Pkistn Journl of Nutrition 6 (4): 399-403, 2007 ISSN 1680-5194 Asin Network for Scientific Informtion, 2007 Effects of High Crohydrte Low Ft Nigerin-Like Diet on Biochemicl Indices in Rits H.A. Ooh, C.O. Omofom, F.E. Olumese nd B. Eiy Deprtment of Medicl Biochemistry, School of Bsic Medicl Science, College of Medicine, University of Benin, Benin City, Nigeri Astrct: The effects of High Crohydrte Low Ft (HCLF) diet on iochemicl indices were evluted in rits. The diet consisting of 85% crohydrte nd 5% ft s % of totl energy in wholly compounded form ws dministered to rits. Twelve rits were rndomly divided into two groups of six rits per group. The groups were Group I fed the control diet nd the second group II ws fed with the low protein high crohydrte diet. At the end of eight weeks, the nimls in ech group were scrificed nd nlysis done on lood smples of the rits. Results showed tht the totl protein, lumin nd gloulin were significntly (p<0.05) lowered for the rits fed HCLF diet. Asprtte trnsminse EC2.6.1.1 (AST) nd Alnine trnsminse EC.2.6.1.2 (ALT) levels incresed significntly (p<0.05) while Alkline phosphtse EC.3.3.3.1 (ALP) decresed significntly (p<0.05). The iliruin level for rits on the experimentl diet ws within the norml levels. Serum electrolyte concentrtions show significnt (p<0.05) increse in sodium nd chloride ions when compred with control. Potssium reduced significntly nd icronte ions hd no significnt chnge (p<0.05). Cretinine vlues showed significnt (p<0.05) decresed levels while ure levels incresed significntly (p<0.05) Cholesterol levels were significntly (p<0.05) reduced. The lipoprotein frctions showed significntly (p<0.05) elevted HDL nd tricylglycerol levels. LDL-Cholesterol were significntly (p<0.05) reduced. The HCLF Nigerin diet promotes hypertriglyceridemi, decresed serum proteins nd incresed AST, ALT nd ure levels suggestive of heptocellulr dmge. The reduced ALP levels is suggestive of rrested one growth. The impliction of consuming this diet is discussed. Key words: High-crohydrte low ft diet (HCLF), serum lipid, serum enzymes, electrolytes Introduction The typicl Nigerin diet consists of low protein nd high crohydrte nd fier levels. Met, fish, milk nd eggs re expensive nd out of the rech of most fmilies. New reserch hs shown the effect of low protein high crohydrte diet on some iochemicl indices (Jeppesen et l., 1997; Drlene et l., 1997; Prks nd Hellerstein, 2000; Nichols et l., 2004). Lipid profile of humns consuming HCLF diet shows increses in fsting tricylglycerol (Connor nd Connor, 1997; Ktn et l., 1997). HDL-cholesterol, LDL-cholesterol nd totl fier, low-ft diets s consumed normlly in Nigeri. This present study ws designed to exmine the chnges in some iochemicl indices on rits fed wholly compounded HCLF Nigerin-like diet. Mterils nd Methods Animls nd mngement: 3 months old New Zelnd white rits (Initil men weight 1.75Kg) were used in the present study. The rits were housed in individul stinless steel niml cges with wire mesh floors to prevent cholesterol re lso reduced. In humns, coprogrphy. Light ws 12hr-light nd 12hr drk cycle hypertriglyceridemi cn e induced endogenously y nd the temperture ws uniform. The nimls were high ft diet (Austin et l., 1998) or it cn e cclimtized on growers msh (Bendel feed nd flour crohydrte induced (Knittle nd Ahrens, 1964; Reven mills BFFM Limited, Ewu, Nigeri) for two weeks. Prior to et l., 1965). A high tricylglycerol (TAG) flux rte ws the study, food nd wter ws given d liitum. oserved in sujects fed HCLF diet (Aot, 1990). The The rits were divided into two groups of six rits TAG increse oserved in humns on HCLF diet my per group, ccording to ody weight similr fter the hve een from de novo lipogenesis (Prks nd dpttion period. One group ws fed the control diet Hellerstein, 2000). nd the second group ws fed the High crohydrte Most previous studies exmining crohydrte induced low ft diet for totl period of eight weeks. hypertriglyceridemi hve used mximum of 10 dys The composition of oth diets is shown in Tle 1. feeding protocols (Nestel et l., 1970), or diets with Fresh feed ws provided on dily sis while stle simple sugrs (Huff nd Nestel, 1982) nd liquid diet remnnts were discrded fter weighing. On the devoid of ft (Schonfeld, 1970). Such diet my e verge ech rit received out 150g/feed/dy. metolized differently from whole foods contining high Clen drinking wter ws provided d li. During this 399
Ooh et l.: Effects of High Crohydrte Low Ft Nigerin-Like Diet period, feed intke, wter intke nd dry fecl output were mesured dily. Weight gin ws recorded weekly. Animl mngement nd experimentl procedures were performed in strict ccordnce with the requirements of the Ntionl Reserch Council s Guide for the use of Lortory Animls (NRC, 1985). Blood smples, collection nd nlysis: The nimls were fsted for 18hr nd seline lood smples were drwn from the rit er veins using 21-guge syringes. At the end of the feeding period, the rits were nesthetized with pentoritl (60mg/Kg ody weight). Insertion ws mde into the hert region for collection with the use of needle nd syringe. The lood smples were collected into lelled ijou ottles contining heprin s nticogulnt nd centrifuged immeditely (3,000 g for 10 minutes), to otin the serum. The serum smples were stored in the iofreezer t-10 C until nlyzed. Duplicte serum smples for ech niml group were nlyzed for Totl proteins (Brown, 1976), lumin, gloulins, (Bertl, + + - - 1974) electrolytes (N,K,Hco 3,Cl ), (Kinsley nd Schffert, 1953), lipid, (Anderson et l., 1971), Alkline phosphtse (ALP), Asprtte trnsminse (AST), Tle 2: Weight gin, feed intke, wter intke, feed efficiency nd dry Alnine trnsminse (ALT) (Moss et l., 1971) ure nd fecl output of rits in the control nd experimentl groups cretinine (Crr, 1959) were mesured using Group Control diet Experimentl Diet commercil kits (Boehringer Mnnheim). Sttisticl nlysis: Dt were expressed s men±stndrd error of the men (SEM) for ech group of rits. Comprison etween the control nd experimentl set of dt ws nlyzed y the student s t-test nd P vlues<0.05 were indictive of significnce. The sttisticl nlyses were done with INSTAT sttisticl pckge. Results Tle 1 shows the Composition of control nd experimentl diet fed to the rits. The weight gin, feed nd wter intke, feed efficiency nd dry fecl output of the rits in the control nd experimentl groups re presented in Tle 2. Sttisticl nlysis showed tht there ws significnt decrese in weight gin, feed nd wter intke nd feed efficiency in the experimentl diet when compred with the control diet. The vlues oserved for totl protein, lumin nd gloulin re given in Tle 3. The vlues were oserved for totl proteins (1.28mg/dl), lumin (0.26mg/dl) nd gloulin (1.75mg/dl). The rits on the experimentl diet hd significntly (p<0.05) lowered vlues compred to the control diet. The vlues otined for AST, ALT, ALP nd iliruin re shown in Tle 4. The AST nd ALT levels significntly (p<0.05) incresed while ALP levels significntly (p<0.05) decresed. The iliruin level for rits on the experimentl diet (0.37mg/dl) is within the norml levels nd no significnt difference ws Tle 1: Composition of Experimentl Diets Diet type ------------------------------------- Dietry Component Control HC/LFP Grri 60.0g 85.4g Fish 15.4g 8.0g *Minerl nd vitmin mix (*OPIMIX PREMIX) 1.0g 1.0g Plm oil 18.0g 5.0g Methionine 0.60g 0.60g Note: Grri is cssv sed mel commonly consumed in Nigeri nd contriuted to the fier in the Nigerin-like diet *OPIMIX PREMIX Vitmin A 8,000,000IU Copper 5gm Vitmin D 1,600,000IU Iron 20gm Vitmin E 5,000IU Iodine 1.2gm Vitmin K 2,000mg Selenium 200mg ThimineB1 1500mg Colt 200mg Rioflvin B2 4,000mg Cholin chloride 200gm Pyridoxine-B6 1500mg Anti oxidnt 125gm Nicin 15,000mg Mngnese 80gm Vitmin B12 10mg Zinc 50gm Pthothenic cid 5,000mg Biotin 20mg Folic cid 5,000mg Weight gin (g/rit) 650±8.0 120±6.0 Feed intke (g/rit/dy) 53.8 50.5 Wter intke(ml/rit/dy) 20.50±2.0 16.80±2.50 Feed efficiency (g/ody weight/g feed) 12.08 2.37 Dry fecl output (g/rit/dy) 5.24±1.14 3.25±1.02 Vlues re men±sem of six rits. Mens of the sme row followed y different letters differ significntly (p<0.05) Tle 3: Men concentrtions of totl protein, lumin nd gloulin of rits on the control nd experimentl diet Prmeters Control Diet Experimentl Diet Totl Protein 1.28±0.41 0.40±0.17 Alumin 0.26±0.15 0.07±0.02 Gloulin 1.75±0.52 1.23±0.23 Vlues re men±sem of six rits, Mens of the sme row followed y different letters differ significntly (p<0.05) Tle 4: Men concentrtions of serum enzymes nd iliruin of rits on the control nd experimentl diet Prmeters Control Diet Experimentl Diet AST(IU/dl) 2.90±0.07 8.35±1.65 ALT(IU/dl) 7.65±0.47 10.33±0.62 ALP(IU/dl) 32.5±16.48 5.0±0.21 Biliruin (mg/dl) 0.28±0.09 0.37±0.05 Vlues re men±sem of six rits, Mens of the sme row followed y different letters differ significntly (p<0.05) oserved etween the control nd experimentl diet. Tle 5 shows the level of electrolytes, cretinine nd ure in rits. The rits fed the HCLF diet hd significnt increse (p<0.05) in sodium nd chloride ions content when compred with the control (p<0.05). Potssium ions decresed significntly (p<0.05) nd icronte ions hd no significnt (p<0.05) chnge. 400
Ooh et l.: Effects of High Crohydrte Low Ft Nigerin-Like Diet Tle 5: Men concentrtions of electrolytes, cretinine nd ure of rits on the control nd experimentl diet Prmeters Control Diet Experimentl Diet Potssium 0.47±0.11 0.28±0.27 Sodium 17.0±2.55 27.00±4.14 Bicronte 3.67±0.58 6.33±0.62 Chloride 12.00±0.58 15.5±2.30 Cretinine 0.46±0.06 0.2±0.07 Ure 8.33±3.69 17.68±3.38 Vlues re men±sem of six rits, Mens of the sme row followed y different letters differ significntly (p<0.05) Tle 6: Men concentrtions of plsm lipids on rits fed the control nd experimentl diet Prmeters Control Diet Experimentl Diet Totl Cholesterol 50.18±7.75 2.80±17.5 HDL-Cholesterol 6.5±0.96 9.34±5.23 LDL-Cholesterol 32.18±7.22 13.50±5.23 Tricylgl;ycerol 10.5±7.59 28.50±4.41 Vlues re men±sem of six rits, Mens of the sme row followed y different letters differ significntly (p<0.05) The Cretinine vlues showed significnt (p<0.05) decrese while ure levels incresed significntly (p<0.05) when compred with control diet. Tle 6 shows the vlues of serum lipids of rits fed the control nd experimentl diet. Serum cholesterol levels were significntly (p<0.05) reduced. The lipoprotein frctions were lso significntly ltered. HDL nd TAG levels were significntly elevted (p<0.05) when compred with the control. LDL-cholesterol concentrtions were lso significntly reduced in the experimentl groups when compred with the control diet. Discussion The rits fed the Nigerin like HC/LPF diet showed severe reduction of ody weight. A reduction in weight gin hs een reported in similr study with low protein intke (Hys et l., 2004; Brons, 1987). The totl protein, lumin nd gloulin levels in rits fed the experimentl diet were significntly lowered. This showed tht the rits were protein mlnourished. Similr low lumin nd gloulin vlues hve een oserved in erlier studies when rits were fed protein deficient diet (Myne, 2001). Low concentrtion of plsm protein nd lumin is cler indiction of low protein diet. Rits fed the Experimentl diet hd significntly incresed AST nd ALT levels. The low protein diet led to dpttive chnges in the liver enzymes. These enzymes hve een reported to e incresed in plsm during heptocellulr injury due to the incresed permeility of the cell memrne serum enzymes (Okolie nd Osgie, 1999). The increses in plsm AST nd ALT my e linked to degenertive chnges in the liver nd kidney following the dministrtion of protein deficient diet (Mcintyre nd Roslki, 1994). The decrese concentrtion of ALP in the rits fed the experimentl diet show tht the levels of ft used in this study did not dversely interfere with the clcifiction nd other metolic ctivities s medited y ALP. The iliruin levels hd no significnt chnge in comprison to the control. This indictes tht the experimentl diet did not ffect the mintennce of the norml excretory function of the liver. Cretinine nd Ure hve een widely used to mesure excretory function especilly in conditions where there is prerenl ltertion of ure metolism s in low protein diet, (Blis, 1976). Cretinine levels for rits fed the experimentl diet decresed significntly. This is n indictor of norml kidney function. The levels of electrolytes in rits fed the experimentl diet hd significntly higher sodium nd chloride ions. Low protein diet is ssocited with edem due to increse in totl ody wter. Sodium ion nd chloride ion is distriuted principlly in the extrcellulr fluid nd so ssocited with its retention (Etukudo et l., 1999). Increse in sodium retention hs een shown to increse the risks of n individul current lood pressure (Kotchen nd Kotchen, 1997). The 86% crohydrte diet utilized in this study ws supplied y Grri ( processed form of cssv). This cssv sed crohydrte source hs high glycemic index (Euehi et l., 2005). This will stimulte insulin production nd result in higher insulin sensitivity index. Mcneel nd Mersmnn (2005), reported similr results in rts fed high crohydrte diet. Insulin lso cuses the retention of sodium ions (Volpe, 1993). This will cuse the retention of fluid which, cuses high lood pressure nd congestive hert filure. A rised insulin level my trigger the sympthetic nervous system which will cuse rteril spsm or constriction of the rteries which my consequently rise lood pressure (Kufmn et l., 1991). Significntly reduced potssium ion levels in serum were oserved in this study. An inverse reltionship etween incresed intrcellulr sodium ion nd potssium ion deficiency results in hypoklemi (Krishner et l., 1987). Potssium levels re usully mintined in very nrrow rnge. Any devition from the norml levels cn hve devstting effects on the hert nd Centrl Nervous System (CNS). Moderte potssium depletion could led to muscle ftigue (Kritensen et l., 2006). The totl serum cholesterol nd LDL cholesterol were found to e significntly lowered in the rits fed the experimentl diet while, HDL nd TAG incresed significntly. Mensink nd Ktn (1987) nd Ullmnn et l. (1991) hve shown similr chnges in helthy men nd women fed strictly controlled diet rich in crohydrte. Ktii et l. (2004), lso reported similr results in men nd women with type II dietes mellitus. 401
Ooh et l.: Effects of High Crohydrte Low Ft Nigerin-Like Diet This crohydrte induced elevtion of TAG could e Euehi, O.A.T., O.O. Blol nd Z.A. Ahmed, 2005. explined s reduced clernce of TAG rich lipoprotein nd incresed production. There is lso possiility tht reduced lipoprotein lipse ctivity on low ft diet my limit the conversion of VLDL to LDL. Mclughlin et l. (2000) hve lso shown tht high crohydrte diet mrkedly increse ftty cid synthesis nd denovo lipogenesis. A high crohydrte diet will cuse ft to e deposited, while slowing down the urning of existing ft. This is linked to greter potentil to store ft (Hudgins, 2000). The typicl Nigerin diet consists of high crohydrte nd low protein. This is consumed from erly childhood into dulthood. This predisposes the individul to the development of oesity, hypertension, liver dysfunction nd dietes t middle ge. These diseses re incresing t n lrming rte in the Nigerin populce. Longevity might e positively influenced y reduction in the percentge of crohydrte nd increse in protein nd fier content of the diet. This will prevent diseses nd enhnce good helth. References Aot, W.G., 1990. Effect of high crohydrte low ft sturted ft diet on polipoprotein B nd triglyceride metolism in Pim indins. J. Clin. Invest., 86: 642-650. Anderson, J.T., F. Grnde nd A. Keys, 1971. Effect on mn serum lipid of two proteins with different mino cid composition Am. J. Clin. Nutr., pp: 24524-530. Austin, M.A., J.E. Holknson nd K.L. Edwrds, 1998. Hypertriglyceridemi s crdiovsculr risk fctor. Am. J. Crdiol., 81: 7B-12B. Bertl, J.M., R.P. Plcko nd G.G. Grhm, 1974. Serum proteins nd plsm free mino cids in severe mlnutrition Am. J. Clin. Nutr., 27: 733-742. Blis, M.E., 1976. Uric cid metolism in Mn. Adv. Clin. Chem., 18: 213-246. Brons, D.N., 1987. A short Textook of chemicl pthology Ed., London, Hodder nd stoughon, Richrd Cly. Brown, B.A., 1976. Hemtology, London, Butterworth, 3. Crr, J.J., 1959. Determintion of Cretinine with lkline 3,5-dinitroenzote. Anl. Chem., 25: 1859. Connor, W.E. nd S.L. Connor, 1997. Should low-ft, high-crohydrte diets e recommended for everyone? N. Engl. J. Med., 337: 562-563. Drlene, M. Dreon, Hrriett A. Fernstrom, Pul T. Willims nd Ronld M. Kruss, 1997. LDL Suclss Ptterns nd Lipoprotein Response to Low-Ft, High-Crohydrte Diet in Women Arteriosclerosis, Thromosis nd Vsculr Biol., 17: 707-714. 402 Nutritive nd nti-nutritive composition of Cssv (Mnihot esculent) tuer nd lef. Eu. J. Sci. Res., 9: 41-50. Etukudo, M.H., E.O. Agedn, O.O. Akinyink nd B.O. Osifo, 1999. Plsm electrolytes, totl cholesterol, liver enzymes nd selected ntioxidnt sttus in Protein Energy Mlnutrition. Af. J. Med. Sci., 28: 81-85. Hys, N.P., R.D. Strling, X. Liu, D.H. Sullivn, T.A. Trppe, J.D. Fluckey nd W.J. Evns, 2004. Effects of n Ad Liitum low ft, high crohydrte diet on ody weight, ody composition nd ft distriution in older men nd women. Arch. Intern Med., 164: 210-217. Hudgins, L.C., M.K. Hellerstein, C.E. Seidmn, R.A. Neese, J.D. Remroli nd J. Hirsch, 2000. Reltionship etween crohydrte induced hypertriglyceridemi nd ftty id synthesis in len nd oese sujects. J. Lipid Res., 41: 595-604. Huff, M.W. nd P.J. Nestel, 1982. Metolism of polipoproteins CII, CIII1, CIII2 nd VLDL-B in humn sujects consuming high crohydrte diets. Met., 31: 493-498. Jeppesen, J., P. Schf, C.Z. Jones, M.Y. Zhon, Y.D. Chen nd G.M. Reven, 1997. Effects of low ft, high crohydrte diets on risk fctors for ischemic hert disese in postmenopusl women. Am. J. Clin. Nutr., 65: 1027-1033. Ktn, M.B., S.M. Grundy nd W.C. Willett, 1997. Beyond low-ft diets. N. Engl. J. Med., 337: 563-566. Ktii, I.A., A.A. Aknde nd A.K. Slmi, 2004. Lipid normlities mong type II dietes mellitus ptients: our experience in Ilorin, Nigeri Med. Edu. Rresource Af., 14: 254-255. Kufmn, L.N., M.M. Peterson nd S.M. Smith, 1991. Hypertension nd sympthetic hyperctivity induced in rts y high ft nd glucose diets. Am. J. Physiol. Endocrinol. Met., 260: E95-E100. Kinsley, G.S. nd R.R. Schffert, 1953. Direct micro determintion of sodium, potssium nd clcium Anl. Chem., 25: 1738. Knittle, J.L. nd E.K.J. Ahrens, 1964. Crohydrte metolism in two forms of hyperglyceridemi. J. Clin. Invest., 43: 485-495. Kotchen, T.A. nd J.M. Kotchen, 1997. Dietry sodium nd lood pressure: interctions with other nutrients Am. J. Clin. Nutr., 65: 708. Krishner, G.G., P. Chusid nd R.D.L. Hoeldtre, 1987. Mild potssium depletion promoted renl sodium retention. J. L. Clin. Med., 109: 724-730. Kritensen, M., T. Hnsen nd C. Juel, 2006. Memrne proteins involved in potssium shifts during muscle ctivity nd ftigue. Am. J. Physiol. Regultory Integrtive Comp. Physiol., 290: 766-772. Myne, P.D., 2001. Clinicl chemistry in Dignosis nd tretment 6th Edition ELST Arnold, London, pp: 280-348.
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