Oxidative Stress During Rehabilitation from Protein Malnutrition Associated with Aerobic Exercise in Rats

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1 45 Vol.50, n. 1 : pp.45-55, Jnury 2007 ISSN Printed in Brzil BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY A N I N T E R N A T I O N A L J O U R N A L Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted with Aeroic Exercise in Rts Frncisco José Andriotti Prd 1*, Denise Vz Mcedo 2 nd Mri Alice Rostom de Mello 3 1 PPGEF; Ctholic University of Brsíli - UCB; Brsíli - DF - Brsil. 2 Lortório de Bioquímic do Exercício - Lex; Universidde de Cmpins - UNICAMP; São Pulo - SP - Brsil. 3 Deprtmento de Educção Físic, - Universidde Estdul Pulist - UNESP; Rio Clro - SP - Brsil ABSTRACT This study ws designed to evlute iomrkers of oxidtive stress in rts with or without eroic exercise during recovery from protein mlnutrition. From the 30 th to the 90 th dy of life, mle Wistr rts were fed low protein diet (LP + 6%) followed y norml protein diet (NP = 17%) until the 120 th dy nd seprted in two groups: sedentry (S) nd exercise trined (E = swimming 1h/dy, 5 dys/week, with from the 90 th to the 120 th dy). Rts fed norml protein diet were used s controls. Results showed tht physicl exercise hd eneficil effects on ody weight gin during nutrition rehilittion. Erythrocytes ctlse nd glutthione reductse (iomrkers of the ntioxidnt system) were significntly reduced in ll groups in comprison to the sedentry control group. The plsm concentrtion of TBARs (iomrkers of the oxidtive dmge) ws lso lower in the recovered rts, suggesting tht the improvement in ody growth fter nutritionl rehilittion with physicl exercise could e relted to decrese in the oxidtive stress level. Key words: Protein mlnutrition, nutritionl recovery, exercise, oxidtive stress INTRODUTION Exercise trining hs mny physiologicl effects consistent with improved helth. Severl studies hve shown tht exercise is ssocited with incresed oxidtive cpcity of skeletl muscle (Pels et l., 1989) nd muscle growth (Wtt et l., 1982), improvement in crdic function (Perez et l., 2003) enhnced one minerliztion (Bker nd Griminger, 1983) nd incresed life spn (Goodrick, 1980). Physicl exercise my lso e eneficil prt of rehilittion from mlnutrition. Torun nd Viteri (1984) reported tht physiclly ctive rts grew more in length nd weight thn the inctive counterprts lso. These uthors reported tht months old children under tretment for severe protein mlnutrition were ssigned to either n ctive group (stimulted, ut not forced to prticipte in gmes nd ctivities tht involved wlking uphill, climing rmp running, nd climing strs) or control group (with d liitum pttern of physicl ctivity nd rest oserved in child cre nd nutrition rehilittion centers). The ctive group grew more in length nd len ody mss thn the control group (Torun nd Viteri, 1984). Incresed energy demnd during physicl exercise induced increse in oxygen supply to ctive tissues (Sen, 2001). These resulted in n incresed production of rective oxygen species (ROS), minly due to elevted rtes of mitochondril respirtory chin (Sjödin et l., 1990; Sen, 2001). * Author for correspondence Brzilin Archives of Biology nd Technology

2 46 Prd, F. J. A. et l. When ROS production overcomes the cpcity of the ody ntioxidnt system, n oxidtive stress occurs nd mny cellulr constituents such s lipids, proteins nd DNA my suffer oxidtion due to ROS ttck, compromising cell function. It hs lso een suggested tht kwshiorkor, severe form of protein mlnutrition, my itself e product of the rdicl initited tissue dmge in the mlnourished child (Golden nd Rmdth, 1987; Mnry et l., 2000). The present study ws designed to evlute iomrkers of oxidtive stress, nutritionl sttus nd muscle metolism in rts sumitted or not to exercise during month recovery from protein mlnutrition. METHODS AND MATERIALS Animls nd diets Young (30 dys) mle Wistr rts were fed isocloric (3948 kcl/kg) norml (NP= 17%) or low (LP=6%) protein diets d liitum. Tle I shows the composition of oth diets. The norml protein diet followed the Americn Institute of Nutrition AIN-93G recommendtions for rodents (Reeves et l., 1993). Detiled minerl nd vitmin mix composition were descried elsewhere (Reeves et l., 1993). Tle 1 - Diet composition (g/kg). Components Norml protein (AIN-93) Low protein Csein (84% protein) Cornstrch Dextrinized cornstrch Sucrose Soyen oil L-cystine 3 1 choline chloridrte minerl mix (AIN-93) vitmin mix (AIN-93) Fier (mycrocellulose) Reeves et l., (1993). Experimentl groups Control Norml protein sedentry [NP-S]: rts fed the norml protein diet for 90 dys nd kept sedentry (n = 15); Norml protein exercised [NP-E]: rts fed the norml protein diet for 90 dys nd sumitted to swimming for one hour/dy, 5 dys week, supporting lod equivlent to 5% of ody weight, in tnk filled with wter t 30 ± 2 C, during the lst 30 dys (n = 10); Recovered Low protein/norml protein sedentry [LP/NP- S]: rts fed the low protein diet for 60 dys nd the norml protein diet for 30 dys, kept sedentry (n = 15); Low protein/norml protein exercised [LP/NP- E]: rts fed the low protein diet for 60 dys nd the norml protein diet for 30 dys ssocited with swimming exercise (n = 10). Tissue Smples At the 90 th dy of experiment, rts from ll groups were nesthetized without previous fsting with n intrperitonel injection of chlorl hydrte 10% (wt/vol), 0.3 ml for ech 100 g of ody weight for tissue nd lood extrction. They were used in experiments s soon s nesthesi ws ssured y loss of pedl nd cornel reflexes. Heprinized lood smples were otined from portl vein, for preprtion of hemolyste nd for plsm seprtion. After lood collection, the nimls were scrificed for other iologicl mteril otinment. At the 60 th dy of tretment rts of the NP-S nd of the LP/NP-S groups were scrificed for iochemicl nlysis relted to nutritionl sttus evlution. Soleus Muscle Glucose Metolism For mesurements of muscle glucose uptke nd oxidtion, glycogen synthesis nd lctte production, longitudinl soleus muscle strips (25- Brzilin Archives of Biology nd Technology

3 Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted mg) were plced in glss vils contining d [U14 C] glucose, (0.25 µci/µmol glucose), [3H]- 2-deoxyglucose (0.1 µci/µmol glucose) nd non rdioctive glucose to finl concentrtion of 5.5 mmol/l plus insulin (100miU/mL). Glucose uptke nd oxidtion s well s glycogen synthesis nd lctte production were determined s previously descried (Prd et l., 2003; Nunes et l., 2005). Enzyme Assys The hemolyste ws diluted 1:20 in 0.1M phosphte uffer, ph 7.0 efore ssying. All enzyme ssys were performed in triplicte, ccording to the following methods. Ctlse (CAT) ctivity ws mesured in 50 mm phosphte uffer (ph 7.0) y monitoring the decrese in sornce t 240 nm for 30 sec fter the ddition of 10 mm hydrogen peroxide (Aei, 1984). One unit of ctlse ctivity is the mount of enzyme present tht decomposes 1 µm H 2 O 2 /min t 25 o C (Aei, 1984). The erythrocyte CAT ctivity ws expressed s IU/g H.min. Glutthione Reductse (GR) ctivity ws mesured in 0.2 M phosphte uffer (ph 7.0) contining 2 mm EDTA, 0.1 mm NADPH nd 0.75 mm DTNB. GSSG (1mM) ws dded into the cuvette to strt the rection nd the sornce ws followed t 412 nm for 3 min (Smith, et l., 1988). The erythrocyte GR ctivity ws expressed s IU/g H (Beutler, 1975). Plsm Rective Cronyl Derivtives The plsm rective cronyl derivtives (RCD) ws mesured y cronyl regent 2.4- dinitropheylhydrzine (DNPH) using spectrophotometric method (Eston Finney et l., 1996). Plsm (200 µl) ws mixed with 1 ml H 2 O nd 2 ml of 20% triclorocetic cid (TCA) nd centrifuged t 1,000 g for 10 min. The pellet ws suspended in 1 ml of 10 mm DNPH nd incuted for 60 min t 37 o C. For lnk 1 ml of 1M HCl ws used insted DNPH. Susequently, 1mL of 20% TCA ws dded nd the smple ws centrifuged t 1,000 g for 10 min. The pellet ws wshed with 1:1 ethnol/ethyl cette solution nd centrifuged t 1,000 g for 10 min. The pellet ws mixed with 1 ml of 6 M gunidine (diluted in 20 mm H 2 PO 4, ph 2.3). Finlly the smple ws incuted for 40 min t 37 o C. The sornce ws mesured t 380 nm. The results were expressed s µmols RCD/mL of plsm The nlyses were done in triplicte nd the men ws used to sttisticl nlysis. Plsm Thiorituric Acid Rective Sustnces Plsm Thiorituric Acid Rective Sustnces (TBARs) were mesured y fluorimetry, ccording YAGI (Ygi, 1994). Plsm (20 µl) ws mixed with 4.0 ml of N/12 H 2 SO 4. Phosphotungstic cid 10% (0.5 ml) ws dded to this mixture, which ws centrifuged t 3,000 rpm for 10 min. The pellet ws mixed with 2.0 ml of N/12 H 2 SO 4 nd 0.3 ml of 10% phosphotungstic cid nd centrifuged gin t 3,000 rpm for 10 min. Distilled (4.0 ml) wter nd 1.0 ml of thiorituric cid (TBA) (1: % TBA queous solution nd glcil cetic cid) suspended the pellet. This mixture ws heted t 95 C for 60 min. After cooling, 5.0 ml of n- utnol ws dded, nd the mixture, shken vigorously. After centrifugtion t 3,000 rpm for 15 min, the n-utnol lyer ws mesured t 553 nm with excittion t 515 nm. Nutritionl Sttus Evlution For nutritionl sttus evlution, plsm glucose, totl protein, lumin nd free ftty cids (FFA) levels were mesured y colorimetric methods, s descried y Nogueir et l (1990). Liver lipids were extrcted with ethnol (Prd et l., 2003) nd mesured y colorimetric method (Nogueir et l., 1990). Sttisticl Anlysis The results were expressed s mens ± SD for the numer of rts (n) indicted. When working with muscle strips, n refered to the numer of strips. The dt were nlyzed y Student s unpired t- test or ANOVA, where dequte, nd P < 0.05 vlues indicted significnt difference. RESULTS Effects of the low protein diet on nutritionl sttus Tle 2 shows the results on plsm mrkers of nutritionl sttus fter 60 dys feeding the LP diet compred to NP group. The LP group showed significntly reduced ody weight gin, hypoproteinemi, hypoglycemi, hypoluminemi, high circulting FFA nd liver lipid levels. Despite Brzilin Archives of Biology nd Technology

4 48 Prd, F. J. A. et l. of the low ody weight gin oserved in the LP nimls, no difference in food intke ws seen etween LP nd NP. Tle 2 - Body weight gin (g), food intke (g/100g ody weight), plsm glucose (mg/dl), plsm protein, plsm lumin (g/dl), plsm FFA (µeq/l) nd liver lipid levels fter 60 dys of mlnutrition Groups NP LP Body weight gin ± ± 3.94 * Food intke 20.5 ± ± 4.50 Plsm glucose 122 ± ± 4.2 * Plsm totl protein 6.4 ± ± 0.10 * Plsm lumin 5.8 ± ± 0.13 * Plsm FFA ± ± 45.4 * Liver lipid 2.6 ± ± 0.44 * Results re men ± SD from 5 rts in the ech group, except for ody weight gin nd food intke where n = 15. NP = Norml protein diet, LP = Low protein diet ( * ) significntly from NP (P < 0.05, t-test). Effects of one month of re-feeding ssocited or not to exercise on nutritionl sttus Tle 3 shows the dt for oth groups (LP nd NP) fter 30 dys of recovery feeding NP diet (LP/NP-S nd NP-S). No significnt difference in proteinemi, luminemi, circulting FFA levels nd liver lipid content etween the groups. Body weight gin ws twice when compred to tht oserved in the group one month efore (LP group, Tle 2), ut significntly lower in the protein restricted (LP/NP-S) thn in norml protein (NP-S) group. Tle 3 lso showed tht there ws significnt increse in ody weight gin in LP/NP-E compred to LP/NP-S group. The results were completely different for NP groups, with significnt decrese in ody weight gin in NP-E compred to NP-S group despite the food intke ws higher in trined (NP-E nd LP/NP-E) thn in sedentry (NP-S nd LP/NP-S) groups. Plsm glucose ws elevted in the LP/NP-E nd LP/NP-S groups in reltion to the control (NP-S nd NP-E) nimls. Plsm lumin ws lower in oth exercised groups, significntly for the LP/NP-E group. On the other hnd, proteinemi, circulting FFA levels nd liver lipid content did not chnge with exercise even in LP/NP-E group when compred to the others. Effects of low protein diet nd re-feeding ssocited or not to exercise on skeletl muscle glucose metolism Skeletl muscle glucose uptke nd glycogen synthesis were significntly higher only in trined (NP-E nd LP/NP-E) rt isolted muscle thn in the corresponding sedentry (NP-S nd LP/NP-S) ones. However, the LP/NP-E group exhiited significnt lower glycogen synthesis when compred to NP-E group. No difference in glucose oxidtion ws oserved mong the four groups. The LP/NP-E rts showed higher muscle lctte production thn LP/NP-S rts (Fig. 1). Effects of low protein diet nd re-feeding ssocited or not to exercise on iomrkers of oxidtive dmge nd ntioxidnt system Fig. 2 shows the effects of re-feeding on oxidtive stress mrkers. Plsm TBARs concentrtions were higher in the NP-E nd in LP/NP-E thn in the corresponding sedentry groups, ut it ppered significntly decresed in the recovered (LP/NP-S) rts compred to the corresponding control rts (NP-S) (Fig. 2A). Plsm RCD concentrtions were significntly incresed only in the exercised control rts (NP-E) compred to the other groups (Fig. 2B). Brzilin Archives of Biology nd Technology

5 Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted 49 mmol/g.60 min ,0 1,5 1,0 0,5 0,0 0,20 0,15 0,10 0,05 0,00 NP-S Glucose Uptke 15 Glucose Oxidtion Glycogen Synthesis Lctte Production LP/NP-S NP-E c LP/NP-E Fig. 1 - Skeletl muscle glucose uptke, glucose oxidtion, glycogen synthesis nd lctte production y soleus muscle fter 60 dys of protein restriction followed y nutritionl recovery ssocited or not to exercise for 30 dys. Results re men ± S.D. of the 10 muscles strips per group. NP = norml protein; LP = Low protein diet; S = Sedentry; E = Exercise trined. ( ) Significnt differences (P < 0.05, ANOVA) in comprison to the NP-S rts. ( ) Significnt differences (P < 0.05, ANOVA) in comprison to the LP/NP-S rts. ( c ) Significnt differences (P < 0.05, ANOVA) in comprison to the NP-E rts. Tle 3 - Body weight gin (g), food intke (g/100g ody weight), plsm glucose (mg/dl), plsm protein (g/dl), plsm lumin (g/dl), plsm FFA (µeq/l) nd liver lipid (mg/100mg) levels fter 30 dys of mlnutrition nd nutritionl recovery ssocited or not exercise for 30 dys. Groups NP-S NP-E LP/NP-S LP/NP-E Body weight gin ± ,9 ± 6, c ± c ± 4.8 Food Intke 18.1 ± ± ± ± 4.1 Plsm glucose ± ± ± c ± 1.8 Plsm protein 6.9 ± ± ± ± 0.05 Plsm lumin 4.4 ± ± c ± ± 0.1 Plsm FFA ± ± ± ± 19.8 Liver lipid levels 4.0 ± ± ± ± 0.1 Results re men ± S.D. from 10 nimls per group. NP = norml protein; LP = Low protein diet; S = Sedentry; E = Exercise trined. ( ) Significnt differences in comprison to the NP-S rts (P < 0.05, ANOVA). ( ) Significnt differences in comprison to the LP/NP-S rts (P < 0.05, ANOVA). ( c ) Significnt differences in comprison to the NP-E rts (P < 0.05, ANOVA). Brzilin Archives of Biology nd Technology

6 50 Prd, F. J. A. et l. These dt indicted tht the NP-E group ws sumitted to higher stress level thn the other groups. Fig. 3 showed tht erythrocyte CAT nd GR ctivities were significntly reduced in ll experimentl groups (NP-E, LP/NP-S nd LP/NP- E) in comprison to the NP-S group. DISCUSSION The rts fed the LP diet exhiited fetures typicl of protein mlnutrition such s low ody weight gin, hypoglycemi, hypoluminemi, high plsm free ftty cid nd high lipid levels (Torun nd Chew, 1994; Mour nd Pssos, 2002). The results showed tht fter one month of recovery with norml protein diet these metolic ltertions were reversed. Dily food intke nd ody weight gin were improved when nutritionl recovery ws ssocited to physicl exercise. These results corroorted the hypothesis tht physicl exercise cused eneficil effect on ody growth during nutritionl recovery (Torun nd Viteri, 1984). The increse in musculr work induces severl iochemicl rections essentil to muscle growth. Physicl trining hs the ility to revert the ltertions in mitochondril protein induced y ctolic sttes in rt vstolterl nd gstrocnemius muscle (Midoui et l., 1996). These spects my e relted to growth hormone (GH) levels. GH hs mny iologicl effects on ody growth ffecting protein, crohydrte nd lipid metolism nd stimulting crtilge nd one growth. It hs een lredy demonstrted tht dministrtion of GH restored ody weight nd length in protein deficient young rts (Muku et l., 1997). GH levels cn e influenced y physicl ctivity, with tendency to elevtion especilly in exercises ove the lctte threshold (Felsing et l., 1992). In the present study, physicl trining improved glucose uptke y skeletl muscle oth in control (NP-E) nd recovered (LP/NP-E) rts. This is not surprising, since physicl exercise is known to improve glucose tolernce nd sensitivity to insulin (Del, 1990; Ngsw et l., 1990). Glucose trnsport into skeletl muscle is cutely regulted y oth insulin nd y contrctile ctivity (Hendriksen et l., 1996), through the trnsloction of glucose trnsporter isoform, the GLUT-4 (Ezki, 1997). The cpcity for glucose trnsport is ssocited with the muscle content of GLUT-4 protein (Hedriksen et l., 1996; Ezki, 1997). Both insulin stimulted glucose trnsport (Hedriksen et l., 1996; Bonen et l., 1986) nd GLUT-4 protein (Bonen et l., 1986; Rodnik et l., 1992; Houmd et l., 1995) increse in the exercise trined skeletl muscle. Despite the incresed glucose uptke of skeletl muscle in LP/NP-E rts nd norml uptke in LP/NP-S rts, lood glucose concentrtion ws elevted in oth recovered groups, when compred to their control counterprts. This my e indictive of insulin resistnce in tissues other thn skeletl muscle, induced y the erly protein restriction. Besides incresed glucose uptke, other skeletl muscle dpttions to eroic trining, concerning glucose metolism re present s n incresed glycogen synthesis (Houmd et l., 1995; Bgy et l., 1978) nd improved lctte relese from muscle to lood through n incresed plsm memrne trnsport system (Pilegrd et l., 1993). In the present study, oth incresed muscle glycogen synthesis nd incresed lctte relese to the culture medium in the skeletl muscle isolted from protein restrict trined rts (LP/NP-E) were oserved. Tken together, these results indicted tht protein restriction erly in life did not impir muscle glucose metolism responses to exercise trining. Protein restriction seemed to ccentute lctte relese from the trined skeletl muscle. Peroxidtion of unsturted ftty cid residues of phospholipids in cell memrnes might result in significnt loss in memrne integrity, which ws one of the most striking effects of oxidtive dmge (Tppel, 1973), leding to genertion of potentilly hrmful ldehydes nd lknes tht rected with thiorituric cid. Therefore, the nonspecific test of thiorituric cid rective sustnces (TBARs) ws often pplied to ssess the degree of oxidtive stress (Tppel, 1973). In ddition, oxidtive ltertion of minocids hs significnt effects on cellulr function s oxidtively modified proteins modify their physiologicl ctivity nd tend to e very sensitive to proteolytic degrdtion (Stdtmn, nd Oliver, 1991). Brzilin Archives of Biology nd Technology

7 Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted c TBARs (mmol/l) NP-S LP/NP-S NP-E LP/NP-E 10 8 c RCD (µmol/l) NP-S LP/NP-S NP-E LP/NP-E Figure 2 - Plsm thiorituric cid rective sustnces (TBARs) nd Rective Cronyl Derivtive (RCD) fter 60 dys of protein restriction followed y nutritionl recovery ssocited or not to exercise for 30 dys. Results re men ± S.D. of the 10 nimls per group. NP = norml protein; LP = Low protein diet; S = Sedentry; E = Exercise trined. ( ) Significnt differences (P < 0,05, ANOVA) in comprison to the NP-S rts. ( ) Significnt differences (P < 0,05, ANOVA) in comprison to the LP/NP-S rts. ( c ) Significnt differences (P < 0,05, ANOVA) in comprison to the NP-E rts. Brzilin Archives of Biology nd Technology

8 52 Prd, F. J. A. et l Gluthtione Reductse U/g H NP-S LP/NP-S NP-E LP/NP-E CATALASE UI/H.min NP-S LP/NP-S NP-E LP/NP-E Figure 3 - Blood Glutthione Reductse nd Ctlse ctivity fter 60 dys of protein restriction followed y nutritionl recovery ssocited or not to exercise for 30 dys. Results re men ± S.D. of the 10 nimls per group. NP = norml protein; LP = Low protein diet; S = Sedentry; E = Exercise trined. ( ) Significnt differences (P < 0,05, ANOVA) in comprison to the NP-S rts. ( ) Significnt differences (P < 0,05, ANOVA) in comprison to the LP/NP-S rts. Brzilin Archives of Biology nd Technology

9 Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted 53 Oxidtive modifictions of mino cids residues include trnsformtion of mino cid residues such s proline, rginine nd lysine to rective cronyl derivtives (RCD) nd links hve een estlished etween the degree of RCD ccumultion nd vriety of physiopthologicl conditions (Stdtmn, 1992). Results showed tht plsm TBARs concentrtion decresed significntly in the protein restricted recovered groups (LP/NP-S) in reltion to the corresponding control group (NP-S). This suggested low oxidtive dmge to plsm or tissue lipids in the first group, which could result from lower ROS production or higher removl cpcity in protein restricted thn in control rts. This pprent protection of the recovered nimls could e etter explined y n efficient plsm ntioxidnt defense, reflected in this study y decrese in the plsm lumin concentrtions thn y n increse in efficiency in the enzymtic ntioxidnt system, s the enzymes GR nd CAT re significntly decresed in this group (LP/NP-S) compred to control group (NP-S). Exercise trined rts (NP-E nd LP/NP-E) showed increse in plsm TBARs concentrtions when compred to their sedentry counterprts, indicting tht trining led to n increse in the oxidtive stress level, pprently higher for control trined rts (NP-E). Single outs of exercise my led to increse in the content of TBARs (Alessio nd Goldfr, 1988) nd RCD (Rdák et l., 1998). Aeroic exercise trining, in turn my reduce TBARs (Alessio nd Goldfr, 1988) nd RCD (Rdák et l., 1999) fter single outs of exercise, ut my e ineffective in chnging significntly these iomrkers of oxidtive dmge t rest (Pereir et l., 1994; Rdák et l., 1999). These discrepncies etween the present nd previous results (Pereir et l., 1994; Rdák et l., 1999) concerning the effects of exercise trining on iomrkers of oxidtive dmge mesured of rest could e ttriuted to differences in the swimming trining protocol performed y the rts. In summry, the results presented here suggested tht the improvement in ody growth nd muscle glucose metolism fter nutritionl rehilittion with physicl exercise cn e relted with decrese in the oxidtive stress level, lthough the enzymtic ntioxidnt ctivity is lso ffected fter four weeks of trining. ACKNOWLEDGEMENTS The uthors thnk Clrice Y. Siuy, Edurdo Custódio, José Roerto R. d Silv, Aline Cristin Csonto, Amnd Pul Pedroso, Lucin de Lim nd Corn Product Brzil. Reserch supported y the Brzilin Foundtions FAPESP (proc. 00/ , 00/ nd 02/ ), FUNDUNESP (proc.00154/03-dfp) nd CNPq (proc /96-8). RESUMO O presente estudo foi delinedo pr vlir iomrcdores de estresse oxidtivo em rtos sumetidos ou não o exercício físico durnte recuperção d desnutrição protéic. Pr tnto, rtos Wistr limentdos com diet hipoprotéic (LP = 6%) dos 30 dis té 90 dis, e então com diet normoprotéic (NP = 17%) té 120 dis, form seprdos em dois grupos: sedentário (S) e exercitdo (E = ntção 1h/di, 5 di/semn, dos 90 té os 120 dis de vid). Rtos limentdos com diet normoprotéic form usdos como controles. Nossos resultdos mostrrm que o exercício físico durnte recuperção nutricionl, teve efeitos enéficos no gnho de peso corporl e no metolismo glicídico musculr. Ctlse e gluttion reductse (iomrcdores do sistem de ntioxidnte) mostrrm-se significtivmente reduzids em todos os grupos qundo comprds o grupo controle sedentário. A concentrção plsmátic de TBARs (iomrcdores do tque oxidtivo) tmém foi mis ix nos rtos recuperdos, sugerindo que melhori no crescimento corporl e no metolismo de glicose do músculo pós reilitção nutricionl com exercício físico pode estr relciond com um diminuição no nível de estresse oxidtivo. REFERENCES Aei, H. (1984), Ctlse in vitro. Meth Enzymol., 105, Alessio, H.M., Goldfr, A.H. (1988), Lipid peroxidtion nd scvenger enzymes during exercise, dpttive response to trinning. J Appl Physiol., 64, Beutler, E. (1975), Red Cell Metolism In: A mnul of iochemicl methods, 2 ed., Grune nd Strtton pulishers, London. Brzilin Archives of Biology nd Technology

10 54 Prd, F. J. A. et l. Bonen, A., Clune, P.A., Tn, M.H. (1986), Chronic exercise increses insulin inding in muscles ut not liver. Am J Physiol., 251,E193-E203. Del, F. (1990), Diminished rginine stimulted insulin secretion in trined mn. J Appl Physiol., 69, Eston Finney, R.G., Finney, S., Bker, S., Bltzopoulos, V. (1996), Muscle tenderness nd pek torque chnges fter downhill running following prior out of isokinetic eccentric exercise. J Sports Sci., 14, Ezki, O. (1997), Regultory elements in insulinresponsive glucose trnsporter (GLUT-4) gene. Biochem Biophys Res Commun., 241,E1-6. Felsing, N.E., Brsel, J.A., Cooper, D.M. (1992), Effect of low nd high intensity exercise on circulting growth hormone in mn. J Clin Endocrinol Met., 75, Golden, M.H.N., Rmdth, D. (1987), Free rdicls in the pthogenesis of kwshiorkor. Proc of the Nut Soc., 46, Hendriksen, E.J., Stump, C.S., Trinth, T.T., Bety, S.D. (1996), Role of glucose trnsport in glycogen supercompenstion in reweighted rt skeletl muscle. J Appl Physiol., 80, Houmrd, J.A., Hickey, M.S., Tyndll, G.L., Gvign, K.E., Dohm, L. (1995), Seven dys of exercise increse GLUT-4 protein content in humn skeletl muscle. J Appl Physiol., 79, Mnry, M.J., Leeuvwenurgh, C., Heinecke, J.W. (2000), Incresed oxidtive stress in kwshiorkor. J Ped., 137(3), Midoui, A.E., Tncrede, G., Ndeu, A. (1996), Effect of physicl trining on mitochondril function in skeletl muscle of norml nd dietic rts. Metolism, Clin And Exper., 45, Mour, E.G. nd Pssos, M.C.F (2002). Importnce of the Nutritionl Sttus for the Interprettion of Nucler Medicine Exmintions. Brzilin Archives of Biology nd Technology., 45-Specil, Muku, G.M., Thissen, J.P., Gerrd, G., Ktelslegers, J.M., Miter, A. (1997), Post ntl ctch-up growth induced y growth hormone nd insulin-growth fctor-1 in rts with intruterine growth retrdtion cused y mternl protein mlnutrition. Peditr Res., 42, Ngsw, J., Sto, Y., Ishiko, T. (1990) Effect of trining nd detrining on in vivo insulin sensitivity. Int J Sports Medicine., 11, Nogueir, D.M., Strufldi, B., Hirt, M.H., Adll, D.S.P., Hirt, R.D.C. (1990), Métodos de ioquímic clínic., São Pulo, PANCAST. Nunes, W. M. S., Mello, M. A. R. Glucose metolism in rts sumitted to skeletl muscle. denrvtion. Brzilin Archives of Biology nd Technology., 48, p , Pels, A.L.L., White, T.P., Block, W.S. (1985), Effects of exercise trining on plsm lipid nd lipoproteins in rts. J Appl Physiol., 58, Pereir, B., Cost-Ros, L.F.B., Sfi, D.A., Medeiros, M.H.G., Curi, R., Bechr, E.J.H. (1994), Superoxide dismutse, ctlse nd glutthione peroxidse ctivities in muscle nd lymphoid orgns of sedentry nd exercise-trined rts. Physiol. Behv., 56(5), Perez, A.C., Oliveir, A.C.C., Estevez, E., Molin, A.J., Prieto, J.G., Alvrez, A.I. (2003), Mitochondril, srcoplsmtic memrne integrity nd protein degrdtion in hert nd skeletl muscle in exercised rts. Comp Bioch Physiol Prt C., 134, Pilegrd, H., Juel, C., Wirnd, F. (1993), Lctte trnsport studied in gint srcolemml vesicles from rts, effects of trining. Am J Physiol., 264,E156- E160. Prd, F.J.A., Mcedo, D.V., Mello, M.A.R. (2003), Evlution of protein deficient diet in rts through lood oxidtive stress iomrkers. Res Communi Mol Pthol Phrmco., v , 213. Rdák, Z., Kneko, T., Thr, S., Nkmoto, H., Ohno, H., Ssvári, M., Nyks, C., Goto, S. (1999), The effect of exercise trining on oxidtive dmge of lipids, proteins nd DNA in rt skeletl muscle, evidence for eneficil outcomes. Free Rd Biol Med., 27, Rdák, Z., Nkmur, A., Nkmoto, H., Asno, K., Ohno, H., Goto, S. (1998), A period of neroic exercise increses the ccumultion of rective cronyl derivtives in living of rts. Pfluger Arch., 173, Reeves, P.G., Nielsen, F.H., Fhey, G.C. (1993), AIN- 93 purified diets for lortory rodents, finl report of the Americn Institute of Nutrition Ad Hoe Writing Commitee on the Reformultion of AIN-76A. J Nutr., 123, Rodnick, K.J., Hendriksen, E.J., Jmes, D.E., Hollozzy, J.D. (1992), Exercise trining, glucose trnsporters nd glucose trnsport in rt skeletl muscle. Am. J. Physiol., 262,C9-C14. Sen, C.K. (2001), Exercise-induced oxidtive stress nd ntioxidnt nutrients. In: Nutrition in Sports - The Encyclopedi of Sports Medicine, Mughn R J, editor., v. 7, cp. 22. Sjödin, B., Wesling, H.E., Apple, S. (1990), Biochemicl mechnisms for oxygen free rdicl formtion during exercise. Sports Med., 10, Smith, I.K., Vierheller, T.L., Thorne, C.A. (1988), Assy of glutthione reductse in crude tissue homogentes using 5,5 -dithiois(2-nitroenzoic cid). Anl Biochem., 175, Stdtmn, E.R. (1992), Protein oxidtion nd ging. Science., 257, Stdtmn, E.R., Oliver, C.N. (1991), Metl-ctlyzed oxidtion of proteins. J Biol Chem., 266, Tppel, A.L. (1973), Lipid peroxidtion dmge to cell components. Fed Prod., 32, Brzilin Archives of Biology nd Technology

11 Oxidtive Stress During Rehilittion from Protein Mlnutrition Associted 55 Torun, B., Chew, F. (1994), Protein-energy mlnutrition. In, Nutrition in Helth nd Disese, Shils, M.; Olson, J.A.; Shike, M. (eds) Modern, v.2, Phildelphi, Le nd Feiger.,. p Torun, B., Viteri, F.E. (1984), Influence of exercise on liner growth. Eur J Clin Nutr., 48(1), S Wtt, P.W., Kelly, F.J., Goldspink, D.F., Goldspink, O. (1982), Exercise induced morphologicl nd iochemicl chnges in skeletl muscles of the rt. J Appl Physiol., 53, Ygi, K. (1994), Lipid peroxides nd relted rdicls in clinicl medicine. Adv Exp Med Biol., 366,1-15. Received: My 20, 2005; Revised: Decemer 30, 2005; Accepted: Octoer 10, Brzilin Archives of Biology nd Technology

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