Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 RESEARCH ARTICLE Open Access Effects of red pity juice supplementtion on crdiovsculr nd heptic chnges in high-crohydrte, high-ft diet-induced metolic syndrome rts Nurul Shzini Rmli 1, Lindsy Brown 2, Ptimh Ismil 3 nd Asmh Rhmt 1* Astrct Bckground: The fruit of Hylocereus polyrhizus, lso known s red pity, nd uh ng in Mly, is one of the tropicl fruits of the cctus fmily, Cctcee. Red pity hs een shown to protect ort from oxidtive dmge nd improve lipid profiles in hypercholesterolemic rts proly due to phytochemicls content including phenolics nd flvonoids. The im of this study ws to investigte the chnges in crdic stiffness, heptic nd renl function in high-crohydrte, high-ft diet-induced oese rts following supplementtion of red pity juice. Methods: Totl 48 mle Wistr rts were divided into 4 groups: corn-strch group (CS), corn-strch + red pity juice group (CRP), high-crohydrte, high ft group (HCHF) nd high-crohydrte, high ft + red pity juice (HRP). The intervention with 5% red pity juice ws strted for 8 weeks fter 8 weeks initition of the diet. Hert function ws determined ex vivo with Lngendorff herts while plsm liver enzymes, uric cid nd ure were mesured using commercil kits. Totl ft mss ws determined with Dul-energy X-ry sorptiometry (DXA) scn. Glucose uptke ws mesured with Orl Glucose Tolernce Test (OGTT). Liver nd crdic structures were defined y histology. Results: Supplementtion of red pity juice for 8 weeks incresed energy intke nd dominl circumference ut no chnge in ody ft nd len mss respectively. Also, there were trend of uric cid nd glucose normliztion for HRP s compred to H-fed rts. Red pity juice tretment reduced ALP nd ALT ut cused significnt increment in AST. Distolic stiffness of the hert ws reduced fter supplementtion of red pity juice in corn strch fed rts. However, the reduction ws not significnt in HRP rts in comprison with H rts. Conclusion: The present study concluded tht red pity juice my serve s complimentry therpy for ttenuting some signs of metolic syndrome. Keywords: Red pity juice, Metolic syndrome, High-crohydrte high-ft diet Bckground Overweight nd oesity re drmticlly on the rise in recent decdes. According to WHO [1], oesity contriuted to doule urden of diseses prticulrly dietes (44%), ischemic hert diseses (23%), nd certin types of cncer (7-41%). This is due to the metolic normlities creted y excessive ft ccumultion like normlities of * Correspondence: smh@upm.edu.my 1 Deprtment of Nutrition nd Dietetics, Universiti Putr Mlysi, Serdng 43400 UPM, Mlysi Full list of uthor informtion is ville t the end of the rticle lipid in the lood, hypertension nd impired glucose tolernce, mong which re the common fetures of metolic syndrome [2]. In ptients with metolic syndrome, insulin resistnce results in the impired insulin ctivities in tissues like muscle, liver, kidney nd ft leding to increse oxidtive stress, pro-cogulnt/nti-firinolytic nd chronic pro-inflmmtory stte coupled with pltelet hyper-ggreglity [3]. Aville evidences suggested the use of dietry intervention s n integrl prt of future pproches to prevent nd tret oesity nd its metolic consequences [4]. Hence, this study focuses on 2014 Rmli et l.; licensee BioMed Centrl Ltd. This is n Open Access rticle distriuted under the terms of the Cretive Commons Attriution License (http://cretivecommons.org/licenses/y/2.0), which permits unrestricted use, distriution, nd reproduction in ny medium, provided the originl work is properly credited. The Cretive Commons Pulic Domin Dediction wiver (http://cretivecommons.org/pulicdomin/zero/1.0/) pplies to the dt mde ville in this rticle, unless otherwise stted.
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 2 of 10 crdiovsculr nd heptic system s they re the ultimte consequences of oesity. Diet-induced metolic syndrome ws found to e the closest model tht t lest shres the similr ethologic, nd hence more representtive of humn pthophysiology of metolic syndrome. Humn consume high mount ft nd crohydrte like sucrose nd fructose in their diet. Pnchl et l. [5] studied the remodelling effect of highcrohydrte high-ft diet-induced oesity in rts using condensed milk (39.5%), eef tllow (20%), nd fructose (17.5%) with 25% fructose in drinking wter nd found tht rts developed crdiovsculr, metolic, renl, heptic nd pncretic chnges. The complictions includes oesity, incresed ft ccumultion in dominl region, hypertension, insulin resistnt, nd impired crdic function, endothelil dysfunction s well s inflmmtion. Hence, it cn e seen tht comintion of high crohydrte nd high ft diet produce more humn-like model. This study only utilized mle rts to void the influence of the oestrus cycle on food intke which my ffect the dietinduced model [6]. Consumption of fruits nd vegetles hs long een linked to the prevention of oxidtive stress relted diseses like dietes mellitus, cncer, hert disese, oesity nd micronutrient deficiencies [7-10]. Eting fruits nd vegetles cn ensure the dequte supply of micronutrients, dietry fiers nd phytochemicls which in turn mintin the ody in helthy stte [11]. However, it is not cler which specific fruits nd vegetles re most protective ginst certin diseses. Only few studies hve exmined the effect of specific fruits or juices on metolic syndrome risk fctors. A lrge prospective cohort study for 10.2 yers on Swedish men nd women found significntly inverse ssocition of only pples, pers nd green lefy vegetles with stroke [12]. Not ll fruits re creted equl prticulrly in terms of their phytonutrient contents which might influence their iologicl properties, nd hence their efficcy in reltion to specific diseses. The fruit of Hylocereus polyrhizus, lso known s red pity, nd uh ng in Mly, is one of the tropicl fruits of the cctus fmily, Cctcee. Polyphenols including flvonoids, etcynins, vitmin C nd fier re mong the min ctive constituents in red pity known to confer helth enefit [13-15]. However, etcynin frctions shown to disply the highest reducing nd rdicl scvenging cpcities s compred to polyphenolic frctions [16]. Since the study on the physiologicl effects of red pity is still some distnce from tht of other fruits, it is interesting to investigte whether supplementtion of 5% red pity juice cn meliorte the metolic, heptic nd renl function in rts fed highcrohydrte, high-ft diet. To the est of our knowledge, this is the first study to evlute the effect of red pity ssocited with heptoprotection nd crdioprotection. Methods Preprtion of diet Red pity ws otined from Queenslnd Austrli. The identifiction of the fruit ws done y otnist from Biodiversity Unit, Institute of Biosciences, Universiti Putr Mlysi. The voucher numer is SK-2440/14. The fruits were then clened, nd the fruit pulp ws squeezed using juice mker. Smple preprtion ws conducted in reduced light condition in order to minimize the pigment loss. Animls nd diet All experimentl protocols were pproved y the Animl Experimenttion Ethics Committee of The University of Southern Queenslnd under the guidelines of the Ntionl Helth nd Medicl Reserch Council of Austrli. This study ws rndomized control tril. The experimentl groups consisted of 48 mle Wistr rts (ged 8 9 weeks; weight 337 ± 5 g) supplied y nd individully housed t The University of Southern Queenslnd niml house. All experimentl groups were housed in temperturecontrolled, 12 hour light drk cycle environment with d liitum ccess to wter nd food. Dily ody weight, feed nd wter mesurements were tken to monitor the dyto-dy helth of the rts. The rts were rndomly divided into six groups sed on their diet: corn strch (C; n = 12); corn strch + red pity juice (CRP; 5% in the diet; n = 12); high-crohydrte, high-ft (H; n = 12); High-crohydrte, high-ft + red pity juice (HRP; n = 12). Fructose (25%) ws dded s drinking wter for ll high-crohydrte, high-ft fed rts, while corn strch group ws given norml wter. The detiled mcro- nd micro-nutrient composition of the C nd H diets re reported in our previous pulictions [17,18]. Red pity juice supplementtion ws dministered for 8 weeks strting from 8 weeks fter the initition of the C or H diet. Orl glucose tolernce test Bsl lood glucose concentrtions were mesured in til vein lood using Medisense Precision Q.I.D glucose meter (Aott Lortories) fter overnight (10 12 h) food deprivtion. Fructose-supplemented drinking wter in the H nd HRP groups ws replced with norml wter for the overnight food-deprivtion period. The rts were given 2 g/kg ody weight of glucose s 40% solution vi orl gvge. Blood smples from the til vein were tken t 0, 30, 60, 90, nd 120 minute following glucose dministrtion. Body composition mesurements Body composition ws mesured on rts y dul-energy X-ry sorptiometry (DXA) using Norlnd XR36 DXA instrument (Norlnd, Fort Atkinson, WI, USA) fter 16 weeks of feeding nd 2 dys efore terminl
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 3 of 10 experiments. DXA scns were nlysed using the mnufcturer s recommended softwre for use in lortory nimls (Smll Suject Anlysis Softwre, version 2.5.3/1.3.1; Norlnd Corp) [19]. The precision error of len mss for replicte mesurements, with repositioning, ws 3.2%. Viscerl diposity index (%) ws clculted s ([retroperitonel ft (g) + omentl ft (g) + epididyml ft (g)]/[ody weight (g)]) 100 nd expressed s diposity percent [17]. Isolted hert preprtion Lngendorf hert preprtions were used to ssess left ventriculr function of the rts in ll tretment groups. Terminl nesthesi ws induced vi intrperitonel injection of pentoritone sodium (Lethr, 100 mg/kg). Heprin (Sigm-Aldrich Austrli) ws dministered (200 IU) through the right femorl vein nd lood (~5 ml) ws drwn out of the dominl ort. Isovolumetric ventriculr function ws mesured y inserting ltex lloon ctheter into the LV connected to Cpto SP844 MLT844 physiologicl pressure trnsducer nd Chrt softwre on Mcl system (ADInstruments Austrli nd Pcific Islnds). All left ventriculr end-distolic pressure vlues were mesured while pcing the hert t 250 ets/ min using n electricl stimultor. End-distolic pressures were otined strting from 0 mm Hg up to 30 mm Hg. The distolic stiffness constnt (k, dimensionless) ws clculted s in previous studies [20]. +dp/dt nd dp/dt were clculted s the men rte of contrction nd relxtion, respectively, of t lest 50 ets with the hert pced t 250 ets/min, nd the end-distolic pressure ws mintined t pproximtely 10 mmhg. Orgn weights The right ventricle nd LV were seprted fter perfusion experiments nd weighed. Liver nd dominl ft were immeditely removed t the time of the hert removls for perfusion experiments nd lotted dry for weighing. Perirenl, epididyml, nd omentl ft were together weighed s dominl ft. Orgn weights were normlized reltive to the tiil length t the time of their removl (in mg/mm). Plsm iochemistry nlysis Blood ws collected into heprinized tues nd then centrifuged t 5,000 g for 15 minutes. Plsm smples were seprted nd into Eppendorf tues nd stored t 80 C for nlysis. Enzymtic ctivities nd nlyte concentrtions in the plsm (AST, ALP, ALT) were determined using kits nd controls supplied y Olympus using n Olympus nlyzer (AU 400). Plsm glucose, uric cid nd ure were estimted using commercil kit ccording to the mnufcturer-provided stndrds nd protocol using Roche/Hitchi cos c system. Histology The liver nd hert tissues for 2 rts (n = 2) from ech group were exclusively tken for histopthologicl nlysis. The smples were immeditely fixed in 10% formlin for 3 dys to remove the trces of lood from the tissue. After tht, the smples were dehydrted, emedded in prffin wx nd then cut into thin sections (5 6 μm). In order to determine the inflmmtory cell infiltrtion, the liver nd hert tissue sections were stined with hemtoxylin nd eosin. Picrosirius red stining ws used to study collgen deposition in left ventricle of the hert nd ws nlysed using lser confocl microscopy (Zeiss LSM 510 upright confocl microscope). From ech tissue smple, three slides were prepred nd two rndom, nonoverlpping fields were selected from ech slide. A representtive picture ws rndomly selected from ech group. Sttisticl nlysis All dt were presented s men ± SEM. A totl of 4 groups were nlysed using two-wy nlysis of vrince (ANOVA). Ech group consists of 12 rts. All group dt were tested for vrince using Brtlett s test. Vriles tht were not normlly distriuted were trnsformed (using log 10 function) prior to sttisticl nlysis. The effects of diet, tretment nd their interctions were tested y two-wy nlysis of vrince. When interction nd/or the min effects were significnt, mens were compred using Newmn-Keuls multiple-comprison post hoc test. Where trnsformtions did not result in normlity or constnt vrince, Kruskl-Wllis nonprmetric test ws performed. P <0.05 ws considered significnt. All sttisticl nlyses were performed using GrphPd Prism version 5.00 for Windows (Sn Diego, CA, USA). Results Dietry intke The effects of red pity juice on food intke, wter intke nd energy intke were determined fter 8 weeks of supplementtion. Tle 1 shows red pity supplementtion significntly increse food intke only in CRP group. Averge food intke for corn strch fed control group (C) ws 36.33 ± 0.9 g/dy incresed to 40.40 ± 0.7 g/dy for CRP group. On the other hnd, no chnge in food intke ws oserved in high-crohydrte, high-ft diet supplemented with red pity (27.29 ± 1.3 nd 27.74 ± 2.6 g/dy for H nd HRP groups, respectively). As shown in Tle 1, red pity supplementtion did not chnge wter intke in either group (29.57 ± 2.6, 30.88 ± 1.8, 30.15 ± 1.6, 26.47 ± 2.3, 29.56 ± 5.0 nd 21.75 ± 1.1 ml/dy for C, CRP, H, nd HRP groups, respectively). Even though food nd wter intke were similr in high-crohydrte, high-ft diet feeding throughout the study period, rts on this diet supplemented with red pity juice hd incresed in energy
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 4 of 10 Tle 1 Dietry intkes, ody composition, orgn wet weights nd plsm iochemistry nlysis in C, CRP, H nd HRP diet-fed rts Vrile C CRP H HRP P-vlue Diet Tretment Interction Food intke, g/d (n = 7-8) 36.3 ± 0.9 40.4 ± 0.7 27.3 ± 1.3 c 27.7 ± 2.6 c <0.0001 0.0375 0.07 Wter intke, ml/d (n = 7-8) 29.6 ± 2.6 30.9 ± 1.7 26.5 ± 2.3 29.6 ± 5.0 0.06 0.3 0.41 Energy intke, kj/d (n = 8) 405.5 ± 10.2 d 512.7 ± 8.3 c 569.2 ± 28.7 655.2 ± 28.7 0.8041 <0.0001 <0.0001 Body weight gin (8 16 weeks),% (n = 8) 7.1 ± 1.1 c 11.6 ± 2.6 19.1 ± 1.4 19.7 ± 1.5 <0.0001 0.12 0.19 Totl ody ft mss, g (n = 8) 77.5 ± 5.8 110.6 ± 10.4 210.8 ± 27.6 230.4 ± 23.1 <0.0001 0.23 0.85 Totl ody len mss, g (n = 8) 296.9 ± 10.6 305.2 ± 3.0 296.8 ± 15.4 286.2 ± 16.0 0.67 0.42 0.31 Adominl circumference, cm (n = 8) 18.9 ± 0.2 d 20.2 ± 0.3 c 22.1 ± 0.4 24.7 ± 0.5 <0.0001 <0.0001 0.0403 Tissue wet weights, mg/mm tiil length (n = 8) Liver (n = 7-8) 248.8 ± 12.0 258.0 ± 16.4 332.5 ± 1.0 363.2 ± 8.6 <0.0001 0.18 0.41 Kidneys (n = 7-8) 47.92 ± 1.3 54.62 ± 1.7 57.31 ± 3.1 58.62 ± 1.5 0.0299 0.0856 < 0.0001 Spleen (n = 7-8) 17.27 ± 1.1 19.53 ± 1.2 19.1 ± 0.3 16.11 ± 1.6 0.0042 0.9573 0.1704 Plsm iochemistry nlysis Ure (mmol/l) (n = 7-8) 2.74 ± 0.62 2.94 ± 0.31 2.90 ± 0.33 2.11 ± 0.12 0.1463 0.6032 0.5296 uric cid (μmol/l) (n = 7-8) 32.6 ± 5.36 45.4 ± 1.37 26.6 ± 2.2 c 16.4 ± 2.36 c 0.0014 0.0793 < 0.0001 Glucose, mmol/l (n = 5-8) 11.2 ± 0.61 12.6 ± 1.19 12.4 ± 0.45 9.7 ± 0.44 0.0118 0.4315 0.6681 Ech vlue is men ± S.E.M. Numer of repetitive experiments indicted within prenthesis. Mens within row with unlike superscripts letters,, c, d differ significntly (p < 0.05). C, corn strch diet; CRP, corn strch + red pity juice; H, high ft diet; HRP, high ft diet + red pity juice. intke (512.7 ± 8.3 nd 655.2 ± 28.7 kj/dy for oth CRP nd HRP) (Tle 1). Body weight Figure 1 displys ody weights of oese rts supplemented with red pity juice. Initilly, there ws no difference in ody weight for ll the tretment groups. After 8 weeks initition of the diet, the incresed in ody weights in high-crohydrte, high-ft diet-fed rts were significntly (p < 0.05) greter thn corn strch diet fed rts. At the end of 8 weeks, supplementtion of red pity juice filed to prevent further weight gin compred with C nd H diet-fed rts (Figure 1). Consequently, oth supplemented rts showed the trend of incresing ody weight lthough the difference ws not significnt (p < 0.05) (Figure 1). Plsm iochemistry nlysis The wet weight of the liver nd kidneys were significntly greter (p < 0.05) in the high crohydrte, highft diet-fed rts (H) compred with corn strch fed-rts 600 C H 550 CRP HRP 500 Body Weight, g 450 400 c 350 Red pity tretment 300 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Week Figure 1 Body weight of oese rts supplemented with red pity juice for 8 weeks. Dt re presented s men ± SEM, n = 5 8. Vlues mrked with different letters re significntly different t the level of p < 0.05. C, corn strch diet; CRP, corn strch + red pity juice; H, high ft diet; HRP, high ft diet + red pity juice.
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 5 of 10 (C) (Tle 1). However, red pity did not produce ny significnt chnges to liver nd kidneys wet weight. Contrrily, there were no difference in wet weight of the hert nd spleen for ll the groups (C, CRP, H nd HRP). No chnges in plsm ure concentrtion in red pity supplemented rts compred with C-fed nd H-fed rts (Tle 1). Furthermore, red pity supplementtion showed the trend of uric cid nd glucose normliztion for HRP s compred to H-fed rts (Tle 1). Conversely, plsm uric cid concentrtion ws incresed significntly in CRP rts (p < 0.05) compred with C-fed rts (Tle 1). Glucose tolernce nd heptic function Red pity supplemented groups showed impired glucose tolernce even fter 8 weeks supplementtion (Figure 2). Overnight fsting lood glucose differs significntly etween H fed rts nd C fed rts. Orl dministrtion of 2 g/kg ody weight of glucose resulted in incresed lood glucose concentrtion t 30 minutes nd 60 minutes for C, CRP, H nd HRP respectively. Then, lood glucose ws slowly clered from the lood in H fed rts s oserved t 90 minutes. At the end of 120 min, HRP rts filed to reduce lood glucose concentrtion to the sl vlue. The similr results were oserved for CRP rts s compred to C-fed rts (Figure 2). Insulin concentrtion in ll the rts were unle to e detected using cos c system (dt ws not shown). High-crohydrte, high-ft feeding resulted in elevted levels of plsm lnine trnsminse (ALT), sprtte trnsminse (AST) nd lkline phosphtse (ALP) s the mrkers of liver function (Figure 3). HRP diet feeding decresed the ALP nd ALT ctivities ut significntly incresed (p < 0.05) AST ctivity (Figure 3). Plsm ALP ctivity ws not ffected in CRP group while plsm ALT ws significntly decresed in CRP compred with C-fed rts (Figure 3). However, plsm AST ctivity ws decresed in CRP group (p <0.05) (Figure 3). Crdiovsculr function The mesurement of ex-vivo crdic function of the oese rts ws ccomplished y conducting Lngendorff isolted hert. Figure 4 demonstrtes tht high crohydrte, high ft diet-fed rts showed mrked incresed in distolic stiffness in comprison with corn strch diet-fed rts (Figure 4). Results reveled tht distolic stiffness of the hert ws reduced fter supplementtion of red pity juice in corn strch fed rts (Figure 4). However, the reduction ws not significnt in HRP rts in comprison with H rts (Figure 4). Liver nd crdiovsculr structure Histologicl evlution of liver tissues showed negligile ft vcuoles ccumultion without inflmmtory cells in 9 8 7 [Blood glucose], mmol/l 6 5 4 3 2 C CRP H HRP 1 0 0 30 60 90 120 Time, min Time P-vlue Diet Tretment Interction 120 Min 0.0224 0.15 0.19 Figure 2 Orl Glucose Tolernce Test (OGTT) t 16 weeks from C, CRP, H nd HRP rts. Dt re presented s men ± SEM, n = 5 8. End-point mens without common lphet in ech dt set significntly differ t p < 0.05. C, corn strch diet; CRP, corn strch + red pity juice; H, high ft diet; HRP, high ft diet + red pity juice.
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 6 of 10 400 350 300 250 Activity, U/L 200 c C CRP H HRP 150 c 100 50 c 0 ALP ALT AST Liver enzymes Figure 3 Plsm concentrtions of liver enzymes of oese rts supplemented with red pity juice for 8 weeks. Dt re presented s men ± SEM, n = 5 8. Vlues mrked with different letters re significntly different t the level of p < 0.05. ALP, lkline phosphtse; ALT, lnine trnsminse; AST, sprtte trnsminse. C, corn strch diet; CRP, corn strch + red pity juice; H, high ft diet; HRP, high ft diet + red pity juice. C group (Figure 5A). After eight weeks supplementtion, CRP (Figure 5B) displyed less ft vcuoles ccumultion ut demonstrted mrked heptocyte llooning with cornered nucleus. High crohydrte, high ft feeding for 16 weeks resulted in ugmented ccumultion of ft vcuoles in the heptocytes with sinusoids dilttion nd incresed inflmmtory cell infiltrtion in H group (Figure 5C). Red pity supplementtion reduces the ft vcuoles nd infiltrtion of inflmmtory cells (Figure 5D). Histologicl evlution of left ventricle tissues fter fed with high crohydrte, high ft diet for 8 weeks resulted in greter inflmmtory cells infiltrtion (Figure 5G) compred to CS rts (Figure 5E). Similrly, H rts showed higher collgen deposition nd hypertrophied crdiomyocytes (Figure 5K) thn CS rts (Figure 5I). 40 Distolic siffness, k 30 20 10 0 C CRP H HRP Figure 4 Distolic stiffness of oese rts supplemented with red pity juice for 8 weeks. Dt re presented s men ± SEM, n = 5 8. Vlues mrked with different letters re significntly different t the level of p < 0.05. C, corn strch diet; CRP, corn strch + red pity juice; H, high ft diet; HRP, high ft diet + red pity juice.
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 7 of 10 Figure 5 Histopthology of liver nd hert of oese rts supplemented with red pity juice for 8 weeks. (A-D) Hemtoxylin nd eosin stining of liver section showing heptocytes with enlrged ft vcuoles (mrked s fv ) ( 20), sinusoids dilttion (mrked s sd ) nd inflmmtory cells infiltrtion round the sinusoids (mrked s in ) ( 20). (E-H) Hemtoxylin nd eosin stining of left ventricle showing inflmmtory cell infiltrtion ( 20) (mrked s in ) s drk spots outside the myocytes. (I-L) Picrosirius red stining of left ventricle showing interstitil collgen deposition ( 20) (mrked s cd ) nd hypertrophied crdiomyocytes (mrked s hy ). A,E,I, corn strch diet; B,F,J, corn strch + red pity juice; C,G,K, high ft diet; D,H,L, high ft diet + red pity juice. After supplementtion with red pity for 8 weeks, HRP rts showed slightly greter infiltrtion y inflmmtory cells into the left ventricle (Figure 5H). However, HRP rts showed reduced collgen deposition (Figure 5L). On the other hnd, no chnges in inflmmtory cell infiltrtion were oserved in CRP rts (Figure 5F) while collgen deposition ws greter in CRP rts (Figure 5J) compred to CS rts (Figure 5I). Discussion In this study, red pity juice ws used s n intervention in order to investigte the ility of the fruit juice to reverse the normlities in ody composition, metolic, liver, kidneys nd hert function in high crohydrte, high-ft diet induced oese rts s model of metolic syndrome. Previous reserch confirmed tht rts fed with high crohydrte nd high ft diet for 8 weeks produced dominl oesity, impired glucose tolernce, hypertension, dyslipidemi, inflmmtion, endothelil dysfunction, crdic firosis together with incresed stiffness of the hert [17,18]. All these chnges closely mimic the humn metolic syndrome. Results reveled tht red pity supplementtion incresed food intke only in corn strch diet-fed control rts while no chnges were oserved in red pity juice supplementtion for high crohydrte, high-ft dietinduced oese rts nd the corn strch diet fed control rts. In other words, the intervention does not ffect stiety of oese rts s the quntity of food tken ws shown to e regulted y the fctors involved in stiety perception [21]. There ws report suggesting tht the food intke ws influenced y the sensitivity of the rts to the pltility of the food in the diet [22]. Therefore, it ws proposed tht the len control rts in the present study were more sensitive to the pltility of red pity juice s compred to high crohydrte, high-ft diet-induced oese rts. Food rich in sugr nd ft is n exmple of pltle foods tht inhiit the stiety signls nd up regulte hunger senstion [23]. Despite of tht, energy intke ws incresed significntly for red pity supplemented rts suggested tht red pity contins higher mount of energy-supplying mcronutrients. Red pity dded up to the totl energy content of the diet, leding to progressive increment in ody weight, nd totl ody ft throughout the intervention period. A growing ody evidences showed tht eting fruits cn reduce energy density nd reduce energy density is ssocited with lower energy intke nd decrese ody weight [24]. Furthermore, Flood- Ogy nd Rolls [25] indicted tht consumption of fruits efore mel cn increse stiety nd reduce energy
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 8 of 10 intke ut the effects re different for different forms of fruits. Whole fruit is relted to more chewing while fruit juices do not require chewing t ll. The mount of chewing is then linked to cephlic-phse responses tht control the food intke [26,27]. This my explin the reduce risk of type 2 dietes mellitus following consumption of specific whole fruits prticulrly grpes, pples, nns, nd lueerries in prospective longitudinl cohort study mong women nd men from Nurses Helth study nd Helth Professionls Helth Study respectively [28]. It is fscinting to ddress the suppression effects of red pity on ody ft for HRP rts. As the diet ws continue for 8 weeks, supplementtion of red pity did not result in further weight gin for oese rts. A current review on regultion of ody ft mss in oesity indicted tht ody resisted ft loss once oesity is estlished [21] proly through dptive mechnism of incresing metolic rte. Contrdictory, totl ody ft mss in corn strch fed rts were significntly elevted when supplemented with red pity. It could e the norml physiologicl responses upon incresing energy intke in red pity supplemented rts. In conjunction with tht dominl circumference were incresed in CRP nd HRP rts. A possile explntion for this might e tht the increse in ft mss in red pity fed rts ws deposited in dominl region, hence it cn e detrimentl to their helth. It ws expected to see no chnges in len mss for red pity supplemented rts s len mss is metoliclly ctive tissues required for mintining sic cell function, thus ws not ffected y tretment. Liver nd kidneys from H fed rts hd higher wet weightsscompredtocfedrtssoservedinprevious study [17] while no chnges in wet weight of spleen nd herts. Likewise, recent study presented liver normlities comprises of higher liver weight, increses in lnine trnsminse (ALT) nd sprtte trnsminse (AST) nd the occurrence of inflmmtory infiltrtes in rts while renl normlities were evidenced from the incresed in weight, ft ccumultion, nd glomerulr sclerosis with consumption of high ft diet [29]. A proly explntion for this defects is through the deposition of lipid in the liver upon fructose exposure which in turn conquer most of the heptocytes volume nd upset the liver structure nd function [30]. Besides, some glucose trnsporter like GLUT2 my e involved in the trnsport of fructose to the kidney [31]. Besides, red pity-fed rts showed trend of reduction in stiffness of the hert. Previous studies reported tht distolic stiffness, crdic firosis, nd elevted superoxide production were directly ssocited with ech other [18]. Therefore, it could e tht the ccumultion of interstitil crdic collgen ws reduced [32] following red pity supplementtion, hence showed the trend of meliorting crdic stiffness. To hve etter insight of liver function, the enzymes ctivities were mesured. The collective mesurement of lkline phosphtse (ALP), lnine trnsminse (ALT) nd sprtte trnsminse (AST) reflects the severity of liver dmge. ALP is induction enzyme tht is minly found in the cell memrne of the liver nd the elevtion of this enzyme indictes primry heptic disese like cholesttic liver disese. Although the increse in ALP cn e result from extr-heptic sources like one mrrow nd skeletl muscle, the effects ws miniml. Whilst, oth ALT nd AST re lekge enzymes, nd their elevtion indicte significnt heptocellulr dmge. The possile cuses my vry including toxicity, inflmmtion, hypoxi nd tissue trum [33]. However, AST is lso found in hert, kidneys, rin, nd skeletl muscle nd hd een used s nonspecific mrker for myocrdic infrction [34]. Results reveled tht red pity juice tretment significntly reduced ALP nd ALT ut cused significnt increment in AST. Therefore, the present results suggest tht red pity juice meliorted liver dmge due to high crohydrte, high ft feeding. The reduction of liver enzymes fter the intervention proly indicted the decrese in deposition of ft nd the degree of necrosis in liver cells [35]. In contrst, the elevted level of AST could e due to myocrdil dmge s the hert muscle is rich in minotrnsferse enzymes especilly AST. The decrese in ALT hs een shown to hve protective effects from deth dietes nd Ischemic hert disese in dults from Ntionl Helth nd Nutrition Exmintion Survey (NHANES) III [36]. On the other hnd, the incresed in ALT hs een ssocited with systemic nd heptic insulin resistnce, long with increse in the secretion of insulin nd reduces insulin clernce from the liver [37]. Hence, the present study showed tht red pity juice is n importnt meditor for ttenution of liver injury. Histopthologicl evidence of liver nd hert section fter supplementtion with 5% red pity juice reveled mild improvement in heptic vcuoltion nd no inflmmtion ws seen during microscopic exmintion. Also, the collgen deposition ws reduced in left ventricle of the hert. This results ws in line with the reduce ALP nd ALT enzymes indictive of improvement in liver injury s well s consistent with slightly reduction in distolic stiffness. Previous report indicted tht intervention with 5% purple crrot juice ws lso le to reverse the structurl dmge in the liver cused y high ft feeding y showing miniml signs of microvesiculr stetosis [18]. Conclusion In summry, the present dt provide scientific evidences tht red pity juice my provide protection ginst liver dmge nd my reduce the stiffness of the hert. Red pity juice contined multiple ioctive compounds which might ct synergisticlly to produce the
Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 9 of 10 protective effects. However, the effects of the specific constituents re not investigted in the present study. Therefore, it is suggested to conduct future study in order to elucidte the possile ctive compounds responsile for the effects. Besides, further study with higher doses of red pity in diet-induced oesity model cn e conducted in future to exmine the ility of this fruit for the tretment of oesity relted diseses. It is lso crucil to tke into considertion the timing of fruit juice consumption. This is ecuse eting fruit efore mels my reduce energy intke y mens of incresing fier intke. The therpeutic effects of specific fruits should e investigted rther giving generl recommendtion to the generl popultion. Competing interests The uthors declre tht they hve no competing interests. Authors contriution NSR: preprtion of the drft, contriution to conception, design, cquisition of dt, nlysis nd interprettion of dt. LB: contriution to conception, design of the study nd interprettion of dt PI: contriution to the interprettion of dt. AR: dt nlysis nd finl pprovl of the version for pulishing. All uthors red nd pproved the mnuscript. Acknowledgements We thnk Universiti Putr Mlysi for funding the reserch ttchment t University of Southern Queenslnd, Austrli. We thnk Associte Professor Leigh C. Wrd from School of Chemistry nd Moleculr Biosciences, The University of Queenslnd, Brisne for the cquisition of dul-energy X-ry sorptiometry nd nutritionl dvice, Dr. Hemnt Poudyl nd Dr. Sunil Pnchl, Dr. Wong Weng Yew nd Mr. Mhrshi Bhswnt, University of Southern Queenslnd, Toowoom for their ssistnce with the study. Author detils 1 Deprtment of Nutrition nd Dietetics, Universiti Putr Mlysi, Serdng 43400 UPM, Mlysi. 2 School of Helth, Nursing nd Midwifery, University of Southern Queenslnd, Toowoom QLD 4350, Austrli. 3 Deprtment of Biomedicl Sciences, Universiti Putr Mlysi, Serdng 43400 UPM, Mlysi. Received: 27 Jnury 2014 Accepted: 6 June 2014 Pulished: 12 June 2014 References 1. World Helth Orgniztion: Helth Fct Sheet; 2010. http://www.who.int/ medicentre/fctsheets/fs311/en/ 2010, July, 26. 2. Bullà MN, Css-Agustench P, AmigÃ-Correig P, Arncet J, Sls-Slvdà J: Inflmmtion, oesity nd comoridities: the role of diet. Pulic Helth Nutr 2007, 10(10A):1164 1172. 3. Knk G, Aklin A, Dokumcioglu A, Ozcelik E, Bl C: Crdiovsculr risk ssessment in ptients with type 2 dietes mellitus nd metolic syndrome: role of iomrkers. Dietes Met Syndr Clin Res Rev 2011, 5(1):7 11. 4. Rolls BJ: Plenry Lecture 1 Dietry strtegies for the prevention nd tretment of oesity. Preceedings Nutrit Soc 2010, 69:70 79. 5. 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Rmli et l. BMC Complementry nd Alterntive Medicine 2014, 14:189 Pge 10 of 10 35. Wng L, Meng X, Zhng F: Rserry ketone protects rts fed high-ft diets ginst nonlcoholic stetoheptitis. J Med Food 2012, 15(5):495 503. 36. Schooling CM, Kelvin EA, Jones HE: Annls of Epidemiology Alnine trnsminse hs opposite ssocitions with deth from dietes nd ischemic hert disese in NHANES III. Ann Epidemiol 2012, 22(11):789 798. 37. Bonnet F, Ducluzeu P, Gstldelli A, Lville M, Anderwld CH, Konrd T, Mri A, Blku B: Liver enzymes re ssocited with heptic insulin resistnce, insulin secretion, nd glucgon concentrtion in helthy men nd women. Dietes 2011, 60:1660 1667. doi:10.1186/1472-6882-14-189 Cite this rticle s: Rmli et l.: Effects of red pity juice supplementtion on crdiovsculr nd heptic chnges in high-crohydrte, high-ft diet-induced metolic syndrome rts. BMC Complementry nd Alterntive Medicine 2014 14:189. Sumit your next mnuscript to BioMed Centrl nd tke full dvntge of: Convenient online sumission Thorough peer review No spce constrints or color figure chrges Immedite puliction on cceptnce Inclusion in PuMed, CAS, Scopus nd Google Scholr Reserch which is freely ville for redistriution Sumit your mnuscript t www.iomedcentrl.com/sumit