The Possible Protective Effect of Sage (Salvia Officinalis L.) Water Extract Against Testes and Heart Tissue Damages of Hypercholesterolemic Rats

Transcription

1 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts Fwzi A. Alshuily*, Etihj J. Jmi Biohemistry Deprtment, Fulty of Siene, King Adulziz University, Kingdom of Sudi Ari. ABSTRACT Sge (Slvi offiinlis) is one of the romtis nd mediinls hving iologil tivities nd phrmologil properties. The possile protetive role nd ntioxidnt tivity of sge wter extrt (SWE) ginst metoli disorders indued y hyperholesterolemi diet (HCD) in the hert nd testis tissues of rts were evluted in this study. The results of gs hromtogrphy/mss spetrometry (GC/MS) nlysis reveled tht Cmphor, 1,8-ineole nd α-thujone represent the min ompounds of voltile oil extrted from S. offiinlis. The findings of the iologil study showed tht the dietry intke of HCD signifintly inresed serum lipid ontents, tivities of rdi mrker enzymes nd the level of mlondildehyde (MDA) in testes nd hert tissues, nd signifintly redued the level of high-density lipoprotein-holesterol (HDL-C), testosterone (T), follile stimulting hormone (FSH) nd leutinizing hormone (LH) with suppression of ntioxidnt sttus ompred to the ontrolled rts. Wheres, supplementtion of rts with HCD long with SWE (1 ml/ Kg B. wt) redued the dmging effets indued y HCD with signifint derese in hyperholesterolemi stte, redution of lipid peroxidtion, indution of hert nd testes funtions nd elevtion of the tivity of ntioxidnt enzymes nd glutthione ontents. In onlusion, this study showed tht SWE ould e used s nturl nti-oxidnts supplements due to the presene of essentil oil, phenoli ontents nd other ntioxidnt omponents in Sge. Key Words: Slvi Offiinlis, Hyperholesterolemi Diet, Essentil Oil, Antioxidnt Ativity eijppr 2018; 8(1):62-68 HOW TO CITE THIS ARTICLE: Fwzi A. Alshuily, Etihj J. Jmi. (2018). The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts, Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh, 8(1), pp INTRODUCTION Hyperholesterolemi (HC) is one the mjor risky diseses tht hs een inresed in developing ountries in reent yers [1]. The ourrene of hyperholesterolemi indued over prodution of retive oxygen speies with elevtion of lipid peroxide ontent nd inhiition of ntioxidnt defense system [2]. Therefore, HC is losely ssoited with developing of mny hroni diseses inluding rdiovsulr diseses, progression of renl filure nd ertin metoli nd reprodutive disorders [3]. It hs een estimted y the World Helth Orgniztion (WHO) tht pproximtely 26 million deths our nnully euse of hyperholesterolemi-relted rdiovsulr diseses (CVDs) [4]. Severl lipids lowering drugs suh s firtes, sttins, nd ile id sequestrnts n e used to regulte the lipid metolism y different mehnisms ut these drugs hve mny side effets [5]. There hs een n interest in n lterntive mediine in reent dedes, euse there re thousnds of potentil mediinl plnts tht ontin mny iotive sustnes nd ntioxidnts [6]. Sge (Slvi offiinlis L., Lmiee) is ommon romti plnt tht hs een used s mediinl plnt in treting different diseses suh s ner, dietes, hert diseses, reprodutive disorders nd Corresponding uthor: Fwzi A. Alshuily Address: Biohemistry Deprtment, Fulty of Siene, King Adulziz University, Kingdom of Sudi Ari E-mil: flshili@ku.edu.s Relevnt onflits of interest/finnil dislosures: The uthors delre tht the reserh ws onduted in the sene of ny ommeril or finnil reltionships tht ould e onstrued s potentil onflit of interest. Reeived: 01 Novemer 2017; Revised: 25 Jnury 2018; Aepted:12 Ferury2018

2 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts hyperholesterolemi [6]. The enefiil effets of Sge hve een relted to its iotive omponents inluding voltile ftty ids, diterpens, sponins, flvonoids, phenoli ids, resin nd oestrogeni sustnes. With regrd to the possile ntioxidnt tivity of sge, the im of this study ws to evlute the protetive effets of Slvi offiinlis ginst metoli disorders indued y hyperholesterolemi diet in the hert nd testes tissues of rts. MATERIALS AND METHODS Dried leves of Sge were purhsed from lol her mrket. Chemils nd regents were purhsed from Sigm Chemil Co. (St. Louis, MO, USA). Anlysis of sge essentil oil y GC/MS: The essentil oil ws isolted from sge (100g) ording to the method of [7]. The voltile oil nlysis ws rried out using gs hromtogrphy-mss spetrometry instrument GC/MS. Preprtion of sge wter extrt (SWE): Sge wter extrt (sge te): It ws prepred y pouring 150 ml oiling wter onto 2 g of dried grounded leves nd llowing it to steep for 5 min [8]. Experimentl Protool: Animls nd iohemil Assy Mle lino rts (150 ± 20 g) were limted to ontrolled lortory onditions for two weeks. Rts were mintined on stok rodent diet nd tp wter tht were llowed d liitum. The rodent ontrolled diet ws omposed of 15% sein, 10% orn oil, 5% ellulose, 4% slt mixture, 1% vitmins mixture nd strh 65% [9]. All the niml proedures were omplished sed on the Ethis Committee of the Ntionl Reserh Centre onformed to the Guide for the Cre nd Use of Lortory Animls, pulished y the US Ntionl Institutes of Helth (NIH pulition No , 1996). Experimentl design The nimls were rndomly divided into four groups, eh omprised of 7 rts. Group C: rts were fed on norml diet for 10 weeks, served s ontrol, Group HCD: rts were fed on hyperholesterolemi diet (sl diet + 1% holesterol+ 16% ft nd 0.2% Choli id) [10] for 10 weeks. Group SWE: rts were fed on norml diet nd supplemented orlly with sge wter extrt (1 ml/ Kg ody wt) [11] for 10 weeks. Group HCD + SWE: rts were fed on HCD supplemented orlly with sge wter extrt (1 ml/ Kg B. wt) [11] for 10 weeks. Eh rt ws weighed t the initil nd end of the experiment. At the end of the experiment (10 weeks), rts were fsted for 24 hours nd nesthetized with diethyl ether. For otining serum for iohemil nlysis, lood smples were olleted through hert punture nd llowed to ogulte, nd then entrifuged. Also, for iohemil investigtion, rdi nd testes tissues were disrded. Biohemil Anlysis Totl holesterol (TC), triglyerides (TG) nd high-density lipoprotein-holesterol (HDL-C) were inditd sed on the proedure desried y [12], [13], nd [14], respetively. Low-density lipoprotein-holesterol, very Low-density lipoprotein-holesterol nd risk rtio were evluted ording to [15] y the following equtions: LDL-C (mg/dl) = TC-(TG/5+HDL-C), vldl (mg/dl) = TG/5. The levels of ltte dehydrogense (LDH) nd retine phosphokinse (CPK) were determined y the method of [16], sprtte trnsminse (AST) nd lnine trnsminse (ALT) were determined ording to [17]. Estimtion of testosterone hormone ws performed sed on the method of [18], Follile stimulting hormone (FSH) nd leutinizing hormone (LH) ording to [19]. Crdi nd testes tissues were disseted, thoroughly wshed with ie-old 0.9% NCl, weighed, mined nd homogenized (10% w/v) using 66 mmol/l hilled phosphte uffer (ph 7.0). The tissue homogentes were entrifuged t 6000 rpm for 15 min nd the superntnts were used to estimte mlondildehyde (MDA) [20], GSH [21], superoxide dismutse tivity (SOD) [22] nd Ctlse tivity (CAT) [23]. Sttistil nlysis Results were presented s men ± SE (n = 7). Experimentl dt were nlyzed using one-wy nlysis of vrine (ANOVA). Dunn s multiple rnge test ws used to determine signifint differenes etween mens. The sttistil nlyses were performed using omputer progrm Sttistil Pkges for Soil Siene (SPSS) [24]. Differenes etween mens were onsidered signifint t P < RESULTS The results of GC/MS nlysis showed tht S. offiinlis is omposed of high mount of essentil oils, most of them re romti. Cmphor (24.55%), 1,8-ineole (17.86%) nd α-thujone (14.07%) re represented s the mjor essentil oils of sge (Tle 1). 63

3 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts Tle 1. Perentge of essentil oil onstituents of Slvi offiinlis (%) Compound Perentge (%) Compound Perentge (%) limonene 3.61 α-terpinyl ette ,8-ineole γ-selinene 3.54 linlool 0.19 ryophyllene oxide 1.12 α-thujone humulene epoxide 1.09 β- thujone 6.03 isoromdendrene epoxide 0.35 β-pinene 0.23 trns-z- α- isolene epoxide 0.21 ryophyllene 3.92 Cryophyleine (I3) 1.16 mphor mnoyl oxide 5.57 humulene 4.73 β-selinene 0.39 α-terpineol 1.37 orneol 3.13 Totl identified ompounds Dt showed tht HCD-group hd signifint inrese in the serum level of lipid ontents (TC, TG, LDL-C nd vldl-c) nd remrkle derese in HDL-C reltive to the ontrol group. On ontrst, supplementtion of rts with HCD long with SWE indued signifint inrese in HDL-C nd redution in the serum level of TC, TG, LDL-C nd vldl-c when ompred to the rts fed on HCD only (Tle 2). Tle 2. Effet of dministrtion of sge wter extrt on serum lipid profile in diet indued hyperholesterolemi rts TC TG HDL-C LDL-C v-ldl Animl groups (mg/dl) (mg/dl) (mg/dl) (mg/dl) (mg/dl) Control ± ± ± ± ±2.64 HCD ± ± ± ± ±7.82 SWE ± ± ± ±2.11 d 30.74±2.33 HCD+ SWE ± ± ± ± ±3.64 Mens in the sme olumn with different supersripts re signifintly different t (P<0.05), Vlues re expressed s men ± S.E. (n=7) HCD: Hyperholesterolemi diet, SWE: Sge wter extrt. 64 The results indited tht HCD indued signifint inrese in the tivity of serum LDH, CPK, AST nd ALT ompred with the ontrol group. The tivity of these enzymes signifintly redued in the group of rts whih reeived HCD nd SWE when ompred to the rts reeived HCD (Tle 3). Tle 3. The Effet of dministrtion of sge wter extrt on serum LDH, CPK, ALT nd AST tivities in diet indued hyperholesterolemi rts Animl groups LDH (nmol/ml) CPK (nmol/ml) ALT (U/L) AST (U/L) Control ± ± ± ±1.57 HCD ± ± ± ±2.92 SWE ± ± ± ±1.66 HCD+ SWE ± ± ± ±2.15 Mens in the sme olumn with different supersripts re signifintly different t (P<0.05), Vlues re expressed s men ± S.E. (n=7) HCD: Hyperholesterolemi diet SWE: Sge wter extrt

4 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts In this work, the indution of hyperholesterolemi in rts resulted in remrkle redution in the level of testosterone, LH nd FSH ompred to the ontrolled rts. On the other hnd, feeding rts HCD with SWE indued signifint inrese in the level of these hormones ompred to HCD group (Tle 4). Tle 4. The effet of dministrtion sge wter extrt on the level of T, LH nd FSH in diet indued hyperholesterolemi rts Animl groups T (n mol/l) LH (IU/L) FSH (IU/L) Control 4.80± ± ± 0.05 HCD 2.42± ± ± 0.04 SWE 4.91± ± ± 0.06 HCD+ SWE 3.96± ± ± 0.05 Mens in the sme olumn with different supersripts re signifintly different t (P<0.05), Vlues re expressed s men ± S.E. (n=7) HCD: Hyperholesterolemi diet SWE: Sge wter extrt The rdi nd testiulr tissues of HC-rts exhiited signifint inrese in the level of MDA nd signifint derese in the level of GSH nd tivity of the enzymti ntioxidnts (CAT nd SOD) ompred to those from the ontrol group. By ontrst, in the HCD nd SWE group, MDA signifintly redued nd GSH nd the ntioxidnt enzymes signifintly improved (Tle 5). Tle 5. The Effet of dministrtion of sge wter extrt on the rdi nd testiulr ntioxidnt sttus in diet indued hyperholesterolemi rts Prmeters Control HCD SWE HCD+SWE MDA (n mol/g tissue) Hert ± ± ± ± ± ± ± ±4.73 GSH (mg/g tissue) Hert ± ± ± ± ± ± ± ±1.62 SOD (U/mg protein) Hert ± ± ± ± ± ± ± ±1.12 CAT (U/g protein) Hert 3.72± ± ± ± ± ± ± ±0.12 Mens in the sme row with different supersripts re signifintly different t (P<0.05), Vlues re expressed s men ± S.E. (n=7). HCD: Hyperholesterolemi diet. SWE: Sge wter extrt DISCUSSION Consumption of HCD ould e linked to the metoli disorders nd hroni diseses nd n led to weight gin nd elevted lood lipid levels [25]. In this reserh, the protetive role of the SWE on the hert nd testes tissues of hyperholesterolemi rts hs een emphsized. The results of GC/MS nlysis of the essentil oil of sge in this study were in greement with the results presented in the study of [26] nd [27] ut with vrile onentrtions depending on lotion nd stges of development, environmentl, physiologil nd morphologil ftors [28]. The oil omposition ws found to meet the stndrds given y ISO 9909 for use of S. offiinlis for mediinl purposes [29]. [27] otined tht the oil omposition of S. offiinlis ws found to e rih in oxygented monoterpenes inluding α-thujone, β-thujone, 1,8-ineole, mphor nd orneol. In this work, hyperholesterolemi in rts ws indued y feeding rts HCD for 10 weeks. The levels of serum TC, TG, LDL-C nd vldl-c signifintly inresed ove the norml upper limits nd the level of HDL-C redued signifintly under the effet of HCD reltive to the norml group. HCD ws onerned with the inresing onentrtions of serum nd hepti totl TC, espeilly the level of vldl-c nd LDL-C in serum, whih is onsidered to e primry risk ftor of rdiovsulr disese [30]. Also, [30] demonstrted tht the holesterol dministrtion redued the ftty id tolism, whih my ount for the enhnement of hepti TG level. A signifint rise in the tivity of rdi enzymes LDH, 65

5 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts CPK, AST nd ALT ws oserved in the group HCD when ompred to the ontrolled rts. The elevtion in the tivity of these enzymes ould e ttriuted to the tissue dmge indued y HC nd the lekge of these mrkers in the plsm [31]. Furthermore, HCD ould indue suppression in hepti nd rdi funtions evidened y ugmenttion of serum levels of AST, ALT, LDH nd CK- MB tivities [32]. The dt in this work reveled tht HCD uses reprodution prolems y deresing the level of testosterone, LH nd FSH ompred to the ontrolled rts. [33] indited tht the high ft diet indued n inrese in totl holesterol nd triglyerides levels nd the poor development of testiles in puertl rts. Hyperholesterolemi uses disturne in hypothlmus pituitry xis tivity. [34] reported tht the inrese of oxidtive stress nd derese of endogenous ntioxidnt tivity led to the spermtogenesis disturne. The indution of HC in this study resulted in signifint inrese in oxidtive stress in hert nd testes tissues, evidened y high level of MDA nd signifint derese in the level of GSH nd tivity of the enzymti ntioxidnts (CAT nd SOD) ompred to those from the ontrol group. [35] suggested tht hyperholesterolemi leds to the inresed lipid peroxidtion whih is in onsistent with severl linil nd experimentl studies. Oxidtive stress hs emerged s n importnt pthogeni ftor in the development of hypertension nd lso most of the omplitions relted to the hypertension re ssoited with oxidtive stress, indued y the genertion of free rdils [36]. [37] indited tht free rdil nd suppression of ntioxidnt enzymes re the prinipl ftors involved in the pthogenesis of hyperholesterolemi. [38] onluded tht hyperlipidemi indued y diet redues the SOD, CAT, GPx nd GST tivities, nd prllely elevte the lipid peroxide levels, therey leding to inhiition of ntioxidnt defense system. On the other side, feeding rts HCD with SWE resulted in n ovious protetion of SWE ginst the disturne indued y HCD evidened y remrkle redution in the lipid profile ontents, tivity of serum LDH, CPK, AST nd ALT nd lipid peroxidtion ompnied y signifint elevtion in the HDL-C, testosterone, LH nd FSH nd tivtion of ntioxidnt defense system. The study of [39] reveled tht Slvi offiinlis te onsumption is ountle for the improvement of the lipid profile induing derese on the highly therogeni LDL-C prtiles nd n inrese in the HDL-C whih my e due to the ility of SWE to redue holesterol iosynthesis. The hypoholesterolemi effet of SWE my e ttriuted to severl sge nturl omponents tht hve een shown to t on holesterol metolism y reduing its sorption or its synthesis, suh s tehins [40]. Also, thujone, one of monoterpenes tht ws found s mixture of lph nd β distereoisomers in Slvi offiinlis L., lowers holesterol nd triglyeride levels [41]. The effet of SWE on rdi enzymes in this study is in greement with the results of [42] who indited tht the tretment of methomyl-dministered rts with Slvi offiinlis nd Rut grveolens ethnoli extrts produed potentil meliortion of the impired CK-MB, AST nd LDH tivities. The uthors reveled tht the ntioxidnt tivity of S. offiinlis n protet ginst ellulr dmge indued y degenertive diseses nd therey prevent lekge of rdi enzymes in the plsm [42]. [43] onluded tht the queous extrt of Slvi offiinlis used positive effet on some fertility prmeters nd pituitry-testiulr hormone xis y inresing the level of Testosterone, LH nd FSH. The improving effets of SWE on mle reprodutive system my e relted to the effets of Sge omponents inluding vitmins C nd E, flvonoids nd ntioxidnt [43]. [44] reported tht Slvi offiinlis extrt hs the ility to stimulte the growth of testes nd enhne the prolifertion, mturtion nd differentition of spermtozo due to the presene of sponine, lkloids in the slvi extrt. It ws found tht the ntioxidnt properties of SWE ould e ttriuted to the presene of lot of tive omponents tht hve high ntioxidnt tivity suh s rnosol, rnosi nd rosmrini ids, rosmdil, rosmnol, epirosmnol, methyl rnoste, nd luteolin-7-o-β-gluopyrnoside. These ntioxidnt ompounds hve the ility to stimulte endogenous ntioxidnt defense systems nd svenge retive speies [45]. [46] suggested tht the presene of vitmins C nd E, flvonoids nd phenoli ompounds in SWE might led to the regultion of signl trnsdution pthwy of ell growth nd prolifertion, modultion of enzyme tivity relted to the detoxifition, stimultion of the immune system nd DNA repir nd regultion of hormone metolism. CONCLUSIONS From the results of this study, it ould e postulted tht SWE plys vitl role ginst HC nd indues the repir of the dmged hert nd testes. SWE redues the level of lipid ontents, inreses ellulr nti-oxidnt tivities nd protets rdi nd testiulr tissues y svenging free rdils. This reserh onluded tht the iologil tivity of SWE ould e ttriuted to the presene of the essentil oil, phenoli ontents nd other ntioxidnt omponents. Therefore, SWE ould form promising resoures to the supplement for nturl nti-oxidnts. REFERENCES [1] Chng C-J, Lu C-C, Lin C-S, Mrtel J, Ko Y-F, Ojius DM, Wu T-R, Tsi Y-H, Yeh T-S, Lu J-J, Li H-C nd Young JD (2018): Antrodi innmome redues oesity nd modultes the gut miroiot in high-ft diet-fed mie. Interntionl Journl of Oesity 42: [2] Akinyemi, A. J., Ooh, G., Ademiluyi, A.O., Boligon, A. A. nd Athyde M.L. (2016). Effet of two ginger vrieties on rginse tivity in hyperholesterolemi rts. J Aupunt Meridin Stud, 9(2): [3] Cmpos-Silv P., Furriel A., Cost W. S., Smpio F.J. B., Gregório B. M. (2015). Metoli nd testiulr effets of the long-term dministrtion of 66

6 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts different high-ft diets in dult rts. iju 41 (3): [4] Khn T. J., Ahmed Y. M., Zmzmi M. A., Mohmed S. A., Khn I., Bothmn O. A. S., Mehnn M. G. nd Ysir M. (2018). Effet of torvsttin on the gut miroiot of high ft diet-indued hyperholesterolemi rts SCIentIFIC Reports. 8:662. [5] Golkrishnn S., Asirvthm S. S. nd Jnrthnm V. (2016). Effet of Silyin on Lipid Profile in Hyperholesterolemi Rts. Journl of Clinil nd Dignosti Reserh. 10(4): FF01-FF05 [6] Ben Khedher M.R., Hmmmi M., Arh J. R.S., Hislop D. C., Eze D., Wrgent E.T., Kępzyńsk M. A. nd Zi M. S. (2018). Preventive effets of Slvi offiinlis lef extrt on insulin resistne nd inflmmtion in model of high ft diet-indued oesity in mie tht responds to rosiglitzone. PeerJ 6: e4166; DOI /peerj.4166 [7] Gunther G. (1961). The Essentil Oils. Nstrnd Press, New York, USA. [8] Osmn N. N. nd Ad El Azime A. Sh (2013). Slvi offiinlis l. (sge) Ameliortes Rdition-Indued Oxidtive Brin Dmge in Rts Ar Journl of Nuler Siene nd Applitions. 46(1): [9] Philip, G.R., Forrest, H.N. nd George, C.F. (1993). AIN 76A rodent diet. J of Nut. 123: [10] Hrnfi H., Aziz M. nd Amrni S. (2009). Sweet sil (Oimum silium L.) improves lipid metolism in hyperholesterolemi rts. Spn the Europen J. of Clin. Nut. nd Metolism. 4: e181-e186. [11] Ad El Ftth S. M., El-Ftih N. M. nd Fhim Th. M. (2013). Effet of Using Aqueous Extrt of Slvi offiinlis L. Leves on Some Antioxidnts Sttus in Irrdited Rts Egypt. J. Rd. Si. Appli. 26: [12] Allin C. C., Poon L. S., Chn C. S., Rihmond W. nd Fu, P. C. (1974). Enzymti determintion of totl serum holesterol. Clin. Chem. 20: 470. [13] Fossti, P. nd Prenipe, L. (1982). Serum triglyerides determined lorimetrilly with n enzyme tht produe hydrogen peroxide. Clin. Chem., 28:2077. [14] Demker, P. N., Vos-Jnssen, H. E., Hifmns, A. G. M., Vn't Lr, A. nd Jnsen, A. P. (1980). Mesurement of high- density lipoprotein holesterol in serum: Comprison of six isoltion methods omined with enzymti holesterol nlysis. Clin. Chem., 26:1780. [15] Friedwld, W.T., Levy, R. I. nd Fredrikson, D. (1972). Estimtion of onentrtion of low-density lipoprotein holesterol in plsm without use of the preprtive ultrentrifuge. Clin Chem. 18(6):499. [16] King J (1965) (ed). The dehydrogenses or oxidoredutses - ltte dehydrogense. In: Prtil Clinil Enzymology. Vn Nostrnd Compny Ltd, London, [17] Reitmn S. nd Frnkel, S. (1957). A lorimetri method for the determintion of serum glutmi oxleti nd glutmi pyruvi trnsminses. Am. J. Clin. Pthol. 28: 56. [18] Wilson J.D. nd Foster D.W. (1992). Willims Textook of Endorinology. Phildelphi: Sunders, [19] Grrett, P.E. (1989). The enigm of stndrdiztion for LH nd FSH ssys. J Clin Immunossy 12: [20] Yoshiok, T., Kwd, K., Shimd, T. nd Mori, M. (1979). Lipid peroxidtion in mternl nd ord lood nd protetive mehnism ginst tivted-oxygen toxiity in the lood. Am J Ostet Gyneol. 135: [21] Beutler, E., Duron, O. nd Kelly, B.M. (1963). Improved method for the determintion of lood glutthione. J L Clin Med. 61: [22] Minmi, M. nd Yoshikw, H. (1979). A simplified ssy method of superoxide dismutse tivity for linil use. Clin. Chim. At., 92: [23] Johnsson L. H. nd Borg L.A.H. (1988). A spetrophotometri method for determintion of tlse tivity in smll tissue smples. Anl. Biohem. 74: 331. [24] SPSS. (1998). Sttistil Pkge for Soil Siene. Computer Softwre, Ver. 10. SPSS Compny, London, UK. [25] Singh A., Gowthm S., Chkrpni L.N., Ashokkumr S., Kishore Kumr S.N., Prem V., Divy Bhvni R., Mohn T. nd Sthymoorthy Y. K. (2018). Aegeline vs Sttin in the tretment of Hyperholesterolemi: A omprehensive study in rt model of liver stetosis. Funtionl Foods in Helth nd Disese 8(1): [26] Mokute, D., Nivinskiene, O., Bernotiene, G. nd R. Butkiene (2003). The is-thujone hemotype of Slvi offiinlis L. essentil oils. Chemij (Vilnius). 14 (4): [27] Sid-Al Ahl H., Hussein M. S., Gendy A.S. H. nd Tkhenko K. G. (2015): Qulity of Sge (Slvi offiinlis L.) Essentil Oil Grown in Egypt. Interntionl Journl of Plnt Siene nd Eology. 1(4): [28] Couldis M., Tzkou O., Mimi-Dukić N., Jnčić R. nd Stojnović D. (2002). Essentil oil of Slvi offiinlis L. from Seri nd Montenegro. Flvour Frgr. J. 17: [29] Berotiene, G., Nivinskiene, O., Butkiene, R. nd D. Mokute (2007). Essentil oilomposition vriility in sge (Slvi offiinlis L.). Chemij, 18: [30] Wng Y-M, Zhng B., Xue Y., Li Z-J, J-F Wng, Xue C-H nd Yngit T. (2010). The mehnism of dietry holesterol effets on lipids metolism in rts. Lipids in Helth nd Disese 9:4. [31] Mitni, H., Egshir, K. nd Kimur, M. (2003). HMG-CoA redutse inhiitor, fluvsttin, hs holesterol lowering independent diret effets on therosleroti vessels in high holesterol diet-fed rits. Phrmol Res, 48: [32] Sunrunswt, T., Ayutthy, W. D. N., Songsk, T., Thirwrpn S., nd Poungshompoo S. (2011). Lipid-Lowering nd Antioxidtive Ativities of Aqueous Extrts of Oimum sntum L. Leves in Rts Fed with High-Cholesterol Diet. Oxidtive 67

7 Interntionl Journl of Phrmeutil nd Phytophrmologil Reserh (eijppr) Ferury 2018 Volume 8 Issue 1 Pge Fwzi A. Alshuily, The Possile Protetive Effet of Sge (Slvi Offiinlis L.) Wter Extrt Aginst nd Hert Tissue Dmges of Hyperholesterolemi Rts Mediine nd Cellulr Longevity Volume 2011, Artile ID , 9 pges. [33] Wng Y, Liu XP, Qin DN, Chen S nd Li YS (2007). Diet-indued oesity ffets testis development in puertl rts. Zhonghu Nn Ke Xue 13: [34] Bshndy A.E.S. (2007). Effet of fixed oil of Nigell stiv on mle fertility in norml nd hyperipidemi rts. Int J Phrm, 3 (1): [35] Dutt, K. nd Bishyi, B. (2009). Esherihi oli lipopolyshride dministrtion lters ntioxidnt profile during hyperholesterolemi. Indin J. Clin. Biohem. 24: [36] Sonker, R., Nidu, M., Rju, S.B., Prsd, A.K. nd Ro, T.R.K. (2012). Effet of et-1-loker, neivolol, on entrl orti pressure nd rteril stiffness in ptients with essentil hypertension. Indin J. of Phrmo., 44: [37] Yilmz MI, Bykl Y, Kili M, Sonmez A, Buluu F, Aydin A, et l (2004). Effets of sttins on oxidtive stress. Biol Tre Elem Res. 98: [38] Fki I., Bouziz M., Shnoun Z. nd Sydi S. (1997). Hyperholesterolemi effets of phenoli rih extrts of Chemlli olive ultivr in rts fed holesterol-rih diet. Bioorg Med Chem. 378: [39] Elid, B., Dniel, Z., Pyl, P., Vishl, J., Tejs, L., Inn, K., Sidney, J. nd Sidhrth, D. (2010). A novel dietry supplement ontining multiple phytohemils nd vitmins elevtes heptorenl nd rdi ntioxidnt enzymes in the sene of signifint serum hemistry nd genomi hnges. Oxid. Med. Cell Longev. 3 (2): [40] Pln, N., Niolle, C., Ferre, R., Cmps, J., Cós, R., Villori, J. nd Msn, L. (2008). DANACOL group. Plnt sterol-enrihed fermented milk enhnes the ttinment of LDLholesterol gol in hyperholesterolemi sujets. Eur. J. Nutr., 47: [41] Kee, E., Livengood, M., Crter, E., Mkenn, M. nd Cfiero, M. (2009). Aromti intertions in the inding of lignds to HMGCoA redutse. J. of Physis. nd Chem.,113 (44): [42] Ahmed O. M., Ashour M. B., Asrn A. E-M. A. nd Ali M. A. (2017). Assessment of the Preventive Effets of Slvi offiinlis nd Rut grveolens Ethnoli Extrts on Chlorpyrifosnd Methomylindued Testiulr nd Crdi Toxiities in Alino Rts Amerin Journl of Mediine nd Medil Sienes 7(7): [43] AL-Chli S., M.M, Shukri H. M., Mhmood R.I.nd Khlid L. B. (2016). Effet of Slvi offiinlis L. (Sge) Aqueous extrt on Liver nd Testiulr Funtion of Dieti Alino Mle Rts. Journl of Bylon University/Pure nd Applied Sienes. 24 (2): [44] Lim, C. F nd Femndes-Ferreir M. (2007). Drinking of Slvi offiinlis te inreses CCl4 idued heptotoxiity in mie. food nd hemil Toxiology., 45(3): [45] S C, Rmos A, Azevedo M, Lim C, Fernndes- Ferreir M nd Pereir-Wilson C. (2009). Sge te drinking improves lipid profile nd ntioxidnt defenes in humns. Int J Mol Si. 10: [46] Okenfull, D. (1996). Sponins in the tretment of hyperholesterolemi. In: Spiller GA. (ed.) Hndook of Lipids in Humn Nutrition. CRC Press, Bo Rton