Hypolipideimic and Hypocholestermic Effect of Pine Nuts in Rats Fed High Fat, Cholesterol-Diet

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1 Worl Applie Sienes Journl 15 (12): , 2011 ISSN IDOSI Pulitions, 2011 Hypolipieimi n Hypoholestermi Effet of Pine Nuts in Rts Fe High Ft, Cholesterol-Diet 1 2 Amr, A. Rezq n Aeer, E. El-Khmisy 1 Deprtment of Nutrition n Foo Siene, Fulty of Home Eonomis, Helwn University, Ciro, Egypt 2 Deprtment of Nutrition n Foo Siene, Fulty of Speifi Eution, Port-Si University, Port-Si, Egypt Astrt: Pine nuts re holesterol-free n goo soure of nutrients. It is rih in energy n onsists of protein, rohyrtes, ftty is, vitmins n minerls. The present stuy ws rrie out to investigte the hypolipieimi n hypoholestermi effet of pine nuts on rts fe high ft, holesterol-iet. Rts were ivie into five groups of seven rts eh. Group (1) kept s negtive ontrol group; the remining four groups fe high ft, holesterol iet. Group (2) kept s positive ontrol group; groups (3), (4) n (5) fe iets supplemente with 5, 10 n 15% pine nuts, respetively. Dt illustrte tht positive ontrol group h signifint reution in foo intke n no signifint hnge in oy weight gin. It lso h signifint inrese in serum levels of totl lipi (TL), triglyeries (TG), totl holesterol (TC), LDL-C, VLDL-C, AST, ALT n ALP n h signifint erese in serum level of HDL-C n vlue of HDL-C/TC rtio, ompre to the negtive ontrol group. In ontrst, supplemente iet with pine nuts use signifint reution in foo intke n non signifint hnge in oy weight gin. In ition to, signifint erese in serum levels of the ove mentione prmeters s well s signifint inrese in serum level of HDL-C n vlues of HDL- C/TC rtio, exept low level of pine nuts (5%) inue no signifint hnge in TG n HDL-C, s ompre the positive ontrol group. Norml histologil struture ws oserve in hert of trete rts with 10 n 15% pine nuts n in ort of trete rts with 15% pine nuts. It is onlue tht pine nuts inrese the reution in lipi profile, lipoprotein holesterol n liver enzymes. These ereses were inrese with inrese pine nut level. Key wors: Rts Hyperholesterolemi Hypertriglyeriemi Serum lipoproteins Pine nuts INTRODUCTION lumpe together in the ommere n referre to s pine nuts [3]. Pinus pine trees (Pinee fmily) re grown Pine nuts re holesterol-free [3] n re goo wiely in Europe n North Ameri. Pine nuts re the soure of nutrients. It is rih in energy (628kl) n eile sees of pines. Aout 20 speies of pine proue onsists of protein (11.6 g/100g), rohyrtes (19.3 sees lrge enough to e worth hrvesting; in other g/100g) n ftty is (61 g/100g) [4]. The most pines the sees re lso eile, ut re too smll to e of unnt mino is in pine nuts re tryptophn n gret vlue s humn foo [1]. Pine nuts re eten rw histiine s ompre to eggs [5]. Moreover, ftty is or roste; they re inlue s ingreients in vriety of omposition of pine nuts re 85% unsturte ftty is tritionl ishes suh s met, fish n vegetle ish n 15% sturte ftty is. The most ommon s well s hooltes n esserts [2]. Different pine nuts unsturte ftty is re linoli (48.4%), olei (25.50%) otine from vrious speies iffer in size, nutritionl / n pineloi (14.90%) is [6]. Pine nuts oil possess meiinl vlue n tste. However, onsumers re wie therpeuti effet, suh s ntiteril, ntifungl, usully not sophistite enough to istinguish etween ntivirl, ntisepti, ntineurlgi, holgogue, holereti, nuts of ifferent speies, therefore the nuts re usully iureti, expetornt, hypertensive [7, 8]. Pine nuts lso, Corresponing Author: Amr A-Elmory Rezq, 65 St. Elmt Elhli, Bolq Au Ell, Fulty of Home Eonomis, Helwn University, Ciro, Egypt. 1667

2 Worl Appl. Si. J., 15 (12): , 2011 ontin vitmins, prtiulrly B1 n B2 n minerls, espeilly potssium n phosphorus. Aprt from nutritionl vlue, onsumption of nuts is helth. The omposition of pine nuts shows vrition mong speies n even some suspeies epening on geogrphil n limti onitions [9, 10]. Criovsulr isese is the most ommon use of eth in inustrilize ountries, with high plsm triglyerie (TG) levels eing suggeste to e inepenent risk ftor [11]. It is known, tht plsm levels of TG re influene y ietry omposition, smoking, oy weight n geneti ftors. Similrly to the other risk ftors, it is estimte tht the ontriution of geneti n environmentl ftors on plsm levels of TG is roughly the sme [12]. Hyperholesterolemi is prolem isese for mny soieties n is one of the mjor risk ftors for the evelopment of riovsulr iseses, suh s theroslerosis n its omplitions, ute infrttion of the myorium or hypertension [13-15]. In ition, there is orreltion etween these iseses n lipi normlities, espeilly high level of plsm holesterol n loo pressure [16]. Although the fous of reserh so fr hs een minly on the vsulr effets of hyperlipiemi, i.e. rterioslerosis, it is now quite evient tht hyperlipiemi exerts iret effets on the myorium in ition to the evelopment of theroslerosis [17, 18]. The known lipi lowering rugs (firtes, sttins, ile i sequestrnts, et.) regulte the lipi metolism y ifferent mehnisms, ut lso hve mny sie effets in ptients [19]. Therefore, the present stuy ime to investigte the hypolipieimi n hypoholestermi effet of pine nuts on rts fe high ft, holesterol-iet. This ws hieve y mesuring the serum onentrtion of totl lipi, triglyeries, totl holesterol n its lipoprotein frtions, AST, ALT n ALP, s well s histopthologil exmintion of hert, ort n liver. MATERIALS AND METHODS Rts n Diet: Thirty five mle Sprgue-Dwley rts weighing 200 ± 5 g were purhse from the Lortory Animl Colony, Ministry of Helth n Popultion, Helwn, Egypt. Cholesterol n sl iet onstituents were otine from El- Gomhory Compny for Chemil n Phrmeutil, Ciro, Egypt. Pine Nuts: Pine nuts were otine from lol mrket, Ciro Egypt. Chemils: Kits for iohemil nlysis of serum totl lipis, triglyeries, totl holesterol, HDL-C, AST, ALT n ALP were purhse from the Gmm Tre Compny for Phrmeutil n Chemils, Dokki, Egypt. Preprtion of Bsl Diet: The sl iet (AIN-93M) ws prepre oring to Reeves et l. [20]. Diet ws formulte to meet the reommene nutrients levels for rts. Experimentl Design: The experiment ws onute on thirty five mle rts weighing pproximtely 200 ± 5 g for 6 weeks. The nimls were house in helthy onition t room temperture (21-23 C), with 40-60% humiity, expose to 12:12-h light-rk yle n fe on the sl iet n wter ws provie liitum for one week efore strting the experimentl for limtiztion. After limtiztion perio rts were ivie into five groups of seven rts eh: group (1) fe the sl iet n kept s negtive ontrol group (norml rts); the remining four groups fe high ft, holesterol ontining - iets. Group (2) kept s positive ontrol group; groups (3), (4) n (5) fe supplemente iets with 5, 10 n 15% pine nuts, respetively. Hyperlipiemi n hyperholestermi in rts ws one oring to the metho of Blkn et l. [21]. In riefly, sl iet ws formulte with 1% holesterol, 2% sheep ft n 0.5% holi i to enhne the enterl sorption of lipis. At the en of the experimentl perio (6 weeks), iets were withhel from experimentl rts for 12-h n then rts were srifie. Bloo smples were ollete from the portl vein into ry len entrifuge tues. For serum seprtion, loo smples were left t room temperture to get lot n then entrifuge for 15 minutes t 3000 rpm. Serum ws refully spire using neele n trnsfers into ry len test tues n kept frozen t -10ºC until hemil nlysis. Orgns suh s liver, kiney n hert were remove n wshe with sline solution, rie n then weighte to lulte reltive orgns weight to oy weight. Hert, ort n liver of ll rts were kept in formlin solution (10%) for histopthologil exmintion. Foo Intke n Boy Weight Gin Assy: Foo intke (FI) ws lulte every other y. The iologil vlue of the ifferent iets ws ssesse y the etermintion of its effet on oy weight gin (BWG) t the en of the experimentl perio using the following formuls: 1668

3 Worl Appl. Si. J., 15 (12): , 2011 BWG = Finl Boy Weight - Initil Boy Weight prffin, setione t 4-6 mirons thikness n stine Lipi Profile n Lipoprotein Cholesterol Assy: Totl with Hemtoxylin n Eosin stin for exmintion of the lipi (TL) onentrtions were etermine olorimetri liver s esrie y Crleton [23]. using spetrophotometer pprtus just t 520 nm s esrie y kit instrutions (Rnox Co. Ireln). Sttistil Anlysis: Dt were expresse s men ± Triglyeries (TG), totl holesterol (TC) n high ensity stnr error. In orer to ompre the groups, nlysis lipoprotein holesterol (HDL-C) onentrtions were of vrine (ANOVA) ws use. P < 0.05 vlues were etermine using enzymti methos s esrie in the onsiere to e sttistilly signifint. instrutions provie with the kits (Anlytion Biotehnologies AG, Germny). The sorne of the RESULTS testes smples were re using spetrophotometer juste t 546 nm for TG n TC n 500 nm for HDL-C. Foo Intke n Boy Weight: Effets of high ft, Then rtio of high ensity lipoprotein holesterol to totl holesterol-iet either only or supplemente with ifferent holesterol rtio ws mesure. levels of pine nuts on foo intke n oy weight gin Low ensity lipoprotein holesterol (LDL-C) in rts re shown in tle 1. Results revele tht high ft, onentrtion ws lulte y using formul of holesterol-iet signifintly reue foo intke Friewl et l. [22] n very low ensity lipoprotein (18.43±0.48 g/) t p<0.05 s ompre to free holesterolholesterol (VLDL-C) ws lulte using the following iet (23.43 ± 0.72 g/). In ition, high ft, holesteroleqution: iets supplemente with 5, 10 n 15% pine nuts signifintly (p<0.05) reue foo intke (16.14 ± 0.34, LDL-Cholesterol = Totl holesterol - (HDL-C+ TG/5) ± 0.42 n13.14 ± 0.40 g/, respetively) s VLDL- (mg/ L) = TG/5 ompre to high ft, holesterol-iets (18.43 ± 0.48 g/). Men vlues of initil, finl oy weight for ll nimls Liver Funtions Assy: Serum sprtte minotrnsferse were prtilly similr for ll groups n not signifintly (AST) n lnine minotrnsferse (ALT) n lkline hnge. Boy weight gin of positive ontrol group n phosphtse (ALP) tivities were etermine using trete groups with pine nut were no signifint eferene olorimetri methos s esrie in the kits instrution s ompre to the norml ontrol rts. (Dimon Co, Hnnover, Germny). The sorption of the test smples were re t 505nm for GOT n GPT n t Reltive Orgns Weight to Boy Weight: Dt in tle 2 510nm for ALP. reporte tht positive ontrol rts fe high ft, holesterol-iet h signifint inrese in reltive liver, Histopthologil Exmintion: Hert, ort n liver of hert n kineys to oy weight t p<0.05 s ompre the srifie rts were tken n immerse in 10% formlin to negtive ontrol rts fe sl iet. Groups fe high ft, solution. The fixe speimens were then trimme, wshe holesterol-iets supplemente with ifferent levels of n ehyrte in sening gres of lohol. pine nuts h no signifint hnges in reltive liver Speimens were then lere in xylol, emee in weight to oy weight s ompre to the positive Tle 1: Foo intke, oy weight in rts. Prmeters s Men ± SE Groups Foo intke (g/) Initil weight (g) Finl weight (g) Boy weight gin (g) Negtive group 23.43± ± ± ±0.97 Positive group 18.43± ± ± ±0.80 5% Pine nut 16.14± ± ± ± % Pine nut 14.71± ± ± ± % Pine nut e 13.14± ± ± ±0.72 Mens in eh olumn with ifferent supersript letters iffer signifintly t p<0.05. A uses hrmoni men smple size = 7 rts. 1669

4 Worl Appl. Si. J., 15 (12): , 2011 Tle 2: Reltive orgns weight to oy weight in rts. Prmeters s Men ± SE Groups Liver (g) Kineys (g) Hert (g) Negtive group 2.70 ± ± ± 0.01 Positive group 4.26 ± ± ± % Pine nut 4.38 ± ± ± % Pine nut 4.32 ± ± ± % Pine nut 4.64 ± ± ± 0.01 Mens in eh olumn with ifferent supersript letters iffer signifintly t P <0.05. A uses hrmoni men smple size = 7 rts. Tle 3: Conentrtions of serum totl lipi, triglyeries n totl holesterol in rts. Prmeters s Men ± SE Groups Totl lipi (mg/l) Triglyeries (mg/l) Totl holesterol (mg/l) Negtive group ± ± ±.53 Positive group ± ± ± % Pine nut ± ± ±.80 10% Pine nut ± ± ± % Pine nut ± ± ± 2.14 Mens in eh olumn with ifferent supersript letters iffer signifintly t P <0.05. A uses hrmoni men smple size = 7 rts. Tle 4: Conentrtions of serum LDL-C, HDL-C n VLDL-C well s HDL-C/TC rtio in rts. Prmeters s Men ± SE Groups LDL-C (mg/l) HDL-C (mg/l) VLDL-C (mg/l) HDL-C/TC rtio Negtive group 8.57 ± ± ± ± 0.02 Positive group ± ± ± ± % Pine nut ± ± ± ± % Pine nut ± ± ± ± % Pine nut ± ± ± ± 0.02 Mens in eh olumn with ifferent supersript letters iffer signifintly t P <0.05. A uses hrmoni men smple size = 7 rts. Tle 5: Conentrtions of serum of AST, ALT n ALP in rts. Prmeters s Men± SE Groups AST (U/L) ALT (U/L) ALP (U/L) Negtive group 14.27± ± ± 0.35 Positive group ± ± ± % Pine nut ± ± ± % Pine nut ± ± ± % Pine nut ± ± ± 0.26 Mens in eh olumn with ifferent supersript letters iffer signifintly t P <0.05. A uses hrmoni men smple size = 7 rts ontrol group. Reltive kineys weight to oy weight the positive ontrol group (0.61 ± 0.02g). Wheres, high of groups fe high ft, holesterol-iet supplemente levels of pine nuts (15%) signifintly (p<0.05) reue with 5 n 10% pine nuts were not signifint hnge reltive kineys weight to oy weight (0.54 ± 0.02g) s (0.58 ± 0.01 n 0.56 ± 0.02 g, respetively) s ompre to ompre to the positive ontrol group (0.61 ± 0.02g). 1670

5 Worl Appl. Si. J., 15 (12): , 2011 Aministrtion of pine nuts t 5 n 15% use no serum level of LDL-C n VLDL-C mg/l n signifint signifint erese in reltive hert weight to oy erese in serum levels of HDL-C, (p<0.05) ompre to weight, while its level t 10 % use signifint erese negtive ontrol group. Men ± SE vlue of HDL-C/TC s ompre to the positive ontrol group. rtio signifintly reue (0.15 ± 0.09) in positive ontrol group s ompre to the norml ontrol group (0.40 ± Serum Conentrtions of Totl Lipi, Triglyeries 0.02). Rts fe high ft, holesterol-iets supplemente n Totl Cholesterol: Tle 3 shows signifint with ifferent levels of pine nuts h signifint reue inrese (p<0.05) in serum onentrtions of totl lipi (p<0.05) in serum levels of LDL-C n VLDL-C mg/l s (TL), triglyeries (TG) n totl holesterol (TC) in ompre to positive rts. Low level of pine nuts (5%) positive ontrol group ( ± 1.85, ± 2.20 n proue no signifint hnge in serum level of HDL-C, ± 3.01 mg/l, respetively) s ompre to the while 10 n 15% pine nuts use signifint inrese, negtive ontrol group ( ± 1.17, ± 2.24 n ompre to positive ontrol group. Men ± SE vlues for ± 0.53 mg/l, respetively). Rts fe high ft, HDL-C/TC rtio were signifintly inrese (p<0.05) in holesterol-iets supplemente with 5, 10 n 15% pine trete group with ifferent levels of pine nuts s nuts h lower men ± SE vlues for serum ompre to positive group. onentrtions of TL, TG n TC, whih were erese signifintly t p<0.05, exept trete group with 5% h Serum Conentrtions of AST, ALT n ALP: Tle 5 no signifint hnge in serum level of TG s ompre shows signifintly (p<0.05) higher serum AST, ALT n the positive ontrol group. ALP (25.21 ± 0.50, ± 0.63 n ± 0.67 U/L, respetively)levels in positive ontrol rts s ompre to Serum Conentrtions of LDL-C, HDL-C, VLDL-C s the negtive ontrol rts (14.27± 0.44, ± 0.48 n Well s HDL-C /TC Rtio: The effet of high ft, ± 0.35 U/L, respetively). Aministrtion of ifferent holesterol-iet on serum onentrtions of LDL-C, levels of pine nuts resulte in signifint reution in VLDL-C n HDL-C re shown in tle 4. Results revele the level of serum AST, ALT n ALP (p<0.05) s tht positive ontrol group h signifint inrese in ompre to positive ontrol group Fig. 1: Hert of rts from negtive ontrol group showing no histopthologil hnges (H n E x 400). Fig. 2: Hert of rts from positive ontrol group showing ongestion of ri loo vessel n hylinosis of its wll (H n E x 400). Fig. 3: Hert of rts from positive ontrol group showing grnulrity of the sroplsm of fol ri myoytes (H n E x 400). Fig. 4: Hert of rts from trete group with 5% pine nuts showing fol nerosis of myoytes ssoite with leuoyti ells infiltrtion n ongestion myoril loo vessels (H n E x 400). 1671

6 Worl Appl. Si. J., 15 (12): , Fig. 5: Aort of rts from negtive ontrol group showing no histopthologil hnges (H n E x 400). Fig. 6: Aort of rts from positive ontrol group showing prolifertion of smooth musles of tuni mei ssoite with thikening (H n E x 200). Fig. 7: Aort of rts from positive ontrol group showing vultions of tuni mei (H n E x 400). Fig. 8: Aort of rts from trete group with 5 n 10% pine nuts showing vultions of tuni mei (H n E x 200) Fig. 9: Liver of negtive ontrol rts showing the norml histologil struture of hepti loulr (H n E x 400). Fig. 10: Liver of positive ontrol rts showing ftty hnge of heptoytes (H n E x 400). Fig. 11: Liver of trete rts with 5, 10 n 15% pine nuts showing ftty hnge of heptoytes (H n E x 400). Histopthologil Exmintion were showe in exmine hert setions of rts fe high Hert: Hert setions of norml rts show norml ft, holesterol-iet supplemente with 5% pine nut histologil struture s shown in figure 1. In ontrst, (Figure 4). Wheres, hert setions of rts fe high ft, hert setions of rts fe high ft, holesterol-iet holesterol-iet supplemente with 10 n 15% pine nuts revele ongestion of ri loo vessel n showe norml histologil struture. hylinosis of its wll (Figure2), in ition to grnulrity of the sroplsm of fol ri myoytes (Figure 3). Aort: Histopthologil exmintion of ort setions of Fol nerosis of myoytes ssoite with leuoyti negtive ontrol rts revele no histopthologil ells infiltrtion n ongestion myoril loo vessels hnges s shown in figure 5. However, prolifertion of 1672

7 Worl Appl. Si. J., 15 (12): , 2011 smooth musles of tuni mei ssoite with The present results showe tht positive ontrol thikening were oserve in ort of rts from positive group h signifint inrese in the serum level of totl ontrol group fe high ft, holesterol-iet (Figure 6), in lipi (TL), triglyeries (TG) n totl holesterol (TC), ition to vultions of tuni mei (Figure 7). Aort LDL-C, VLDL-C, however, signifint erese in serum setions of rts fe high ft, holesterol-iet HDL- C, ompre to the negtive ontrol group. The supplemente with 5 n 10% pine nuts h vultions inreses in serum onentrtions of the ove mentione of tuni mei (Figure 8). However, rts fe iet prmeters n the reution in serum HDL-C s results supplemente with 15% pine nuts show norml of high ft, holesterol iet hve een pointe out s risk histologil struture. ftors for the evelopment of theroslerosis n relte riovsulr iseses, whih ws represente y the Liver: Histopthologil exmintion of livers from erese in HDL-C/TC rtio. These results were onfirme negtive ontrol rts revele norml histologil y histopthologil exmintion of hert whih revele struture of hepti loule s shown in Figure 9. Liver ongestion of ri loo vessel n hylinosis of its setions of positive rts h ytoplsmi wll n grnulrity of the sroplsm of fol ri vuoliztion of heptoytes n ftty hnges of myoytes. In ition to, ort setions h prolifertion heptoytes (Figure 10). Ftty hnges of heptoytes of smooth musles of tuni mei ssoite with were showe in exmine liver setions of ll groups fe thikening n vultions of tuni mei exmine iet supplemente with 5 n 10 n 15% pine nuts ort. The present results gree with Gregorio et l. [26] (Figure 11). who showe tht serum triglyeries, holesterol, LDL-C, VLDL-C inrese signifintly, however, serum HDL-C DISCUSSION erese signifintly in rts fe high holesterol iet, ompre to ontrol rts. In ition to, Cohn [27] The present stuy ws rrie out to investigte the revele tht hyperlipiemi in rts is hrterize y the hypolipieimi n hypoholestermi effet of pine nuts rise in triglyerie-rih lipoproteins fter rih ft mel t ifferent levels on rts fe high ft, holesterol iet. n it is ssoite with n inrese risk of oronry The present results revele tht high ft, rtery isese. Triglyerie-rih lipoproteins n penetrte holesterol-iet signifintly reue foo intke, the rtery wll n my hve iret effet on ompre to norml sl iet. These results my e theroslerosis. Zhen-Yu n Xio-Qin [28] ttriute to the higher lori ontent of high ft, emonstrte tht lipi metolism in rts fee high ft holesterol iet s ompre to norml sl iet. The iet presente isorer n the level of serum TC n inrese in ft ontent of the iet re responsile for triglyeries inrese signifintly, ompre with those stiety n inrese totl lories. This result gree in the norml ontrol group. with Sheyl et l. [24] who emonstrte tht high ft Clinil stuies inite tht hyperholesterolemi ontent iet, whih is use to inue is n essentil risk ftor for oronry rtery isese hyperholesterolemi, les to lower ingestion y the (CAD), where low-ensity lipoprotein (LDL) holesterol nimls n inues mlnutrition. plys mjor role in the theroslerosis n pthogenesis In reltion to reltive liver n hert weight to of CAD n other vsulr iseses [29]. These results oy weight, there ws signifint inrese in onfirme y Kumr et l. [30] who reporte tht rts fe hyperholesterolemi group s ompre to norml group. high ft iet h signifint elevtion in plsm totl n These results might e expline se on the LDL-holesterol n triglyerie n the rtio of HDLumultion of ft in the liver n hert ells. These C/totl holesterol ws signifint erese in positive results were onfirme y histopthologil exmintion group, ompre the norml ontrol group. whih showe tht ftty hnges of heptoytes n The pprent results might e expline y the grnulrity of the sroplsm of fol ri myoytes. tivity of lipoprotein lipse s reporte y Tei et l. These results were in orne with Sheyl et l. [24] [31] who foun tht tivity of this enzyme ugmente in who reporte tht the inrese in liver weight oul e hyperholesterolemi nimls. Lipse trnsforms VLDL onsequene of their higher ft ontent (ft/liver). In to LDL-holesterol, wht woul le to n inrese in ition to Pusks et l. [25] showe tht intrellulr the serum onentrtion of LDL-C. On the other hn lipi umultion in riomyoytes in response to Berg et l. [32] emonstrte tht uptke of LDLholesterol iet. holesterol is epenent on reeptors in plsmti 1673

8 Worl Appl. Si. J., 15 (12): , 2011 memrne n these re in reue numer when The present results were noteworthy n might e the ell hs enough holesterol. This my hve ue to the type of ftty is omposition in pine nuts hppene in hepti ells of the niml fe holesterol- espeilly pinoleni i whih is the min ftty is in supplemente iets, explntory their higher LDL- pine see oil, or to some of its metolites, euse this holesterol onentrtion. unusul ftty i is the min ifferene in the ft Assmnn et l. [33] n Yrnell et l. [34] omposition of the two experimentl iets. These results reporte tht hypertriglyeriemi inrese the risk gree with Sugno et l. [39] n Mtsuo et l. [40] who of ute oronry events n some linil trils reporte tht pine nuts oil i not influene the growth foun high serum triglyeries to e n inepenent performne of trete nimls in omprison to other risk ftor for riovsulr isese. Furthermore, ietry fts. Asset et l. [6] foun tht oy weight, hyperholesterolemi omine with mrke foo intke n liver weight were not signifintly hypertriglyeriemi les to ysfuntion in hert of ifferent etween rts trete with pine nut oil n their rts [35]. respetive ontrols. These results were onfirme with Severl stuies showe tht hyperlipiemi inues Yiliz-Guly et l. [41] who reporte tht there ws no oxitive stress n this oxitive moifition of hnge of oy weight gin n verge foo intke in lipoproteins in vessel wlls ply key role in rits fee iet supplemente with pine nuts. The therogenesis [36]. Hyperlipiemi exerts omplex effets weights of the orgns were similr n there were no on the myorium. Intrellulr lipi umultion in signifint ifferenes in orgn weights etween ontrol riomyoytes n severl ltertions in the struturl n tretment groups. Wheres, Ferrmos et l. [42] n funtionl properties of the myorium hve een showe tht pine nuts oil use signifint reution oserve in response to holesterol iet [25, 37]. in oy weight gin n liver weight. In ition to, Results lso, reore tht positive ontrol rts h Zhen-Yu n Xio-Qin, [28] revele tht the inrese signifint elevtion in serum level of AST, ALT n ALP, in oy weight of trete rts with pine nuts oil ws muh ompre to the norml ontrol rts. The elevtions in the less thn tht of the rts in high ft iet group. tivity of liver enzymes my e ttriute to their relese Furthermore, stuies rrie out in humns hve from the ytoplsm into the loo irultion fter rupture emonstrte tht pine nuts stimultes the relese of the of the plsm memrne, ellulr mge. These results stiety gut hormones holeystokinin (CCK) n onfirme y histopthologil stuy, whih oserve glugon like peptie-1 (GLP-1), therey leing to tht liver setions of h ytoplsmi vuoliztion n reue fee intke. The suggestion tht pine nuts oul ftty hnges of heptoytes. These results gree with e use s n ppetite suppressnt in humns hs Tei et l. [31] who showe tht the tivity of this therefore emerge [43]. enzyme inrese in hyperholesterolemi nimls. The The mjor fining of the present stuy ws the uptke of LDL-holesterol is epenent on reeptors in reution in serum levels of totl lipi, triglyeries, totl plsmti memrne. This my e hppene in hepti holesterol, LDL-C, VLDL-C, AST, ALT n ALP, whih ells of the niml fe holesterol-supplemente iets, were signifint erese n signifint elevtion in explntory their higher LDL-holesterol onentrtion. serum level HDL-C in trete rts, exept low level of pine Pinus n Shffner [38] reporte tht this effet might nuts (5%) inue non signifint hnge in TG n HDLe relte to pproximtely 80% of AST in heptoytes C, ompre to the positive ontrol rts. It revele tht ppers to e lote in mitohonri, wheres ALT is the lowest reution of the ove serum prmeters were thought to e preominntly no mitohonril n it hs etete with inreses the e level of pine nuts into een postulte tht in mil heptoellulr injury. When high ft, holesterol iet. Therefore, pine nuts my e the heptoytes plsmti ut not the mitohonril erese the iniene of theroslerosis n oronry memrne is mge, ytoplsmti AST n ALT re hert ieses, whih ws more etetle with the inrese relese into serum with more severe heptoellulr injury in serum level of HDL-C n HDL-C/TC rtio. These n mitohonril memrne mge. results were onfirme with the otine results of With regr to the effet of pine nuts in rts fe high histopthologil exmintion. ft, holesterol iet, results revele tht ifferent levels The present results gree with Sugno et l. [39] of pine nuts reue signifintly foo intke s ompre n Asset et l. [44] who showe tht pine nuts to positive rts. It lso reue men vlues of oy supplementtion lowers loo lipis n ffets weight gin n reltive liver, hert n kineys weight to lipoprotein metolism. Reent stuies inite tht oy weight, ompre to the positive ontrol group. ition of pine nuts to the iet tene to erese 1674

9 Worl Appl. Si. J., 15 (12): , 2011 plsm holesterol onentrtions [41] n use no 3. Megre, V., The Ringing Cers Series, ook 1. signifint ltertions on serum ALT, AST n ALP levels Columi, Missouri: Ringing Cers Press, pp: 224. for rits [45]. 4. Brufu, G. n J. Botell, Rfes Nuts: Soure The possile mehnism effet of pine nuts my e of energy n mronutririons. Br. J. Nutr., 96: relte to its ftty i omposition whih is n importnt 5. Ste, J., Nut onsumption, vegetrin iets, ftor ple of moulting lipi metolism. These ishemi hert isese risk n ll uses mortlity: results gree with Asset et l. [6] 7ho showe tht levels Eviene from epiemiologi stuies. Am. J. Clin. of serum triglyeries were erese signifintly in the Nutr., 70: group supplemente with pine nuts oil ompre to its 6. Asset, G., B. Stels, R.L. Wolff, E. Buge, Z. Mj, respetive ontrol group. This reution ws ounte J.C. Fruhrt n J. Dllongeville Effets of for y erese in VLDL-holesterol. Pinus pinuster n Pinus koriensis see oil In ition, it must e onsiere tht other supplementtion on lipoprotein metolism in the rt. iotive ompouns present in the pine nuts iet my Lipis, 34: hve role in triggering the effets oserve in this 7. Lwlees, J., The enylopei of essentil oils. stuy. These results gree with Woo et l. [46] who Element Books Limite: Boston, pp: mentione tht pine nuts extrt re rih in 8. Gr, J. n A. Lis, Olejki sosnowe. pronthoyniins n ontin rnge of flvonois Aromterpi, 2: inluing tehin, epitehin, queretin, 9. Sgrero-Nieves, L., Ftty i omposition of ihyroqueretin, txifolin n phenoli is. Therefore, Mexin pine nut (Pinus emroies) oil from three the improvement effets of pine nuts on lipi profile n see ot phenotypes, J. Siene of Foo n lipoprotein holesterol might e relte to its ntioxint Agriu. 59: tion. Previous reserh inite tht riovsulr 10. Svge, G.P., Chemil omposition of wllnuts risk ftors re ssoite with ognitive eline [47]. (Juglns regi L.) grown in New Zeln. Pln Foo However, ioflvonoi onsumption improve Hum. Nutr., 56: riovsulr risk ftors n promotes relxtion of 11. Forester, J.S., Triglyeries: risk ftor or fellow vsulr smooth musle [48] n improvements of loo trveler? Curr Opin Criol., 16: irultion to etter ognitive funtions [49]. Previous 12. Huek, J.A., V. Amkov, M. Vrlik, M. stuies hve shown tht the potentil effet of pine nuts Kleov, J.Zih, J. Kunes, J. Pith, P. Suhnek for riovsulr enefits inlues improvement in n R. Polene Apolipoprotein A5 in helth enothelil funtion, reution in plsm firinogen n isese. Physiol. Res., 58: onentrtions [50], reue plsm visosity n 13. Gerhrt, A.L. n N.B. Gllo, Full-ft rie rn systoli loo pressure [51]. n ot rn similrly reue hyperholesterolemi in humns. J. Nutr., 128: CONCLUSION 14. Roins, S.L., Ptologi e struturl funtionl. Rio e Jneiro: Gunr-Koogn, pp: The present results revele tht the ifferent levels 15. Gomes, M.R., H. Tomso, G.K. Nzrin, of pine nuts use reution in foo intke, oy weight H. Bjrnson, C.A. Dietz n D.W. Hunter, gin n reue the iniene of hyperlipiemi n Upper extremity eep vein thromosis n hyperholestermi in rts fe high ft, holesterol iet s hroni pulmonry emolism resulting in pulmonry well s liver enzymes onentrtions. The lowest rtery hypertension. AJR Am. J. Roentgenol., reution ws more etetle with inrese the level of 170: pine nuts. 16. Mhn L.K. n S. Sott, Kruse - Alimentos, Nutrio Dietoterpi. So Pulo: ROCA. REFERENCES 17. Ferinny, P., Z. Szilvssy n G.F. Bxter, Apttion to myoril stress in isese sttes: is 1. Lnner, R., The Pinon pine. A Nturl n preonitioning helthy hert phenomenon? Culturl History. University of Nev Press, Trens. Phrmol. Si., 19: pp: Ferinny, P., Myoril ishemi/ 2. Ozguven, F. n K. Vursvus, Some physil reperfusion injury n preonitioning: effets of mehnil n eroynmi properties of pine hyperholesterolemi/hyperlipiemi. Br J. (pinus pine) nuts. J. Foo Eng., 68: Phrmol., 138:

10 Worl Appl. Si. J., 15 (12): , Chttophyy, R., D. Pthk n D.P. Jinl, 33. Assmnn, G., H. Shulte n A. Von Ekrstein, Antihyperlipiemi gents A review. Inin Drugs, 33: Reeves, P.G., F.H. Nielson n G.C. Fhmy, Reports of the Amerin Institute of Nutrition, ho-wiling ommittee on reformultion of the AIN 93, Roent iet. J. Nutr., 123: Blkn, J., S. Dogru-Asoglu, G. Ayk-Toker n Hypertriglyeriemi n elevte lipoprotein () re risk ftors for mjor oronry events in mile-ge men. Am. J. Criol., 77: Yrnell, J.W., C.C. Ptterson, P.M. Sweetnm, H.F. Thoms, D. Binton n P.C. Elwoo, Do totl n high ensity lipoprotein holesterol n triglyeries t inepenently in the preition of M. Uysl, The effet of high holesterol iet on lipis n oxitive stress in plsm liver n ort of rits n rts. Nutr. Res., 24: Friewl, W.T., R.I. Levy n D.S. Frerikson, Estimtion of the onentrtion of low-ensity lipoprotein holesterol in plsm, without use of the preprtive ultrentrifuge. Clin Chem., 18: ishemi hert isese? Ten-yer follow-up of th 23. Crleton, H., Histologil tehniques. 4 E, Lonon, Oxfor University Press, New York, USA, Toronto, pp: Sheyl, L.M., H. De Pul1, L.P. Mri, C. Rinlo, E. Sntos, L. Deoléio n E.S. Mrelo, Dietry moels for inuing hyperholesterolemi in rts. Brzilin Arhives of Biology n Teh., 48: Pusks, L.G., Z.B. Ngy, Z. Giriz, A. Onoy, C. Csonk, K. Kitjk, L. Hkler, A. Zvr n P. Ferinny, Cholesterol iet-inue hyperlipiemi influenes gene expression pttern of rt herts: DNA mirorry stuy, FEBS Lett., 562: Gregorio, S.R., M.A. Ares n F.G. Reyes, Dietry fiers n riovsulr isese. Nutrition, 22: Cohn, J.S., Postprnil lipemi n remnnt lipoproteins. Clin L. Me. 26: Zhen-Yu, W. n C. Xio-Qin, Funtionl evlution for effetive ompositions in see oil of Koren pine, J. Forestry Res., 15: Truelj, N., C. Vughn n N.L. Copln, The role of sttins in preventing stroke. Preventive Criol., 8: Kumr, D.S., A.K. Muthu, A.A. Smith n R. Mnvln, Hypolipiemi effet of vrious extrts of whole plnt of Muun pruriens (Linn) in rt fe with high ft iet. Eur. J. Biol. Si., 2: Tei, K., J.M. Rounet n P. Besnon, Effet of grpe see tnnins on the tivity of some rt intestinl enzyme tivities. Enzyme Protein, 48: Berg, J.M., J. Tymozko n L. Stryer, Biohemistry. 5th e. New York : Freemn n Compny. Cerphilly n Speewell Cohorts. Arteriosler Throm Vs Biol., 21: Onoy, A., C. Csonk, Z. Giriz n P. Ferinny Hyperlipiemi inue y holesterol-rih iet les to enhne peroxynitrite formtion in rt herts. Criovs Res. 58: Wittenstein, B., M. Klein, B. Finkh, K. Ullrih n A. Kohlshutter, Plsm ntioxints in peitri ptients with glyogen storge isese, ietes mellitus n hyperholesterolemi. Free Ril Biology n Me., 33: Hexeerg, S., N. Willumsen, S. Rotevtn, E. Hexeerg n R.K. Berge, Cholesterol inue lipi umultion in myoril ells of rts. Criovs Res., 27: Pinus, M.R. n J.A. Shffner, Assessment of liver funtion. In: Henry JB. Clinil ignosis n mngement y lortory methos. Philelphi, W.B. Suners Compny, pp: Sugno, M., I. Ike, K. Wkmtsu n T. Ok, Influene of Koren pine (Pinus koriensis)- see oil ontining is-5,is-9,is-12-otetrienoi i on polyunsturte ftty i metolism, eiosnoi proution n loo pressure of rts. Brit. J. Nutr., 72: Mtsuo, N., K. Os, T. Kom, B.O. Lim, A. Nko, K. Ym n M. Sugno, Effets of gmm-linoleni i n its positionl isomer pinoleni i on immune prmeters of rown- Norwy rts. Prostgln Leuk Essent Ftty, 55: Yiliz-Guly, O., M.S. Guly, A. At, A. Bli n A. Demirts, The effets of feeing pinus pine sees on some loo vlues in mle New Zeln white rits. J. Anim n Veteri Avn., 9: Ferrmos, A., V. Svy, L. Conte, S. Colomo, A.W. Einerhn, V. Zr, Conjugte linolei i n hepti lipogenesis in mouse: role of the mitohonril itrte rrier. J. Lipi Res., 47:

11 Worl Appl. Si. J., 15 (12): , Hughes, G.M., E.J. Boyln, N.J. Willims, 48. Cett, R., V. Burke, T. Mori, L. Beilin, L. Puey L. Mennen, C. Sott, T.C. Kirkhm, J.A. Hrrol, n K. Crft, Re wine polyphenols in H.G. Keizer n J.C. Hlfor, The effet of the sene of lohol reue lipi peroxitive Koren pine nut oil (PinnoThin) on foo intke, stress in smoking sujets. Free Ri. Biol. Me., feeing ehvior n ppetite: oule-lin 30: pleo-ontrolle tril. Lipis Helth Dis, pp: Stein, J., J. Keevil n S. Wiee, Purple grpe 44. Asset, G., E. Buge, R.L. Wolff, J.C. Fruhrt n juie improves enothelil funtion n reues J. Dllongeville, Effets of ietry mritime pine suseptiility of LDL holesterol to oxition in see oil on lipoprotein metolism n ptients with oronry rtery isese. Cirultion, theroslerosis evelopment in mie expressing 100: humn polipoprotein B. Eur. J. Nutr., 40: Young, J.M., B.I. Shn, P.M. MGregor, R.S. Sott 45. Hein, J. n J. Hrtmnn, Lorignostishe n C.M. Frmpton, Comprtive effets of Referenz-ereihe ei Kninhen. Tiererztl Prx, enzogenol n vitmin C supplementtion versus 31: vitmin C lone on enothelil funtion n 46. Woo, J.E., S.T. Senthilmohn n A.V. Peskin, iohemil mrkers of oxitive stress n Antioxint tivity of proyniin-ontining plnt inflmmtion in hroni smokers. Free Ri. extrts t ifferent phs. Foo Chem., 77: Res. 40: Brnes, D.E., G.S. Alexopoulos, O.L. Lopez, 51. Shn, B., C. Strey, R. Sott, Z. Morrison n J.D. Willimson n K. Yffe, Depressive S. Gieseg, Pilot stuy on the linil effets of symptoms, vsulr isese n mil ognitive ietry supplementtion with Enzogenol, flvonoi impirment: finings from the Criovsulr Helth extrt of pine rk n vitmin C. Phytother Stuy. Arh. Gen. Psyhitry, 63: Res., 17: