Microsoft Word - 15 493 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 Evaluation of IL-35 and hs-CRP in Iraqi Patients with Cardiovascular diseases Warka'a T. Al-Sa'adi Dept. of Chemistry/College of Education for Pure Science ( Ibn Al- Haitham)/ University of Baghdad Received in: 22 June 2013 ,Accepted in:28 October 2014 Abstract Interleukin-35 (IL-35) is a novel anti-inflammatory of IL-12 family member cytokine. High sensitive C- reactive protein (hs-CRP) is a marker of inflammation disease. This study was obtained to estimate the concentrations of IL-35 and hs-CRP in cardiovascular diseases Iraqi patients. The study contained 79 subjects divided into two groups, 47 (21 male and 26 female) suffering from cardiovascular diseases and 32 as control group. The concentration of IL-35 and hs-CRP was measured by enzyme- linked immune sorbent assay (ELISA). The results revealed that the concentrations of IL-35 and hs-CRP are increased in patients group. IL-35 is strongly expressed in human advanced plaque and hs-CRP. Therefore these two biomarkers could be useful as addition in diagnosis of cardiovascular diseases. A significant increase of IL-35 and hs-CRP levels in patients group compared with controls (P<0.005) and there was a positive correlation between IL-35 with hs-CRP in patients group. The results of this study revealed that IL-35 is a potentially anti-inflammatory marker in cardiovascular diseases. Key Words: Inflammatory markers, IL-35, hs-CRP, Cardiovascular diseases, Iraqi patients. 494 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 Introduction Inflammatory plays an important role in the pathogenesis of cardiovascular disease and various studies in dicated that inflammatory markers appear to be protein of cardiovascular events such as interleukin-6, IL-10 and C-Reactive protein (CRP)[1-5]. Interleukin 35 (IL-35) is the newly identified member of IL-12 family[6,7]. IL-35 is a heterodimeric cytokine composed of p35 subunit of IL-12α and the Epstein-Bar Virus induced gene3 (EBI3)[7,8]. EBI3 is expressed at high levels in human B lymphoblast cells, tonsils and spleen [9]. IL-35 is secreted by CD4+ regulatory T (Treg) cells rather than CD4+ effector T (Teff) cells in mice; therefore, IL-35 has been considered to be the characteristic factor of Treg cells[10–13]. Human Treg cells, placental trophoblast cells, endothelial celss, activated dendritic cells, and macrophages also express IL-35[14,15]. Additional evidence demonstrated that IL-35 is an important anti-inflammatory cytokine and can efficiently suppress the Teff cell (including Th1, Th2 and Th17 ) activity and reduce the progression of inflammatory diseases and autoimmune diseases [6,7,16,17]. C-reactive protein (CRP) is the prototype acute phase protein primarily synthesized in liver and its release is stimulated by interleukin6 (IL-6) and other pro inflammatory cytokines[18]. It is composed of five identical non-glycosylated polypeptide subunits each containing 206 amino acid residues. CRP is a hepatically derived pentraxin that plays a key role in the innate immune response[19,20]. Epidemiological studies have demonstrated that C-reactive protein (CRP) is a strong predictor myocardial infarction [21-23], stroke[21,22,24] sudden cardiac death[25], and peripheral arterial disease[24] in apparently healthy adults. High sensitive CRP (hs-CRP) is a marker of low grade chronic system inflammation and it can amplify the anti-inflammatory response through complement activation, tissue damage, and activation of endothelial cells [26-28]. There is no information mentioned to the relationship between the levels of plasma IL-35 and its potential role in cardiovascular disease for Iraqi patients. This present study was carried out to assess the levels of serum IL-35 in heart diseases patients and the relationship to other parameters, including hs-CRP and total cholesterol (TC). Methods The study consisted of 79 subjects divided into two groups, 47 subjects (21male and 26 female, range age 48-68 years) suffering from Cardiovascular diseases. The other 32 health were controlled as control group (male 13 and 19 female, range age 44- 65 years). The patients were taken from CCU unit in AL-Kadymia teaching Hospital for period from January to April, while the healthy controls were volunteers. Blood samples (5 ml) were drawn from patients and controls. The levels of serum IL-35 and hs-CRP were measured by using an enzyme- linked immune sorbent assay (ELISA), according to the manufacturer’s instructions (CUSABBIO) and (DRG) respectively. Serum total cholesterol (TC) was estimated by commercially available kit from BioSystems. It was estimated by enzymatic method. Statistical Analysis All data are given as the mean ± SD and where analyzed by an ANOVA. Person’s correlation was used to examine the relation between IL-35 with hs-CRP and TC. A value of P was considered to be statically significant (P< 0.005). 495 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 Results The distribution of sex, mean age and total cholesterol for patients and control groups are shown in table 1. The mean age was (50± 4.3) years in Iraqi patients and (56± 6.9) in control group. There was no significant difference in age and gender between the Iraqi patients with cardiovascular diseases and control group as in Table 1. Serum total cholesterol showed significant differences between patients and control groups (P < 0.005). Mean value of serum total cholesterol (TC) was 228.6 ± 21.05 mg/ dl in patients group while in control group it was 175.34 ± 12.05 mg/ dl. Table 2 showed mean value of serum IL-35 for patients and healthy control groups which it was 36.21 ± 3.52 pg/ dl and 22.21 ± 2.01 pg/ dl respectively. Mean values of hs-CRP were 4.36 ± 0.76 mg/ L and 0.77 ± 0.25 mg/L for patients and control groups respectively. Discussion The levels of anti-inflammatory cytokine IL-35 were measured in cardiovascular diseases for Iraqi patients. The results showed that the elevated concentration of IL-35 was associated with cardiovascular diseases and positively correlation. The levels of anti- inflammatory cytokine IL-35, hs-CRP and total cholesterol were investigated in cardiovascular diseases for Iraqi patients. The results showed that the levels of IL-35 and hs-CRP were significantly higher in patients than in control group. The results showed that higher IL-35 levels were positively correlated with CRP (R= 0.11), (figure 3) whereas higher TC levels were negatively correlated with IL-35 (R= -0.1) in patients with cardiovascular diseases (figure 4). The serum anti-inflammatory cytokine concentrations levels are still controversial. Some studies observed that these levels were increased other studies observed significantly decreased[10-18,29,30]. These studies indicate that the concentrations of anti-inflammatory cytokines can predicate the incidence of cardiovascular events. IL-35 is an IL-12 family member cytokine composed of an  chain p35 and a  chain EBI3[6,7]. EBI3 and p35 are expressed in almost all advanced plaque lesions and are co- expressed in atheroma vascular smooth muscle cell, indicating that IL-35 may be secreted by vascular smooth cells and is involved in the atherosclerosis[31- 34]. In addition, higher IL-35 concentrations showed a positive correlation while higher TC concentration showed a negative correlation with IL-35 in patients with cardiovascular diseases, suggestion a potential role of IL-35 in prognosis of cardiovascular diseases. It was found that elevated hs-CRP was significantly correlated with coronary artery disease[33,34]. A significant difference has been observed regarding the value of hs-CRP in cardiovascular disease subjects compared to normal subjects in this study. This reveals that there is as association between hs-CRP and cardiovascular diseases[27]. 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PLoS ONE 9(1): e87787. 498 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 33- Vahdat, K.; Jafari SM.; Pazoki R. and Nabipour I. (2007). Concurrent increased hs-CRP and chronic infection are associated with cad, a large population study. Indian Journal of Medical Sciences 61: 135-143. 34- Pasupathi ,P.; Rao YY.; Farook J.; Saravanan G. and Bakthavathsalam G.(2009). Oxidative stress and cardiac biomarkers in patients with acute myocardial infarction. European Journal of Scientific Research 27: 275-285. Table No.(1): Clinical characteristics of Iraqi patients and control groups Characteristics Patients n=47 Control n= 32 P value Age (years) 50± 4.3 56± 6.9 N.S Sex (male/ female) 21/26 13/19 N.S TC (mg/ dL) 228.61± 21.05 175.34± 12.05 P < 0.005 The data are given as the mean ±SD. TC: total cholesterol, N.S: nonsignificant. Table No.(2): plasma IL-35 levels and hs-CRP in patients and control group Characteristics Patients Control P value IL-35 (pg/dl) 36.21± 3.52 22.21± 2.01 P < 0.005 hs-CRP (mg/l) 4.36± 0.76 0.77± 0.25 P < 0.005 Note: The data are given as the mean ±SD 499 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 Figure No. (1): The correlation between IL-35 and hs-CRP in patients groups Figure No. (2): The correlation between IL-35 and TC in patients groups. 500 | Chemistry 2014) عام 3(العدد 27المجلد مجلة إبن الھيثم للعلوم الصرفة و التطبيقية Ibn Al-Haitham Jour. for Pure & Appl. Sci. Vol. 27 (3) 2014 وبروتين سي عالي الحساسية للمرضى العراقيين المصابين 35-تقدير انترلوكين بأمراض األوعية القلبية وركــاء طعمة سلوم السـعدي كلية التربية للعلوم الصرفة (ابن الھيثم)/ جامعة بغداد /قسم الكيمياء 2014تشرين االول 28,قبل البحث في: 2014حزيران 22استلم البحث في: الخالصة ضد األلتھابات. بروتين سي عالي الحساسية ھو مؤشر 12-عضو جديد من عائلة انترلوكين 35- انترلوكين وبروتين سي عالي الحساسية عند المرضى العراقيين 35-أجريت ھذه الدراسة تقدير تركيز انترلوكين لأللتھابات. انثى) 26ذكراُ و 21حالة ( 47حالة، قسمت على مجموعتين: 79المصابين بامراض األوعية القلبية. تضمنت الدراسة يطرة. حالة غير مرضية مجموعة الس 32يعانون من امراض األوعية القلبية و 35-وبروتين سي عالي الحساسية بطريقة اإلياليزا. اظھرت النتائج زيادة في تركيز انترلوكين 35- قيس تركيز انترلوكين وبروتين سي عالي الحساسية يُعبر عند األشخاص 35-وبروتين سي عالي الحساسية في مجموعة المرضى. انترلوكين ن ھذان المؤشران الحياتيان مفيدين في تشخيص امراض األوعية القلبية. بشدة في الحاالت المتقدمة، لذا يمكن أن يكو وبروتين سي عالي الحساسية في مجموعة المرضى مقارنة بمجموعة 35- حصلت زيادة معنوية في مستوى انترلوكين . نتائج وبروتين سي عالي الحساسية لمجموعة المرضى 35-وھناك عالقة موجبة بين انترلوكين (P< 0.005)السيطرة مؤشر محتمل ضد األلتھابات في امراض األوعية القلبية. 35-الدراسة أظھرت انترلوكين ، بروتين سي عالي الحساسية، امراض األوعية القلبية.35- داالت األلتھابات، انترلوكين الكلمات المفتاحية: