Abstract
Physical exercise is important in preventing and adjunctive therapy certain diseases. However, over physical exercise for beginners may cause oxidative stress and muscle injury leads to generate chemoattractants whic enables to attract neutrophil and monocyte towards the injury and stimulates leucocytes activation. The use of antioxidant-rich fruits to mitigate exercise induced oxidative stress has been applied. This study was conducted to evaluate the effect of guava juice (Psidium guajava L) consumption during aerobic exercise on leucocytes, neutrophils and superoxydes dismutase (SOD) level of beginners. Sixteen students of National Land College (Sekolah Tinggi Pertanahan Nasional/STPN), Yogyakarta who met the inclusion and exclusion criteria were involved in this study. Subjects were divided into two groups i.e. a group given mineral water as control and another group given 240 mL guava juice 0.67 g/mL daily for 27 days as treatment group. Both groups then underwent aerobic exercise for 30 minutes every day for 27 days. Blood samples were taken before and after exercise on day 1, 7 and 14 for leucocyte and nutrophil count as well as SOD analysis. The leucocyte count and SOD level before and after exercise in both Guava juice and Mineral water groups were not significantly different (p>0.05). However, the percentage of neutrophil on day 1 and 7 were significantly higher than that in Mineral water group (p<0.05). Moreover, during the exercise, the percentage of neutrophils in Guava juice group significantly increased (p<0.05), whereas in Mineral water group was not significantly different (p>0.05). It can be concluded that consuming
and Mineral water groups were not significantly different (p>0.05). However, the percentage of neutrophil on day 1 and 7 were significantly higher than that in Mineral water group (p<0.05). Moreover, during the exercise, the percentage of neutrophils in Guava juice group significantly increased (p<0.05), whereas in Mineral water group was not significantly different (p>0.05). It can be concluded that consuming guava juice during exercise for the beginners does not influence leucocyte count and SOD levels. However, it can increase percentage of neutrophil. ABSTRAK Latihan fisik penting dalam mencegah dan terapi pendamping untuk penyakit tertentu. Namun demikian, latihan fisik berlebihan bagi pemula kemungkinan bisa menyebabkan stres oksidatif dan luka otot yang dapat menghasilkanchemoattractantsuntuk menarik neutrofil, dan monosit ke daerah luka dan juga mengaktivasi leukosit. Penggunaan buah yang kaya antioksidan untuk mengurangi stres oksidatif akibat latihan fisik telah banyak dilakukan. Penelitian ini dilakukan untuk mengkaji pengaruh pemberian jus jambu merah pada latihan aerobik pemula terhadap leukosit, neutrofil dan kadar superoxydes dismutase (SOD) darah. Enam belas mahasiswa dari Sekolah Tinggi Pertanahan (STPN), Yogyakarta yang memenuhi kriteria inklusi dan eksklusi dilibatkan dalam penelitian. Subjek dibagi menjadi dua kelompok yaitu kelompok kontrol yang diberi air mineral dan kelompok perlakuan yang diberi jus jambu merah 0,67 g/mL sebanyak 240 mL setiap hari selama 27 hari. Kedua kelompok selanjutnya melakukan aerobik selama 30 menit setiap dua hari sekali selama 27 hari. Sampel darah diambil sebelum dan setelah aerobik pada hari ke 1, 7 dan 14
54 J Med Sci, Volume 46, No. 2, June: 53 - 60 untuk pemeriksaan leukosit, neutrofil dan kadar SOD darah. Jumlah leukosit dan kadar SOD darah sebelum dan setelah aerobik pada kelompok yang diberi jus jambu merah dan air mineral tidak berbeda nyata (p>0.05). Namun demikian, persentase neutrofil pada hari ke 1 dan 7 aerobik lebih tinggi nyata dibandingkan dengan air mineral (p<0.05). Selin itu, selam aerobik persentase neutrofil pada kelompok yang diberi jus jambu merah naik secara nyata (p<0.05), sedangkan pada kelompok yang diberi air mineral tidak berbeda nyata (p>0.05). Dapat disimpulkan bahwa konsumsi jus buah merah selama latihan fisik bagi pemula tidak mempengaruhi leukosit dan kadar SOD secara nyata, tetapi dapat meningkatkan persentase neutrofil. Keywords: aerobic exercise - guava juice - leucocytes - neutrophils - SOD level INTRODUCTION Numerousstudieshaverevealedarelationship between regular exercise and improvements in physical and mental healths. Physical exercise is important in preventing and adjunctive therapy certaindiseases.Thebeneficialeffectsofexercise are associated with metabolic improvement and neutralizationofsomeriskfactorsconnectedwith diseases. 1,2 However,itshouldbeconsideredthat overphysicalexercisemaycausemuscledamage called delayed onset muscle soreness (DOMS) due to mechanical trauma followed by oxidative stress. 3 The DOMS is often experienced by the beginners. Over contraction and unusual use of muscleonthebeginners duringphysicalexercise need additional unit motor recruitment. Bone musclecontractionisamechanical,chemical,and electrical process consisting of six stages called crossbridgecycle.Forthebeginnersoruntrained people, power stroke, sliding filament, and disconnectingincrossbridgecyclearemechanical trauma which may cause immune dysfunction, inflammation,oxidativestress,musclesorenessand muscleinjury. 4-6 The muscle injury can generate chemo- attractantswhichenablestoattractneutrophiland monocyte towards the injury site followed by reactive oxygen species (ROS) production. 6,7 Moreover, oxidative stress due to over exercise causes the increase of body temperature and stimulates leucocytes production and activation. In physiological conditions, the ROS production willbeneutralizedbyendogenousantioxidantsuch as superoxydes dismutase (SOD), glutathione peroxides (GPx), and catalase. Oxidative stress willoccurwhentheendogenousantioxidantisnot enoughtoneutralizeROSproduction. 8,9 Recently, there is growinginterest in the use ofantioxidant-richfruitsorvegetablestomitigate exerciseinducedoxidativestress.Severalfruitsor vegetables have been studiedfor their benefits as dietary supplements and sport nutrition foods in countering oxidative stress during physical exercise. 10 Psidium guavaL. is a fruit tree commonly known as guava orjambu bijiin Indonesai,whichbelongstothefamilyMyrtaceae. Guava fruit isrich antioxidantsuch asvitamin C, lycopeneandflavonoids.Guavafruitisalsoagood source of pectin, sugar and minerals. 11-13 Guava fruit has been reported to have antidiarrheal, antibacterial
use ofantioxidant-richfruitsorvegetablestomitigate exerciseinducedoxidativestress.Severalfruitsor vegetables have been studiedfor their benefits as dietary supplements and sport nutrition foods in countering oxidative stress during physical exercise. 10 Psidium guavaL. is a fruit tree commonly known as guava orjambu bijiin Indonesai,whichbelongstothefamilyMyrtaceae. Guava fruit isrich antioxidantsuch asvitamin C, lycopeneandflavonoids.Guavafruitisalsoagood source of pectin, sugar and minerals. 11-13 Guava fruit has been reported to have antidiarrheal, antibacterial and antioxidant activities. 13,14 Commercially, the fruit is consumed raw or used inmakingjams,jellies,pastesandjuice. 11 In this study, we reported the effect of guava juice consumption during aerobic exercise on leucocytes,neutrophilsandsuperoxydesdismutase (SOD) level of beginners. MATERIALS AND METHODS Subjects This was a quasi experimental study with nonrandomizedpre-posttestcontrolgroupdesign involving students from National Land College (Sekolah Tinggi PertanahanNasional/STPN),
55 Ulvie et al., The leucocytes, neutrophils and superoxydes dismutase (SOD) level after consuming guava juice (Psidium guajava L) during aerobic exercise for beginners Yogyakarta who met inclusion and exclusion criteria.Theinclusioncriteriawerestudentsaged 17-25 years who stay in dormitory, normal nutritional status, normal healthy status based on clinicalandhemoglobinexaminations,nonathletes, nosufferedfromchronicdiseasessuchasasthma, hipertension,cardiovasculardiseases,renaldisease, andgartritis,nosmoking,andneverfollowedthe aerobic exercise program based on frequency, intensity,time,andtype(FITT)guidelinesminimally in the last 6 years. The exclusive criteria include consuming antioxidant and or vitamin and not willingtoberesearchsubject.Thestudyhasbeen approved by the Medical and Health Research EthicsCommittee,FacultyofMedicine,Universitas Gadjah Mada,Yogyakarta. Procedure of study Onthedaythat hasbeenagreed,the students weregatheredtobeselected.Anexplanationabout objectives,benefitsandriskofthestudywasgiven. Students were then given opportunities to ask eveythingrelatedtothestudy.Sixteenstudentswho fulfilledthe inclusionandexclusioncriteria were recruited in this study and given an informed consent to be signed. All subjects underwent physical and clinical as well as laboratory examinations.Characteristicsofsubjectsincluding bodyweight(BW),bodyheight(BH),sistolicand diastolicbloodpressure(SBPandDBP),hearrate (HR), respiratory rate, body temperature, haemoglobin level, leucocyte and neutrophils counts wererecorded.Subjectsweredividedinto twogroups with 8 students in eachgroup. Group IascontrolwasgivenmineralwaterandGroupII astreatmentgroupgivenwasgiven240mLguava juice0.67g/mLdailyfor27days.Theguavajuice was given dailay at 05.00 AM. Subjects then underwent aerobic exercise every two days for 27 days consisting 30 minute for exercise, five minutes for warming up and five minutes for recovery. Body weight, blood pressure, HR, temperature of subjects before and after aerobic exercise were monitored and recorded. Food intake of subjects were calculated based on 24- hour food recall for 27 days using nutrisurvey program.Bloodsamplesofsubjectsafterexercise on day 1, 13 and 27 were collected for leucocyte andneutrophilcountaswellasSODexamination. Leucocyte andneutrophil counts were measured using HematologyAnalyzer Sysmex XT-2000i, whereas blood SOD level was measured using caymankit. Statistical analysis Data were presented as mean ± standard deviation (SD). The different of leucocyte and neutrophilcountsaswellasbloodSODlevelinter groupwereanalyzedusingindependentt-test.The different of measurement of each group was analyzedusingrepeatedAnova.Apvaluep<0.05 wasconsideredassignificant. RESULTS ThecharacteristicsofsubjectsofGuavajuice and Mineral water groups before exercise are presentedinTABLE1. Nosignificant difference was observed between Guava juice group and Mineralwatergroupbeforeexerciseindicatingthat bothofgroupsweresimilar.
56 J Med Sci, Volume 46, No. 2, June: 53 - 60 TABLE1. Characteristics of subjects of Guava juice and Mineral water groups The mean of BW, HR and body temperature ofsubjectsbeforeandafterexerciseinGuavajuice andMineralwatergroupsarepresentedinTABLE 2. Change in BW was observed before and after exercise in both groups (p<0.05). Heart rate of both groups before exercise was significantly different(p<0.05),howeveritwasnotsignificantly differetnafterwarmingup,exerciseandrecovery (p>0.05). The body temperature between Guava juice and Water mineral before exercise was significantly different (p<0.05), however after exerciseitwasnotsignificantlydifferent(p>0.05). TABLE2. Mean of BW, HR and body temperature of subjects before and after exercise in Guava juice and Mineral water groups ThenutrientintakeofsubjectsinGuavajuice and Mineral water groups during study are presentedinTABLE3.Asignificantlyhigherintake in energy, fat, protein, vitamin C, vitamin E, cuprum,zincandmanganwasobservedinGuava juice group compare to Mineral water group (p<0.05). However,the carbohydrate intake was not significantly different between two groups (p>0.05).
57 Ulvie et al., The leucocytes, neutrophils and superoxydes dismutase (SOD) level after consuming guava juice (Psidium guajava L) during aerobic exercise for beginners TABLE 3. Nutrient intake of subjects in Guava juice and Mineral water groups Theleucocytescountbeforeandafterexercise in Guava juice and Mineral water groups are presentedinTABLE4.Nosignificantlydifference in the leucocytes count before and after exercise in both groups was observed in this study (p>0.05). TABLE 4. Leucocytes count (µL -1 ) before and after exercise in Guava juice and Mineral water groups Thepercentageofneutrophilsbeforeandafter exerciseinGuavajuiceandMineralwatergroups are presented in TABLE 5. No significantly differenceinthepercentageofneutrophilsbefore exercise and after exercise on day 14 between two groups was observed (p>0.05). However, after exercise on day 1 and 7 the percentage of neutrophilsinGuavajuicegroupweresignificantly higherthanthat inMineralwatergroup(p<0.05). Duringtheexercise,thepercentageofneutrophils in Guava juice group significantly increased (p<0.05), whereas in Mineral water group was notsignificantlydifferent(p>0.05).
58 J Med Sci, Volume 46, No. 2, June: 53 - 60 TABLE5. Percentage of neutrophil (%) before and after exercise in Guava juice and Mineral water groups ThebloodSODlevelbeforeandafterexercise in Guava juice and Mineral water groups are presented inTABLE 6.Although the blood SOD level tendedto be higher after exercise, however nosignificantlydifferenceinthebloodSODlevel before and after exercise in both groups was observed in this study (p>0.05). TABLE6. Level of blood SOD (U/mL) before and after exercise in Guava juice and Mineral water groups DISCUSSION Regular physical exercise has many health benefitsincludingareducedriskofcertainsdiseses such as cardiovascular disease, cancer, and diabetes. 14,15 Otherwise,itisalsowidelyaccepted that contracting skeletal muscles generate free radicals and that prolonged and intense exercise canresultinoxidativestressanmuscleinjury. 1,6,7,16 Furthermore, the oxidative stress has substantial effect on leucocytes life span.The amounts of several hormones, cytokines and other factors whichmightinfluencecellularsurvivalareincreased or decreased in organs, tissues and peripheral bloodduring exercise. 17,18 ThisstudyshowedthatinMineralwatergroup the amount of leucocytes and percentage of neutrophil tended to increase after exercise on day 1 and 7, although it was not significantly different. It was indicated that actually aerobic exercise for beginnerinduce oxidativestress that lead to stimulate leucocytes production due to additionalunitmotorrecruitmentandmuscleinjury. However, ROS production due to the oxidative stress might be neutralized by endogenous antioxidantSODthatfoundalsotendedtoincrease inthisstudy.Althoughitwasalsonotsignificantly different.Resultofthisstudyisinaccordancewith study conducted by Shojaeiet al. 9 thatreported that leucocyte increased immediately after 45
59 Ulvie et al., The leucocytes, neutrophils and superoxydes dismutase (SOD) level after consuming guava juice (Psidium guajava L) during aerobic exercise for beginners minutes cycling with intensity 50% of VO 2 max foruntreatedhuman. Furthermore,inthisstudyalsofouundthatthe amountofleucocytesandpercentageofneutrophil tended to decrease after exercise on day 14, althoughit was not significantlydifferent. These decreasemightbecausedbyleucocytesapoptosis after the prolongation of aerobic exercise. Exercise-induced leucocyte apoptosis has been reportedpreviously.Syuetal. 19 demonstratedthat acute severe exercise induced anoxidative stress which resulted in acceleration of spontaneous neutrophil apoptosis. Moreover, repeated moderate exercise (30 min a day, 5 days a week at60%ofmaximalworkload)delayedneutrophil apoptosis. 20 Arecentstdyreportedthatasignificant delayofneutrophilapoptosis aftermarathonrun, intensiveenduranceanddownhillrunningaswell as intensive resistance exercise was observed. 21 SimilarresultswereobtainedonGuavajuice groupfor leucocytes which tendedincrease after exercise on day 1 and 7 and decrease on day 14, whereas SOD level tended to increase on day 7 and 14. Although, these were not significantly different. However, the percentage of neutrophil significantlyincreasedduringaerobicexercise.It wasindicatedthatguavajuicecanprotectexercise- inducednutrophilapoptosis. The effect of guava fruit to protect exercise- inducedneutrophil apoptosis might becaused its antioxidantactivity.Guavafruitisrichantioxidant such as vitamin C, vitamin E, lycopene and flavonoids. Guava fruit is also a good source of mineralssuchascopper,iron,zinc,magnesiumand selenium. 11-13 Moreover,theantioxidantactivityof guavafruithasbeenprovenbysomeauthors. 13,22,23 Antioxidant supplements may benefit exercise performance directly through the reduction of muscle fatigue at the level ofcontractile function andindirectlythroughreductionofphysiological stressors that negatively impact on training or improvement in the ability to recover from training. 6,24 CONCLUSION It can be concluded that consuming guava juice during exercise for the beginners does not signifincantlyinfluenceleucocytecountandSOD levels. However, the consuming guava juice can significantlyincreasepercentageofneutrophil. ACKNOWLEDGEMENTS We would like to thank all volunteers who participated in this study. We also grateful to Director of STPN,Yogyakarta Prof. Endriatmo Soetartowhohasgivenhispermissiontoconduct thisstudy.Thisstudycannotbeconductedwithout valuable assistances from technicians of Biochemistry Laboratory, Faculty of Medicine, UniversitasGadjahMada. REFERENCES 1.Penedo FJ and Dahn JR. Exercise and well-being: a review of mental and physical health benefits associated with physical activity. Curr Opin Psychiatry 2005; 18(2): 189–93. Doi:10.1097/ 00001504-200503000-00013 2.Puetz TW, O’Connor PJ, Dishman RK. Effects of chronic exercise on feelings of energy and fatigue: aquantitative synthesis. Psychol Bull 2006; 132(6): 866–76.Doi:10.1037/0033-2909.132.6.866 3.Foss LM. Physiological basis for exercise and sport. NewYork: McGraw Hill Book Company, 1998. 4.Len J, Davies CT, Young K.
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Description
The study investigates the impact of guava juice on oxidative stress during exercise.