IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Evaluation Interleukin ─ 8 and High Sensitivity C - Reactive Protein in Sera of Control Group and Human Female Patients with Osteoarthritis Nodal in Iraqi's Patients B. H. AL-Wihaly Department of Chemistry ,College of Education Ibn – Alhaitham, University of Baghdad Received in : 13 March 2011 Accepted in : 30 May 2011 Abstract The present study conducted on 30 female patients with osteoarthritis OA a attending Baghdad teaching hospital, in addition to 30 healthy females, all subjects were (35-65) year's old .Parameters were measured in the sera of patients group and healthy groups were interleukin -8 (IL-8) and high sensitivity C-reactive protein (hs-CRP).The results revealed a higher significant in all parameters of patients sera compared to healthy groups. Conclusion was obtained that IL-8 and hs- CRP were a markers or player inflammatory disease like osteoarthritis nodal Key words: Interleukin -8(IL-8),High sensitivity CRP(hs-CRP) ,Osteoarthritis Introduction Osteoarthritis (OA) is a slow progressive disorder of synovial joints .This joints disorder is characterized by a loss of balance between synthesis and degradation of the articular cartilage constituents leading to subsequent erosion of joint cartilage remodeling of the underlying bone oseophyte formation and variable degree of synopitis [1].When clinical characteristics of OA (pain loss of mobility and radiographic narrowing of the joint space ) manifest , the actual changes in articular cartilage and subchondrla ,bone have started long ago[2]. Interleukins are group of cytokines [3] because they regulate in formation transfer among different types of leukocytes during various stage of immune or inflammatory response [4] secreted Proteins/signaling molecules/ that were first seen to be expressed by white blood cells (leukocytes). The term interleukin derives from (inter-) as a means of communication '' and (-leukin)'' deriving from the fact that many of these proteins are produced by leukocytes and act on leukocytes, it has since been found that interleukins are produced by a wide variety of body cells . The function of the immune system depends in a large part on interleukins and rare deficiencies of number of them have been described, all featuring autoimmune diseases or immune deficiency, the majority of interleukins are synthesized by helper CD4+T lymphocytes as well as through monocots macrophages, and endothelial cells. They promote the development and differentia on of T, B, and hematopoietic cells. Interleukin – 8 one of these interleukins secreted from macrophages, lymphocytes, epithelial cells and endothelial IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 cells, CXCR1/IL8RA, CXCR2/IL8RB/CD128 were the target receptors, Neutrophiles, lymphocytes were target cell [3] so as IL-8 was chemotaxis for Neutrophils [5]. C-Reactive protein (CRP) is member of the class of acute-phase reactant as its levels rise dramatically during inflammatory processes occurring in the body [6, 7][atherosclerotic plaques][9] .CRP binds to phospocholine on microbes, it is thought to assist in complement binding to foreign and damaged cells and enhances phagocytosis by macrophages (opsonin mediated phagocytosis) ,which express a receptor for CRP[10]. It is also believed to play another important role in innate immunity as an early defense system against infections [11] so is used as a clinical marker of a cute systemic inflammation and infection [9] CRP a rise up to 50,000 fold in acute inflammation such as infection .It rises above normal limits within 6 hours, and peaks at 48 hours [12] . A high – sensitivity CRP (hs-CRP) test measures low levels of CRP using laser nephelomoetry the test gives results in 30 minutes with sensitivity down to 0.04 mg\ L. Normal concentration in healthy human serum is usually lower than l0 mg \L, Slightly increasing with ageing, higher levels are found in late pregnant women, mild inflammation and viral infection (10-40 mg \L), severe bacterial in fections and burns 200 mg \L [13] Aim of the Study The aim of this study is to evaluate some biochemical parameters such as interleukin - 8, (IL-8), hs-CRP in sera of OA female patients which is used as a clinical marker of an acute inflammation and infection. Experimentaly Subjects The present study was performed on a group of 30 human females aged (35-65) years with on diagnosed by examining the patients by in the (Baghdad teaching) hosp ital and by x- ray examination on. In addition, a group of 30 healthy females were enrolled in the study as control group. Sampling Blood samples of 5ml were drawn all from subjects enrolled in this study, and kept in plain tubes 1eft to clot at room temperature for /5 min. then centrifuged at 3500 g for 10 min to separate the serum. - Interleukin – 8 (IL-8) determinations Interleukin 8(IL-8) has been estimated by using enzyme immunoassay (ELISA) technique. The procedure was done according to the manufacturer instruction as supplied with kit from Biosource Europe S.A. Company, Belgium. - High sensitivity C - reactive protein (hs CRP) determination - High sensitivity C - reactive protein (hs CRP) enzyme immunoassay for the quantitative determination in human serum was used. DRG international. Inc. USA. This is done by ELASA test. Statistical analysis The results were expressed as mean± SD of mean, using student-test, significant Variation is considered significant when p-values are ≤ 0.05. Results and Discussion Results obtained in the present study state that the serum level of IL – 8 was significantly higher in females patients with OA than in healthy control. (Table1),figure(1). The elevated IL-8 level found in the serum of females' patients with OA of the present study is in agreement as in pervious studies [14, 15] who found increase in the level of IL-8 in serum of OA patients compared with control and found that serum levels of IL-6 in arthritis patients [like fibro myologi a syndrome (FM S) patients] was the same as in controls. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 IL -8 is the major chemoattractant for neutrophils; it mediates changes in neutrophil shape. Transendothelial migration, induces exocytosis of granule enzymes such as elastase and myeloperoxidase and a resp iratory burst where hydrogen peroxide and super oxide radical are produced [15] IL-8 was upregulated by proinflammatory signals such as IL-1 and IL-17 and inhibited by anti -inflammatory cytokine (IL-4, IL-10) and glucocorticoids [16] furthermore IL-8 has been defined as a mediator of sympathetic pain [17]. C - reactive protein Results obtained showed that the serum level of CRP was significantly higher in patients with OA than in controls (Table1),figure(2). These results are in agreement with other studies reported that CRP level is regularly used in the clinic, which is useful to discuss it in terms of a diagnostic tool and therefore as indicator of an underlying disease in acute infections.[18] Less in known on the CRP level in chronic disorders that have an Inflammatory disease component [19] High sensitivity C-Reactive protein activates the complementy system with a primary function to bind and detoxify endogenous toxic substance produced as a result of tissue damage [20] CRP is not only a marker of, but a player in, cardiovascular disease (CVD)., CRP may directly promote atherosclerosis and endothelial inflammation by attenuating the release of NO, a key molecule in the endothelium that p lays a pivotal role in the maintenance of vascular tone [21] In addition, besides the potent vasodilator effects of NO, it meditates many protective functions in the endothelium, for example inhibiting the of pro-inflammatory cytokines, chemokines and leukocyte adhesion molecules, there by limiting vascular recruitment of leucocytes and platelets [22].NO also inhibits vascular smooth muscle cell proliferation, an early sign of atherosclerosis. Therefore CRP may have an important deleterious role in the atherosclerotic process. i .e .by inhibiting NO formations however the involvement of CRP as a functional player in the endothelium has recently been questioned as it has been suggested that the pro-inflammatory effects by CRP might be due to contaminating artifact in the commercial CRP preparation [23]. Over the last few years there have been a lot of p romising clinical markers proposed to link inflammation and atherosclerosis. One of these markers, hs – CRP [24].And these studies are in agreement with this study. From the present study, a conclusion could be stated that IL-8 increase concentration in sera of OA patients is due to interleukins that, regulate information transfer among different types of leukocytes during various stage of immune or inflammatory response, which is confirmed by increase in hs –CRP concentration. This finding may indicate that inflammatory activity might influence cytokine production in these patients [25] [26] References 1-Ana M.Valdes and Timothy , D. Specttro (2009) The contribution of Genes to osteoarthritis 66-(1):4593J.Medical Clinical of North America 2-Robert, B.Wallace; Johan, M. Last, et al(2008) "Public Health & Preventive Medicine " 50 th Edition ,McGraw .Hill Companies ,Inc(U.S.A) p .1128-1129 3- Lippincott Williams and Wilkins (2007) :Lippincott's Illustrated Reviews Immunology. 3 th Edition Lippincott Williams Wilkins ,U.S.A Page 68. 4- Devlin M.Thomas: cytokine Receptors (2006): text book of Biochemistry with clinical correlations, 6 th Ed. A John Wiley &Sons, Inc. , USA, p.507. 5- Karen M.Henkelsa., et, al. (2011). IL-8-induced neutrophil chemotaxis is mediated by Janus kinase 3 (JAK3). Edited by Beat Imhof 585(1): 159-166. 6- Danesh ,J. ;wheeler ,JG.; Hirschfield ,G.M . et al (2004) C-reactive protein and other circulating markers of inflammation in the prediction of coronary heart disease .N. Engl 1397 -: 1387 350.J. Med. (3): 74 30reactive protein, J. Australian prescriber -R. (2007) C,Gleen -7 8- Harper's (2006) Harpers lustrated Biochemistry 27 th ed. Lange medical Books McGraw- Hill Medical publishing Division NewYork p590- 591. 9- Päiväniemi O.E. paulak, et al (2009): Local C-Reactive protein Expression in obliterative lesions and the Bronchial wall in post transplant obliterative Bronchiolitis Volume 2009(2009), Article ID 510254, 7 pages doi : 10. 1155/2009/510254. 10- Pepys M.B. Hawkins p.n., et al. (2005) :proinflammatory effects of bacterial recombinant human C-reactive protein are caused by contamination with bacterial products , 103.-:97 97reactive protein itself .Circ.Res. -not by C 11- Pickup J.C, (2004): Inflammation and activated innate immunity in the pathogenesis 823.-:81327of type 2 diabetes .Diabetes care 12- Ron Kennedy.M.D.(2011) C-reactive Protein (CRP), Cardiac C-reactive Protein, (High Sensitivity C-reactive Potein) .The Doctors' Medical Library. 13- Castoldi G., Galimberti S., et al .(2007): Association between serum values of C-reactive protein and cytokine production in whole blood of patients with type 2 diabetes .Clin . Sci. 108 -: 103113 14- Gur A, and Karakoc M.(2002) Cytokines and depression in cases with 61. -(2): 35829fibromyalgia .J. Rheumatol. 15- Ransohoff RM, and Benvenistc EN.2005 Cytokines and the CNS .2 th ed .London, New York: Taylor &Francis Boca Raton ; pp217. 16- Uceyler N, Valenza R, and Stock M.2006. Reduced levels of anti- inflammatory cytokines in patiente with chronic widespread pain : Arthritis Rheum ; 54 : 2656- 2664 . 17- Meyer TD , Hautninger M, Allgmine . (2003) .Depressions Skala Normuerung an Minderjahrigen and Erweiterung zur Erfassung manischer symptom .Blood 365.-:360132mononuclear cells. Clin EXP Immunol. 18-Salzman B, .E. (2007). A primary care approach to the use and interpretation of common rheumatologic tests. Clin Fam Pract, 335-358. 19- Marnell L,Mold C, DU Closs TW .(2005) C-reactive protein: ligands, recpcors and role 11.-:104117 in inflammation .Clin Immunol; 20- Devaraj S, and Du closs TW , Jialal I.(2005): Binding and internalization of C- Reactive protein by FC gamma receptors on human aortic endothelial cells 3.136-:135925mediates biological effects . Arerioscler thromb vasc Biol. 21- Verma, S., Wang, C, H., Li, et al. (2002). Aself - fulfilling prophecy : C-reactive protein attenuates nitric oxide production and inhibits angiogenesis. Circulation 919.-,913106 22- Verma , S., Kuliszewski , M. A., Li, S.H. et al (2004)C- reactive protein attenuates endothelial progenitor cell survival , differentiation and function : further evidence of a mechanistic link between C- reactive protein and cardiovascular 2067.-,2058109isease .Circulation d 23-pepys, M.B., Hirshfield,G.M .,Tannent ,G.A.Et. al. (2006) Targeting 1221 -: 1217440reactive protein for the treatmeent of cardiovascular disease. Nature -C 24- Szmitko, P.E., Wang, C.H., Weisel, R.D., de Almeida, J.R., Anderson ,T .J. and Verma ,S.(2003) New markers of inflammation and endothelial cell activation I. 1923. -,1917108Circulation -:79113Clinical .Science . reactive protein: amarker or player? -Thomas N. (2007): C -25 81. 26- Gaede, P., Ved , P., Larsen, N., Jensen, G.V., Parving, H.H. and Pedersen , O. (2003)Multifactorial intervention and cardiovascular disease in patients with type 2 393. -:383348diabetes N. Engl .J. M ed . IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Table (1): IL-8 and hs –CRP concentration in sera of OA patients and control P value Control N=30 OA patients n= 30 p<0.05 84.91±14.58 120.66±37.42 IL-8 Pg/ml Mean ±SD p<0.05 0.583±0.141 1.211±0.524 Hs-CRP mg/dl Mean± SD Fig .(1) : IL-8 concentration in sera of OA patients and control. Fig .(2): ) hs –CRP concentration in sera of OA patients and control. 2011) 3( 24المجلد مجلة ابن الھیثم للعلوم الصرفة والتطبیقیة في مصل الحساسیةتقدیر االنترلیوكین الثامن وبروتین سي الفعال العالي مریضات بالتهاب المفاصل العقدي في العراقمجموعة نساء أصحاء يالو حیلبشرى حمید جامعة بغداد ،كلیة التربیة ابن الهیثم ،قسم الكیمیاء 2011آذار 13:استلم البحث في 2011آیار 30:حث في قبل الب الخالصة مـن 30 آل عـنضـفمن مستشفى بغـداد التعلیمـي أخذتمریضة بالتهاب المفاصل العقدي 30تضمنت هذه الدراسة ة فـي مصـل، ةسـن 35-65المجمـوعتین بـین أعمـاروتراوحت .األصحاءالنساء ،الـدم للمجمـوعتین وقیسـت المتغیـرات الحیویـ إحصـائي معنـوي محسـوس مرتفـع فـرق میـع النتـائج جعكست . )الحساسیةوبروتین سي الفعال العالي ،ثامناالنترلیوكین ال( مثـل ،االلتهابیـة اإلمـراضلمعرفـة اهذین المتغیرین مقیاس المعنستنتج من ذلك انه یمكن است .المجموعتین في مصل نساء التهاب المفاصل العقدي