IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (2) 2011 Studies o n the Relationship Between Chromium(III) ion and Thyroid Peroxidase Activity in Sera of Patients with Thyroid Dysfunctio n H. G.Hasan, T. J.Mahmood, P. A. Ismael Departme nt of Chemistry ,College of Education, I bin-alhaitham, Unive rsity of Baghdad College of Medicine, Halwer Medical Unive rsity Departme nt of Chemistry ,College of Education Science, Salahuddin Unive rsity Received in :27 October 2010 Accepte d in :8 February 2011 Abstract The objective of this st udy was to determine the concentration of trace element chromium(III) and thy roid p eroxidase activity in human serum , and to find a relationship between the concentration of chromium(III) and thy roid p eroxidase activity in serum of p atients with hy p othy roidism, hy p erthy roidism and healthy subjects. Serum thy roid p eroxidase was measured by enzy me linked radioimmunoassay(ELISA) method and chromium determination was by atomic absorp tion sp ectrop hotometer .Comparing the values of chromium concentration and thy roid peroxidase activity in both samples showed that there were significant p ositive correlations between chromium levels and thy roid p eroxidase activity (P<0.01, r=0.11). The results showed that Serum thy roid p eroxidase activity and chromium levels were significantly higher in hy p erthy roidism p atients (152.0±4.6 IU/L) (2.159±0.15 p p b) resp ectively than normal (51.0±1.8 IU/L) (0.378±0.024 p p b) resp ectively and lower than normal in hy p othy roidism p atients (35.0±0.31 IU/L)(0.099±0.011 p p b) resp ectively. Key word : Chromium(III), Thy roid peroxidase, thy roid dy sfunction Introduction The biosy nthesis of thy roid hormone from thy roglobulin is cataly zed by thy roid p eroxidase (TPO), an integral membrane p rotein. TPO is also a major autoantigen in autoimmune thy roid diseases [1]. Large quantities of p urified TPO are essential for elucidating its st ructure and understanding its role in diseases activity [2]. Failure of organification could result from deficient thy roglobulin accep ter, p eroxidase enzy me defect and lack of H2O2. Altaration in the p rocess of iodide organification has been detected in almost all thy roid diseases [3]. The qualitative abnormalities in the enzy me TPO sy st em have been p roved in p atients with congenital thy roid diseases, and quantitative disturbances were observed in multinodular goiter, toxic adenoma, thy roiditis, and carcinoma [4]. Trace elements are known to influence hormones at levels of action, including hormone secretion and activity and binding to target tissue [5]. Conversely , hormones influence trace element metabolism at several levels of action, including excretion and transp orts of trace metals [6,7,8]. Hence trace elements assay in biological fluid can be used as diagnost ic or p rognost ic aid in p atients with different hormonal disturbance alongside with other biochemical p arameters[9]. Chromium(III) is an essential trace element for human. Chromium(III) is an ion needed for nutritional sup p ort for the thy roid gland to help insulin through reducing body IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (2) 2011 weight since insulin blocks p hosp hory lation and therefore op p oses the action of epinep hrine and can imp ede thy roid hormone p roduction [10,11]. Chromium(III) also help s to control cholesterol levels that oft en elevate in hy p othy roidism p atients as well as control blood sugar levels which are challenged by p oor adrenal and thy roid gland health [12]. Serum chromium concentration are likely to be increased above the reference range in p atients with metallic joint p rost hesis, longevity , increased lip id quantity , imp aired glucose metabolism, hy p erglycemia, and glycosuria [13,14]. The aim of this st udy was conducted to verify the relation of the thy roid dysfunction and TPO activity together with Cr (III) ion. M aterial and M ethod Subjects 92 p atients with hy p erthy roidism and 82 p atients with hy p othy roidism for both sexes were involved in the st udy they were 27-68 y ears of age. 92 healthy p erson for both sexes aged 19-54 y ears were p articip ated in this study and rep resented the control group . S amples Collection and preparation Volume of five milliliters of venous blood from patients and healthy subjects were drawn by utilizing disp osable p last ic sy ringes and transferred into st erile test tube. The blood was allowed to clot and centrifuged at 4000 rp m for 10 minutes. Sera were separated and stored at 20 Cº until analysis . Esti mation of trace element chromium Serum chromium was determined using Flame At omic Absorp tion Sp ectrop hotometer .Standards. Samp les and blanks for estimation of chromium were asp irated into a ( Perkin Elmer 6000) atomic absorp tion sp ectrop hotometer utilizing along-p ath air/acety lene burner and cathode lamp for chromium metal. Each sample was read three times at 1 second. The concentration of chromium was found by st andard addition method[15,16]. Esti mation of thyroid peroxidase (TPO) activity Serum thy roid p eroxidase activity was measured by Enzy me Linked Immunosorbent Assay (ELISA) using ORGENTEC thy roid p eroxidase (TPO) (ELISA) kit. The activity of TPO was assayed from calibration curve by interp olation . Results and Discussion The results and comp arisons of serum chromium levels and TPO activity between hy p erthy roidism, hy p othy roidism and control group s were demonst rated in Table (1,2).There was p ositive correlation between serum chromium levels and TPO activity . The results of the p resent st udy revealed that serum chromium levels of hy p othy roidism p atients were significantly lower (0.099±0.011 p p b) than the levels in normal subjects (0.378±0.024 p p b) as shown in Table (1). A significant increase in serum chromium levels was demonst rated in hy p erthy roidism p atients (2.159±0.15 p p b) as comp ared with that of normal subjects. Chromium is an essential mineral that is required in the maintenance of our health and it is essential to the metabolism of lip ids, p roteins, carbohy drates and insulin regulation [17,18]. Thy roid activity may directly or indirectly affect chromium st atus, low thy roid function may allow increased insulin secretion resulting in chromium loss or increased insulin p roduction sup p ress t hy roid function[19,20]. In this st udy significant decrease in the levels of Cr in hy p othy roidism p atients were observed .One p ossible exp lanation for these finding, that gastrointest inal absorp tion of chromium is severely imp aired in hy p othy roidism subjects [21,22,23,24]. Anot her exp lanation is due to the significant influence of TSH in the variation of the concentration of chromium in normal and altered human thy roid tissue [25]. High serum chromium levels have been reported in hy p erthy roidism and serum chromium concentrations have been noted to correlate well with thy roid gland activity [26]. Data obtained showed an increased serum chromium levels in p atients with hy p erthy roidism and positively correlation between serum chromium levels and TPO activity . Changes in body metabolic rate have been shown to be reflected in altered chromium metabolism [27]. IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (2) 2011 The increase in serum chromium levels in hyp erthy roidism p atients is in agreement with the st udies of other researchers indicating the imp ortant role of chromium in controlling the thy roid gland function [28,29,30]. Goncharo and Ametov [31], st udied the effect of chromium sup p lementation on the function of thy roid gland in exp eriment on albino male rats. The results of t heir st udy illustrated that chromium sup p lementation (3M g/body weight) imp rove thy roid activity in albino rats . Table (2), showed that the TPO activity in hy p othy roidism p atients was significantly decreased (35±0.31 IU/L), p robably due to a destruction that could occurs in thy roid tissue and rep lacement with fibroblast and ly mphocyte [32]. It has been reported that serum TPO activity decreases in patients with hy p othy roidism [33]. In hashimotos t hy roiditis a variable degree of infiltration with lymphocytes and fibroblalst s is commonly seen in the thy roid gland, therefore, the activity of tissue enzy me is p robably affected by the degree of tissue damage. The slight decrease of enzy me activity in the thy roid gland is somewhat intriguing considering the degree of tissue destruction which caused hyp othy roidism [34] . Defects of TPO are both quantitative and qualitative the alter include imp aired binding to heme, imp aired binding to thy rogobuline or iodine substrate, abnormal localization in the erythrocyte, and abnormal suscep tibility to inhibition [35]. Thy roid p eroxidase gene mutations have been identified causing either abnormal thy roid p eroxidase or absent enzy matic activity or comp lete absence of TPO p rotein formation [36]. As shown in table (1), TPO activity in hy p erthy roidism p atients was significantly elevated (152±4.6 IU/L) in comp arison with the normal subjects (51±1.8 IU/L). One p ossible exp lanation for these finding , that chronic TSH st imulation leads to increased iodide binding because of increased gland p eroxidase content [37], increased iodide trapp ing , and p resumably increased H2O2 generation [38]. hence increase TPO activity [39]. Anot her exp lanation is due to the significant influence of TSH in the concentration of iodine and chromium in normal and altered thy roid tissue. Thy roid p eroxidase activity was grossly elevated in toxic goiter. This may be of significance in the p athogenesis of graves diseases [40]. The relation between TPO activity and serum chromium levels is shown in Figure (2). A significant p ositive correlation was obtained between TPO activity and serum chromium levels in hy p erthy roidism and hy p othy roidism (r=0.11), when chromium levels were decreased by more than (70 %) the TPO activity was markedly decreased . Statist ically significant correlation were found among indexes of chromium st atus and indexes of TPO. Concomitants deficiency of both key p arameters are esp ecially dangerous. In other st udies, using animals chromium deficiency in rats inhibited the p roduction of T3 and T4 [41]. Anot her st udies revealed that chromium deficiency in animals can exert both a direct effect on the metabolic p rocess and an indirect one disturbing iodine metabolism. The results of this st udy indicated that chromium deficiency in animals enhances the effect of hy p othy roidism [42]. As shown in figure (3), from a tot al of 260 p atients with thy roid diseases the most common genders are females. It can be seen from figure (2), that 92 p ersons were male and 170 were females. Hi gh lev el of chromium was found in females. Our results are consistent with other studies such as who found by Lien et al [43], that t hey p roved thy roid disease affects female three times more than male . 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(2007) Bioelement effects on thy roid gland in children living iniodine-adequate territory ) Journal of Trace Elements in M edicine and Biology 21 S1: 56–58 29-M ohamedshah, F.Y.; M oser-Veillon, P.B.; Yamin, I.S.; Douglass, L.W.;Anderson, R.A. and Veillon C. (1998): Dist ribution of a stable isot op of chromium (53Cr) in serum, urine, and breast milk in lactating woman. American Journal of Clinical Nut rition, 67: 1250–1255. 30-Lifschitz , M .L.; Wallach, S. and Peabody , R.A. (1980) Radiochromium distribution in thy roid and p arathy roid deficiency. American Journal of Clinical Nut rition, 33:57–62. 32-Lee, C.R.; Yoo, C.I.; Lee, J.H. and Kang, S.K. (2002) Nasal sep tum perforation of welders. Industrial Health, 40: 286– 289. 31- Goncharov, A.T and Ametov, A.S(1977) thy roid gland morphological and functional indices in the exp erimental action of chromium .p robl E ndokrinol ,23(6):59-61 33- Valenta, L.J: (1976) Thy roid peroxidase, thy roglobulin, CAM P and DNA in human thy roid. J Clin Endocrinol M etab 43:466469, 34-M asahiro, S, ; Roy, L. ; Harri, L. and Ann, M .(1984) thy roid T4–deiodinase activit in normal and abnormal human thy roid gland .M etabolism,33(4)332-336 35-Rapop ort, B. and M cLachlan, S. A(2001).T hy roid autoimmunity .J.Clin.Invest .108:1253- 1259. 36-M arcocci, C. and Chiovato, L.( 2000) Thy roid–directed antibodies. In: Braverman LE, Ut iger RD. The Thy roid: A fundamental and clinical text. 8th ed. Philadelphia: Williams & Wilkins, p . 414-31. 37. Nagataki, S.; Uchimura, H. and M atsuy ama, Y. (1973) et al: Thy rotrop in and thy roidal p eroxidase activity . Endocrinology 92:363-371 38-. Yamamoto, K. and DeGroot , L.J. (1974) Peroxidase and NADPHcytochrome c reductase activity during thy roid hyp erplasia and involution. Endocrinology 95:606-612, 39-. Nataf, B.M .; Fragu, P. and Ot hman, S.B.(1978) Relationship between p eroxidase activity and serum TSH. T 4 and T, levels in rats in the course of iodine deficiency. 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Table (1): The mean ± S E of se rum levels of chromium and Thyroid peroxidase(TPO) activity In hyperthyroidism patients and normal subjects P-value normal subjects hyp erthy roidism p atients Parameters 0.001 51.0 ±1.8 152.0± 4.6 Thy roid peroxidase (TPO) IU/L 0.003 0.35±0.0241 2.159±0.15 Chromium pp b Table(2): The mean ± S E of se rum levels of chromium and Thyroid peroxidase(TPO) activity In hypothyroidism patie nts, and normal subjects P-value normal subjects Hyp othy roidism p atients Parameters 0.006 51.0 ±1.8 35.0± 0.31 Thy roid peroxidase (TPO) IU/L 0.001 0.35±0.0241 0.09±0.011 Chromium pp b Y=0.0216X – 1.2734 r =0.12 IBN AL- HAITHAM J. FOR PURE & APPL. S CI. VOL. 24 (2) 2011 Y=0.0028X – 0.0027 r=0.11 2011) 2( 24المجلد مجلة ابن الھیثم للعلوم الصرفة والتطبیقیة الكروم الثالثي وفعالیة انزیم بیروكسیدیز دراسة عن العالقة ما بین تركیز ایون الدرقي في امصال المصابین باختالل الوظیفة الدرقیة ور جلیل محمود ، بروین عبد الصمد اسماعیلفحامد غفوري حسن ، طی جامعة بغداد،ابن الهیثم -كلیة التربیة قسم الكیمیاء ، جامعة هاولیر الطبیة،كلیة الطب الدین جامعة صالح،كلیة التربیة 2010تشرین األول 27:استلم البحث في 2011شباط 8: قبل البحث في الخالصة تتلخص اهداف هذه الدراسة في تقدیر تركیز ایون العنصر الضئیل الكروم الثالثي وكذلك تقدیر فعالیة انزیم الوظیفة الدرقي، ومن ثم ایجاد عالقة تربط ما بین البیروكسیدیز الدرقي في مصول االشخاص المصابین بمرض اختالل ق مطیافیة االمتصاص الذري في تقدیر تركیز الكروم في عینات المصول المرضیة والطبیعیة كما ائت طر لمعاست. االثنین ELIت تقنیة ال لمعواست SA ا اوجدت النتائج ان هناك زیادة معنویة. في تقدیر فعالیة انزیم البیروكسیدیز الدرقي وارتفاع pp 0.15±2.159(ملحوظین في مستویات الكروم وفعالیة البیروكسیدیز b ) (152.0±4.6 IU/L ( على التوالي في 0.024pp±0.378(مصول المرضى المصابین بالفرط الدرقي عنه في المصل الطبیعي b ) (51.0± IU/L1.8 .( كما ة نانخفضا ا بینت النتائج ان تركیز الكروم وفعالیة البیروكسیدیز قد خفاضا ملحوظا في مصول قصور الدرقی )0.099±0.011 pp b ) (35.0±0.31 IU/L (ة . وعلى التوالي ومقارنة في امثالها عند الطبیعیین كما بینت النتائج العالق . نعند مقارنة تركیزالكروم وفعالیة البیروكسیدیز عند المرضى والطبیعیی ) P<0.001, r=0.11( المعنویة واالیجابیة