2010) 1( 23مجلة ابن الھیثم للعلوم الصرفة والتطبیقیة المجلد في أمصال E دراسة مقارنة لمستویات الشد التأكسدي وأكسدة الدهون وفیتامین كبار السن المصابین بالفصام العظمي واألصحاء عابدة أسماعیل ابراهیم العلیوي جامعة بغداد ،ابن الهیثم –كلیة التربیة ،لكیمیاء قسم ا الخالصـة دلیًال على أنُه من أكثر (OA)أنتاج أصناف األوكسجین الفعالة في معظم األمراض ومن ضمنها الفصام العظمي ان لك بمقارنة المالون ثنائي وألجل التحري عن دور الجذور الحرة عند مرضى الفصام العظمي وذ .العملیات الضارة بخالیا الجسم M)األلدیهاید DA) على أكسدة الدهون ومستوى فیتامین دلیالE من مضادات األكسدة عالیة الكفاءة في امصال دالذي یع .المرضى مع مجموعة من األصحاء راذك 18 ( الذین تم تشخیصهم باستخدام تقنیات تشخیصیة مختلفة بأنفصام العظمي للركبة امریض 34شملت الدراسة أظهرت نتائج .) ثىإن 16و را ذك (16 من األصحاء 32كذلك شملت الدراسة سنة 78-45تتراوح أعمارهم بین ) ثىإن 16و مقارنة مع مجموعة )النساء والرجال(في مصل مجموعتي المرضى MDAالدراسة إن هناك زیادة معنویة في مستوى الـ لمجموعتي المرضى مقارنة مع مجامیع Eفي مستوى فیتامین امعنوی اوجد انخفاض كذلك .ة ألعمار المرضىبالسیطرة المقار .األصحاء IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (1) 2010 Comparative Study of Qxidative Stress, Lipid Peroxidation Marker, and Vit E Levels in Sera of Elderly Osteoarthritis Patients and Healthy Control. A. I. Al-Ela wy Departme nt of Chemistry,Colleg of Education Ibn-Al-Haitham , Unive rsity of Baghdad Abstract The production of reactive oxy gen sp ecies in most diseases including ost eoarthritis (OA) is confirmed to be the most destractive p rocess to human cells. To invest igate the role of free radicals in p atients with OA and by comp aring lip id p eroxidation marker maloan dialdehyde (M DA) and Vit E as one of the p otent antioxidant in Sera of (OA) p atients with those in healthy p op ulation. The st udy includes 34 p atients with knee OA diagnosed by doctors using different diagnost ic p arameters techniques , 18 females and 16 males and their age ranged 45 – 78 y ears , in addition to 32 healthy control 16 males and 16 females. The result revealed a si gnificant elevation in serum M DA sera in both p atient group s ( male and female) comp ared to match aged healthy control group s. On the other hand a si gnificant decrease in seru m Vit E lev els in both p atient groups compared to control group s, was not iced. Introduction The energetic benef it of aerobic metabolism is associated with gener ation of free r adicals (FRS).These radicals are high ly reactive molecules generated by the redo reaction that occur as p art of normal cell metabolism, further FRS may be formed by some reactive enzy mes[1]. Lip id p eroxidation is a well established M echanism of cellular injury in human, and is used as an indicator of o xidative st ress in cells and tissues. Lip id p eroxides, derived fro m p oly unsaturated fatty acids, are unst able and deco mpose to form a comp lex series of compounds. These include reactive carbony l comp ounds, which are the most abundant malondialdehy de (MAD) which is a highly reactive b ifunctional molecule, and has been shown to crosslink erythrocyte p hosp holip ids and p roteins. This p rocess results in imp airment of the membrane- related functions that ultimately leads to diminished surviv al. Therefor e, measurement of malondialdehy de is widely used as an end product and indicator of lip id peroxidation. Increased levels of lipid p eroxidation p roducts have b een associated with a variety of disease in humans (Voet and Vo et, 2004) [2]. On the other hand FRS could be generated by exp osure to environmental factors, also p roduce by activated phagocytes, and nurop hiles to kill p athogens[3]. The generation of reactive oxy gen sp ecies (ROS) via (FRS) formation are cap able of dama ging the biolo gically relevant molecu les such as lip ids, p roteins, carbohy drate, and DNA[4]. These processes may be involved in the initiation and p rop agation st ep s of several age related diseases[5]. Osteoarthritis (OA) is a degenerative joint disease. It results from the combination of genetic abnormalities and joint disease[6]. Among the multip le risk factors have been link ed to OA in epidemiology st udies age and IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (1) 2010 female versus male[7]. Aerobic cell have the cap acity to remove the active p roduct of oxy gen through endogenous cellu lar enzy matic defenses, additional cellular defenses include small antioxidant molecu les i.e. α-Tocop herol (vitamin E) is the most widely distributed antioxidant in n ature, when vitamin E donated an electron to a lip id p eroxy l radical it is converted to free radical st abilized by resonance structure.[8] The major function of Vit E in the body is an antioxidant and it acts p rimarily by scaven ging active o xy gen free radicals. It also p rotects other antioxidant from being o xidized. The capability is then also great in help ing to p revent the degener ative diseases[9]. The role of diet (comp osition of rich antio xidant sources food) in the treatment of OA remains unclear. Curr ent research into diet as an indep endent factor is focused on the merits of antioxidants (i.e. Vitamin)(Kalunian and Brion[10],( 2005) Since OA is by far the most common degenerative joint disorder throu ghout the world, and is one of the leading causes of disability and p ain in elderly , and the p revalence of this disease is less than 0.1% in those aged 25 y ears old v ersus a rate of over 60% in p eople over age 55 y ears[11]. So this st udy conducted on elderly p eop le more than middle aged. Subjects and Me thods The st udy was p erformed on (34) randomly selected p atients (18 females and 16 males) with OA of the Knee, at t he outpatient clinic of Baghdad Teachin g hosp ital in Baghdad-Iraq. The selected p atients have sy mptomatic and radio logic evidence of OA in one or both knee joints, and show different signs and sy mptoms. The control group includes 32 healthy p ersons (16 females and 16 males). 1 Blood samp les have been withdrawn from the st udy subjects by uenip uncture at 10am., in a disp osable tubes and centrifuged (250 xg for 10 min) within 45 min after collection to get t he serum which was st ored at -20 o C, unless used directly . 2 Determination of serum M DA:- M DA was measured according to[12]by monitoring the colored comp lex formed by the reaction of MDA with thiobarbituric acid the absorbance was t aken at 532 nm. 3 Determination of serum α-Tocop herol (vitamin E): High p erformance liquid chromatogr aphy (HPLc) technique, p rovided with UV detector was used[13]. The serum was deproteinized with ethanol; the vitamin and its ester were extracted by hexane, dried, and redissolved in chlorofor m p rior to injection on the silica column. The vitamin was eluted with 99% hexane and monitored at 287 nm. Ant ioxidant solution was used for the p resentation of α-Tocop heroloxidation during extraction; all steps were p erformed under st ream of nitrogen. Student -t-test was used to comp are the significan ce of d ifference in the mean values in comp arison group s , p<0.05 was considered st atically significant. Result and Discussion Table (1) showed the results of lip id p eroxidation mark er (MDA) in Sera of control group s and p atient with Ost eo arthritis resp ectively. No significant differences were found in serum M DA levels between male and female group s in healthy controls while a si gn ificant increase in serum M DA of male p atients comp ared to female p atients was noticed. From table (3)which showed significant differences b etween serum M DA levels of male patients (117.69±11.12) comp ared to serum level of control male IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (1) 2010 group s (35.3±4.3) also a si gnificant increase in f emales p atients (112.5±0.9) comp ared p atients group to that of healthy female (33.9±5.9). Oxidative damages are imp licated in most diseases, the level of M DA which is one of some p roduct of lip id peroxidation used as marker of free radicals oxidation of lip id. The effect of these radicals on jo ints of humans has been confirmed [14,15]. Tables (2) and (4) showed serum level of vit E in control group s, p atients with OA group s and serum level of vit E in st udied group s resp ectively. No significant difference between vit. E levels in female and male in control group s also a non significant decrease in female p atients comp ared to male p atients was found. A significant reduction in v it E levels in female p atients (0.32±0.03) compared to control (0.85±0.08) and a significant redu ction in Vit E of male patients (0.38±0.03) compared to male control group (0.85±0.08) was noticed. The reduction in Vit E in f emale and male group s comp ared to the healthy controls and the decline of this Vit may p romote the degener ative process in the p atient group s[16,17]. The increase in fr ee radicals generation is due to t he disease stage or the impartment of some antioxidant defenses and this a greed with other results rep orted that raise in (M DA) could be due to increase generation the active oxy gen sp ecies due to the excusive oxidative d amage generated in these patients. The oxy gen sp ecies in turn can o xidize many other important biomolecular including membranes l ipid [18]. Re ferences 1. Halliwell,B. and Gutteride,J.M . (1989) :Clarendon p ress : Oxford,UK. 2. Veot, D. and Voet, J.G. (2004): Biochemist ry ; Third edition, John and Sons, Inc. 3. Harp er, H.A., Radwel,V.W., and M ayes P.A., (2000) :″ Review of p hy siological chemistry ″ 17 th ed, M c Graw Hill. 4. Halliwell,B. (1994): Lancet 344(10) :721-734. 5. Bogdanska,J.J.; korneti, P. and Todorova, B. (2003) Nutrients 104(3):108-114. 6. Brandt, KD. (2005) Harrison’s p rinciples of Internal M edicine, 6 th Edition, Kasp er DL (Ed); M c Graw Hall comp anies., 2036-2045. 7. Brandt, K. (1997) :Text book of Rheumatology , (5 th ed). Kelley WN;Harris Jr.ED; Ruddy S; Sledge C E(Eds), WB. Saunders, p hiladelp hia.; 1303, Evidance for the p hotop rotective effects of vitamin E. 8. Baynes,J.W. (2005) Oxy gen and life in: Baynes, J.W., Domoniezak M .H. editors, M edical Biochemist ry . Philadelp hia: Elsevier ; p. 497-506. 9. Sokol, R.J.(1996) EE Zieglar and LIFler, editors, Washington D.C.Ls p ress 130-136. 10. Kalunian, Kc. ; Br ion, p H. and Conco, FF., (2005): Wollaston SJ: Non-p harmacologic therapy of Ost eoarthritis,UP to date. 11. Jordan, J. M .;Luta, G. and Renner, JB. (1996) : Arthritis care Res.;9:237. 12. Pesce, A.J. and Kap lan,.L.A. (1987) :″ M ethods in clinical chemistry ″ M osby comp any, London. 13. Farwer,S.R.; DerBo er,B.C.J.; Hadde man, E.; kivits,G.A.A.; wiersma, A and Danse, C. (1994): British J. Nutrition ; 72:127. 14. Green wald, R.A.; and M ay, W.W.,(1980): Arthritis; 23:455-463. 15. Sie, SH. (1997):Antioxidants in Disease M echanisms and therapy , London:Academic p ress. 16. Hermandez- Saavedr a D. and M c cord, J.M . ,(2003):Webb waring Inst itut e for cancer, University of Colorado Health Sciences center, Denver, co loradon USA. 17. Sidd hartha P. (2007): Indian Journal of M edical sciences; 61:9-14. 18. Kennett , E.C. and Kuchel, P.W, (2003) : IUBM B Life; 55: 375-385. IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (1) 2010 Table (1) MDA&Vitamin E levels of all studied groups Subjects No. MDA mg/dL P value Vit E mg/dL P value Te 32 34±5.1 0.850±0.075 Fe 16 33.9±5.9 Ns<0.05 0.850±0.075 Ns<0.05 M e 16 35.3±4.3 Ns<0.05 0.850±0.075 Ns<0.05 TOA 34 115.095 ± 11.0 0.35± 0.03 FOA 18 112.5± 10.9 S<0.05 0.32± 0.028 Ns< 0.05 MOA 16 117.69± 11.12 S<0.05 0.38± 0.031 Ns< 0.05 Not e: All Valu es shown in tables are mean ± SEM Te = Total healthy control. Fe = Female healthy control. M e = M ale healthy control. TOA = Total p atient with Knee Ost eoarthritis. FOA = Female with knee Osteoarthritis. MOA = M ale with Knee Ost eoarthritis. S = Sign ificant p . value. NS = Not significant p . value. Table (2) MDA & Vitamin E levels in sera of controls and patients groups. Subjects MDA mg/dL Vit E mg/dl P values Fe 33.9±5.9 0.85± 0.08 S<0.05 M e 35.3±4.3 0.85± 0.08 S<0.05 FOA 112± 0.9 0.32± 0.03 S<0.05 MOA 117.69± 11.12 0.38± 0.03 S<0.05