X mgnifition nd snned, digitized nd nlyzed y omputer, using the Adoe Photoshop Imging progrm (Adoe System Inorportion). Biohemil estimtions Lipid peroxidtion ws ssessed y mesuring the levels of thiorituri id retive sustnes (TBARS) nd lipid hydroperoxides in the plsm following the proedures of Niehus nd Smuelson [15] nd Jing [16], respetively. The tivities of erythroyte SOD, CAT nd GPx were ssyed y the method of Kkkr [17], Sinh [18]] nd Rotruk [19].The levels of non-enzymti ntioxidnts suh s sori id, α-toopherol nd redued glutthione were mesured y Roe nd Kuether [20], Bker [21] nd Ellmn [22], respetively. Histologil exmintion of rdi tissues Exised hert smple were lered of lood nd immeditely fixed in neutrl uffered solution of 10% formlin for 24 h. 5µm-thik tissues setion from hert of eh niml were prepred from proessed prffin-emedded smples. Setion were stined with Hemtoxylin nd Eosin (H&E) for light mirosopi Effet of orneol on ody weight, tissue, wter intke in norml, L-NAME indued hypertensive rts Tle 1 shows the ody weight, tissue weights nd wter intke of ll groups. The initil ody weights (BW) were 181 ± 1.16, 183 ± 1.64, 185 ± 1.47, 184 ± 1.64,183 ± 2.42g (dt not shown), nd finl odyweights were 206 ± 1.75, 208 ± 1.47, 160 ± 3.61, 204 ± 1.94, 208 ± 3.25 for ontrol, ontrol + orneol, L-NAME (40mg/kg/dy), L-NAME + orneol (50mg/kg/dy), L- NAME + nifedipine (20 mg/kg/dy) groups, respetively. L-NAME dministered Sttistil nlysis Dt were nlysed y one wy nlysis of vrine (ANOVA) followed y Dunn s multiple rnge test (DMRT) using sttistil softwre pkge (SPSS for Windows, V. 11.5, Chigo, USA). Results were presented s mens ± S.D. p-vlues < 0.05 were onsidered s sttistilly signifint. RESULTS Blood pressure mesurements As shown in fig. 1 nd 2 dministrtion of L-NAME in drinking wter for 4 weeks signifintly elevted SBP (182.3 ± 4.07 mm Hg), DBP (118.8 ± 1.69 mm Hg), s ompred to tht of ontrol rts (SBP (112.3±2.74 mm Hg), DBP (72.2±2.41 mm Hg, p< 0.05). Tretments with orneol nd nifedipine signifintly ttenuted development of high lood pressure (SBP (120.2±3.41mm Hg), DBP (74.65±1.69 mm Hg) nd nifedipine (SBP (116.3±5.81 mm Hg), DBP (72.5±3.27 mm Hg p< 0.05). rts hd signifintly inresed wter intke, kidney nd hert weight. Borneol nd nifedipine signifintly redued the wter intke, renl nd hert weight in L-NAME rts. The BWs of rts in the five groups differ signifintly t the eginning or t the end of study. Tle.1 Effet of orneol on ody weight, reltive tissue weights nd wter intke in ontrol nd L-NAME-indued hypertensive rts Reltive orgn weight g/100 g BW Wter intke (ml) Groups BW (g) Kidney Hert Control 206± ± ± ±1.43 Control+ orneol 208 ± ± ± ±0.98 (50 mg/kg /dy) L-NAME 160± ± ± ±1.65 (40mg/kg/dy) L-NAME + orneol 204± ± ± ±0.95 (50 mg/kg/dy) L-NAME + nifedipine (20mg/kg/dy) 208± ± ± ±2.02 Vlues re mens ± SD for six rts Vlues not shring ommon supersript differ signifintly t p < 0.05 (DMRT) 2010, IJPBA. All Rights Reserved. 273

4 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts Effet of orneol on lipid peroxidtion Tle 2 shows the onentrtion of TBARS nd lipid hydroperoxides (mrkers of lipid peroxidtion) in the plsm nd tissues (hert, liver nd kidney) of ontrol nd L - NAME treted rts. The levels of TBARS nd hydroperoxides were signifintly inresed in L- NAME rts. Orl dministrtion of orneol nd nifedipine signifintly prevented the inrese in lipid peroxidtion mrker level whih ws rought to ner ontrol. Tle 2. Effet of orneol on TBARS nd lipid hydroperoxide in the plsm nd tissues of norml nd L-NAME indued hypertensive rts. Prmeter Smples Groups TBARS LOOH Control Control + orneol (50 mg/kg BW.) L-NAME (40mg/kg BW.) L-NAME+ orneol (50mg/kg BW.) L-NAME+ nifedepine (20mg/kg BW.) Plsm (mmoles / 9.10± ± ± ± ±0.95 d dl) Liver(mmoles/ ± ± ± ± ±2.98 d g wet Kidney(mmoles/ ± ± ± ± ±9.22 d 0g wet Hert(mmoles/100 g wet 65.47± ± ± ± ±8.06 d Plsm (mg/dl) 0.13± ± ± ± ±0.02 d Liver(mmoles/ ± ± ± ± ±0.11 d g wet Kidney(mmoles 1.40± ± ± ± ±0.11 d /100g wet Hert(mmoles/ ± ± ± ± ±0.18d d g wet Vlues re mens ± SD for six rts Vlues not shring ommon supersript differ signifintly t p < 0.05 (DMRT) Enzymti ntioxidnts Tles 3 show the tivities of SOD, CAT, nd GPx in erythroyte, liver, kidney nd hert of ontrol, L-NAME nd orneol treted hypertensive rts, respetively. The tivities of these enzymti ntioxidnts were signifintly restored in erythroyte nd tissues on tretment with orneol nd nifedipine in L-NAME indued hypertensive rts. Non-enzymti ntioxidnts Tles 4 show the levels of non-enzymti ntioxidnts (vitmin C, vitmin E nd GSH) in the plsm nd tissues (liver, kidney nd hert) of ontrol,l-name nd orneol treted hypertensive rts. Our results show tht the levels of nonenzymti ntioxidnts were signifintly deresed in L-NAME indued hypertensive rts. Orl dministrtion of orneol nd nifedipine rought k these prmeters to ner ontrol. Fig 1. Effet of orneol on the systoli lood pressure (SBP) in ontrol nd L-NAME - indued hypertensive rts. Fig 2. Effet of orneol on the distoli lood pressure (DBP) in ontrol nd L-NAME - indued hypertensive rts. Systoli pressure (mm Hg) weeks Control Control+Borneol (50mg/kg) L-NAME (40mg/kg) L-NAME+Borneol (50mg/kg) L-NAME+Nifedipine (20mg/kg) d Vlues re given s men ± SD for six rts in eh group. Vlues not shring ommon supersript differ signifintly t p < 0.05 DMRT). d d 2010, IJPBA. All Rights Reserved. 274 Distoli pressure (mm Hg) weeks Control Control+Borneol (50 mg/kg) L-NAME(40mg/kg) L-NAME+Borneol (50 mg/kg) L-NAME+Nifedipine (20mg/kg) Vlues re given s men ± SD for six rts in eh group. Vlues not shring ommon supersript differ signifintly t p < 0.05 DMRT).

5 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts Tle 3.Effet of orneol on the tivity of SOD, tlse nd GPx in the erythroyte nd tissues of norml nd L- NAME indued hypertensive rts Groups Prmeter Smples Control Control + L-NAME L-NAME+ L-NAME + orneol (40mg/kg orneol (50 nifedepine (20 (50 mg/kg BW.) mg/kg BW.) mg/kg BW.) Erythroyte*/mg H) 6.88±0.92 d 7.29±0.76 BW ) 3.39± ±0.59 d 7.39±0.60 e SOD Liver (U*/mg 8.85± ± ± ± ±0.72 d Kidney(U*/mg 15.40± ± ± ± ±1.23 d Hert(U*/mg 5.99± ± ± ± ±0.58 d Erythroyte*/ mg ± ± ± ± ±9.11 d Ctlse H ) Liver(U*/mg 73.12± ± ± ± ±7.48 d Kidney(U*/mg 36.52± ± ± ± ±1.96 d Hert (U*/mg 50.96± ± ± ± ±5.04 d Erythroyte*/mg 14.31± ± ± ± ±0.22 H) GPx Liver(U*/mg 8.91± ± ± ± ±0.34 d Kidney(U*/mg 8.66± ± ± ±0.49 d 6.44±0.24 e Hert (U*/mg 8.41± ± ± ± ±0.32 d U* = enzyme onentrtion required to inhiit the hromogen produed y 50% in one minute under stndrd ondition. U # = µmole of H 2 O 2 onsumed/minute. U $ = µg of GSH utilized/minute.vlues re mens ± SD for six rts. Vlues not shring ommon supersript differ signifintly t p < 0.05 (DMRT) Tle 4. Effet of orneol on Vitmin-C, Vitmin-E nd GSH, in the plsm nd tissues of norml nd L-NAME indued hypertensive rts Groups Prmeter Vitmin C Vitmin E GSH Smples Control Control + orneol (50 mg/kg BW.) L-NAME (40mg/kg BW.) L-NAME+ orneol (50 mg/kg BW.) L-NAME+ nifedipine (20 mg/kg BW.) Plsm (mg/dl) 2.42± ± ± ± ±0.89 d Liver (µg/mg 0.83± ± ± ± ±0.05 d Kidney (µg/mg 0.84± ± ± ± ±0.10 d Hert (µg/mg 0.52± ± ± ± ±0.08 d Plsm (mg/dl) 2.62± ± ± ±0.11 d 1.72±0.16 Liver (mg/100 g wet 6.36± ± ± ± ±0.48 d Kidney(mg/100gwet 4.66± ± ± ± ±0.41 d Hert (mg/100 g wet 4.40± ± ± ± ±0.65 d Plsm (mg/dl) 34.84± ± ± ± ±1.74 Liver (mg/100 g wet 13.20± ± ± ±1.09 d 8.44 ±0.91 e Kidney(mg/100gwet 11.91± ± ± ± ± , IJPBA. All Rights Reserved. 275

6 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts Hert (mg/100 g wet 8.37± ± ± ± ±0.63 d Vlues re mens ± SD for six rts. Vlues not shring ommon supersript differ signifintly t p < 0.05 (DMRT) HISTOPATHALOGICAL EXAMINATION In ontrol, hert tissue smple showed onsistently norml histology (Fig. 3).Hert tissue smples were norml in ontrol, ontrol+orneol hypertensive rteriole hnges in L-NAME group (Fig.3C). Suh hnges were notly orneol treted nd nifedipine in L-NAME groups (Fig.3D groups (Fig.3A nd B).There were mild nd E). Fig.3.Representtive photomirogrph of histologil hnges in hert: (A) Control, myordil fires looked norml.(b) Control+orneol, similr to ontrol group.(c) L-NAME, there ws myordil fires destrution with mononuler ell infiltrtion, thikened rteriole medil hypertrophy.(d) L-NAME + orneol, vsulr hnges were milder thn L-NAME group minly in the form of mild mononuler inflmmtory ells seen in interstitum.(h&e prffin setion,5 µm thik, 40x ). A B C D E DISCUSSION Nitri oxide (NO) synthesis nd relese y endothelil ells ply n importnt vsulr relxtion effet ontriuting to the modultion of vsulr tone [23]. In ddition, NO hs een identified s importnt in other ellulr events, suh s vsulr smooth musle ell prolifertion nd neurl trnsmission. Chroni inhiition of NO produes volume-dependent elevtion of lood pressure nd its physiologil nd pthologil hrteristis resemle essentil hypertension. Besides, it is well estlished tht ute inhiition of nitri oxide iosynthesis y in vivo dministrtion of L-NAME, n L-rginine nlogue, leds to rteril hypertension nd renl vsoonstrition [24]. This effet is due to severl ftors like; inhiition of the NO prodution, inrese in symptheti tone, inrese in the rennin-ngiotensin system tivity [25] nd inrese in the vsulr resistne [26]. Also, the retivity to vsoonstritor gents is inresed nd the responsiveness to endothelil dependent vsodiltors is redued in this model of hypertension [27].Our results showed tht orl dministrtion of orneol signifintly ttenuted SBP nd DBP in L-NAME rts. Lipid peroxidtion is reported to e signifint ftor in pthologi onditions suh s hypertension, dietes mellitus nd myordil infrtion [28]. An inresed onentrtion of lipid peroxidtion end produts is notied in hypertension. To evlute the protetive role of orneol ginst hypertension-ssoited oxidtive stress, TBARS nd hydroperoxide levels in plsm nd tissues suh s liver, kidney nd hert were exmined in ll experimentl rts. In the present study, oth plsm nd tissue levels of TBARS nd hydroperoxides were enhned in L- NAME dministered rts s ompred to norml ontrol rts. L-NAME rts treted with orneol for 30 dys signifintly redued oth plsm nd tissue lipid peroxidtion levels. The ody hs evolved omplex defense strtegy to minimize the dmging effets of vrious oxidnts. Centrl to this defense, re the ntioxidnt enzymes of the lood (SOD, CAT nd GPx), whih t in onert to protet the orgnism from oxidtive dmge. SOD svenges the superoxide nion to form hydrogen peroxide, hene diminishing the toxi effet used y this rdil. The selenium-ontining enzyme GPx detoxifies H 2 O 2 y utilizing GSH nd H 2 O 2 s sustrtes to yield H 2 O nd oxidized glutthione [29]. Ctlse removes H 2 O 2 y reking it down diretly to O 2 [30]. In the present study, our results showed tht the tivities of SOD, CAT nd GPx were redued in erythroytes nd tissues of L-NAME dministered rts when ompred to ontrol rts. When orneol ws dministered, the redution in ntioxidnt enzyme tivities found in L - NAME rts ws reversed to vlues loser to those found in untreted, ontrol rts. The inresed tivities of SOD, CAT nd GPx ould e the result of deresed utiliztion 2010, IJPBA. All Rights Reserved. 276

7 Booln Rj et. l/ Antihypertensive nd Antioxidnt Potentil of Borneol-A Nturl Terpene in L-Nme Indued Hypertensive Rts eing tht the lipid peroxidtion ws low in the orneol-treted group. Severl studies hve suggested n inverse ssoition etween dietry intke nd plsm onentrtion of ntioxidnts nd vitmins nd rdiovsulr disese [31]. The derese in the levels of β-rotene, sori id nd α- toopherol my e the result of their inresed utiliztion to trp the ROS in L-NAME dministered rts ompred to the ontrol rts. Our results showed tht tretment with orneol inresed the onentrtion of β-rotene, sori id nd α-toopherol in L-NAME dministered rts. It hs een reported tht α- toopherol tretment lowers lood pressure in rts [32]. Similrly, severl ross-setionl studies reveled highly signifint inverse reltionships etween lood pressure nd sori id [33]. The onjugted system of et-rotene rets with peroxy rdils, thus reking hin retion of lipid peroxidtion [34]. The tripeptide GSH is one of the most importnt endogenous ntioxidnts. It plys the role of sulfhydryl group provider for diret svenging retion tlyzed y GPx nd s svenger of vitmin C nd vitmin E rdils. Tretment with orneol used n inresed genertion of glutthione in the liver of rts [35]. By inresing the levels of glutthione nd y deresing lipid peroxidtion, orneol my enhne resistne of ells memrne to freerdil medited injuries. Morphologil studies of the hert showed tht tretment with orneol minimized myordil fires destrution with mononuler ell infiltrtion, rteriolr thikening, glomeruloselrosis with mrked rteril hypotrophy hnges tht were hypertension, inresed tivity of the reninngiotension nd symptheti system nd signifint prodution of ROS nd inflmmtory meditors.. A vriety of terpenoids hve een shown to e effetive ginst oronry hert disese [36]. Reently, it hs een reported tht orneol hs n ntithromoti nd ntipltelet tivity in rts [13].This my e one of the reson for redution of lood pressure whih is notied in our study. Further, euse of its strong ntioxidnts properties, orneol svenging free rdils tht enhning levels of enzymi nd non-enzymi ntioxidnt nd exerts rdioprotetive properties. Therefore, we suggest tht dily onsumption of orneol might e effetive in lowering possile oxidtive dmge in hypertensive rts, inditing its therpeuti potentil s n ntihypertensive drug. In onlusion, NO prrine vsodiltor hs een implited in regulting vsulr tone nd myordil ontrtility nd n inhiitor of pltelet ggregtion ontriuting e ritilly in the development of hypertension. In our present study suggests tht orneol hve ntioxidnt nd ntihypertensive potentil effet in L-NAME indued hypertensive rts. REFERENCES 1. Yusuf S, Ounpuu S, Annd A. Glol urden of rdiovsulr diseses: Prt 1 Generl onsidertions, the epidemiologi trnsition, risk ftors nd impt of urniztion. Cirultion 2001;104: Knnel WB. Elevted systoli lood pressure s rdiovsulr risk ftor. Am J Crdiol 2000; 85: Mwmo ZH, Luyengi, Kinghorn AD. Phytohemils: glimpse into their struturl nd iologil vrition. Int J Phrm 1996;34: Mukherjee PK. Evlution of Indin trditionl mediine. Drug Informtion J 2001; 35: Komiy M, Tkeuhi T, Hrd E. Lemon oil vpor uses n nti-stress effet vi modulting the 5-HT nd DA tivities in mie. Behv Brin Res 2006;172: Grnger RE, Cmpell EL, Johnson GR. (+)- nd (-)-orneol: Effiious positive modultors of GABA tion t humn reominnt α1 β2 γ2l GABA A reeptors. Biohem Phrmol 2005; 69: Wng G, Wng L, Xiong ZY, Mo B, Li, TQ. Compound slvi pellet, trditionl Chinese mediine, for the tretment of hroni stle ngin petoris ompred with nitrtes: met-nlysis. Med Si Monit 2006; 12: Buhuer G, Jger W, Jirovetz L, Dietrih H. Effet of vlerin root oil, orneol, isoorneol,ornyl ette nd isoornyl ette on the motility of lortory nimls (mie) fter inhltion. Phrmol 1992;69: Wenqing G, Shufen L, Ruixing Y, Ynfeng H. Comprison of omposition nd ntifungl tivity of Artemisi rgyi Levl. et Vnt infloresene essentil oil extrted y hydrodistilltion nd superritil ron dioxide. Nt Prod Res 2006; 20: Cndn F, Unl M, Tepe B, Dferer D, Polissiou M, Smen A, Akpult HA. 2010, IJPBA. All Rights Reserved. 277

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