Investigation of zinc concentrations in saliva of patients with thyroid diseases: Tiroid Hastaligi Olan Kisilerde Tukuruk Cinko Konsantrasyonunun Arastirilmasi.
Purpose: It is well known that thyroid hormone disorders can influence homeostasis of trace elements in the body, such as zinc (Zn). The aim of this study was to evaluate saliva Zn levels in patients with thyroid diseases.
Material and Method: The study was performed on 126 patients (21 male, 105 female) with thyroid diseases (31 hyperthyroid, 30 hypothyroid, 31 subclinical hyperthyroid and 34 subclinical hypothyroid) and 38 healthy control subjects (7 male, 31female). Saliva was collected from all participants and Zn levels were determined on an atomic absorption spectrophotometer.
Results: Our results demonstrated that saliva Zn (p=0.007 for hypothyroid patients and p=0.023 for subclinical hypothyroid patients) levels were significantly lower in hypothyroid and subclinical hypothyroid patients than in control subjects.
Discussion: This is the first study investigating saliva Zn levels in patients with thyroid diseases. Our results show that saliva Zn levels are affected by hypothyroid disease. Turk Jem 2013; 17:1-4
Key words: Hyperthyroid; hypothyroid; subclinical hyperthyroid; subclinical hypothyroid; saliva zinc
Amac: Tiroid hormon bozukluklarinin vucutta cinko (Zn) gibi iz elementlerin homeostazina etki edebildigi cok iyi bilinmektedir. Bu calismanin amaci tiroid hastalarinda tukuruk Zn seviyelerini belirlemektir.
Gerec ve Yontem: calismamiz 126 (21E, 105K) tiroid hostasi (31 hipertiroid, 30 hipotiroid, 31 subklinik hipertiroid ve 34 subklinik hipotiroid) ve 38 (7E, 21K) saglikli kontrol uzerinde yapildi. Calismaya katilanlarin tukuruk ornekleri alinarak Zn seviyeleri atomik absorbsiyon spektrofotometresi ile olculdu.
Bulgular: Sonuclarimiz hipotiroidi (p=0.007) ve subklinik hipotiroidi (p=0.023) hastalarinin tukuruk Zn seviyelerinin kontrollerden anlamli dusuk oldugunu gosterdi.
Tartisma: Calismamiz tiroid hastalarinda tukuruk Zn seviyelerini arastiran ilk calismadir ve tukuruk Zn seviyelerinin hipotiroidi hastaligi durumundan etkilendigini gostermistir. Turk Jem 2013; 17: 1-4
Anahtar kelimeler: Hipotiroid, hipertiroid, subklinik hipotiroidi, subklinik hipertiroidi, tukuruk Zn
Abnormal thyroid function has important public health consequences (1). Thyroid hormones play an important role in human body metabolism (2). Subclinical thyroid disease is a common disorder, particularly in middle-aged and elderly individuals and the clinical significance of subclinical thyroid dysfunction is much debated (3), (4).
Mixed saliva consists mainly of water (98%), electrolytes, mucus, antibacterial constituents, and various enzymes (5). The function of the saliva glands is regulated by the sympathetic nervous system which influences saliva circulation (6). It is well known that the composition of saliva depends on a number of factors related to physiological, pathological, environmental factors, and the humoral agents, including thyroid hormones (6), (7). The mixed saliva composition may differ on account of varying contribution rate and quality from the individual glands, the presence or absence of a stimulatory source, the type of food ingested, the circadian cycles and the presence of infections (7). It is known that saliva could offer an excellent alternative to serum as a biological fluid analyzed for diagnostic purposes. This would be of great biomedical importance. Because it is very easy to collect, offering a cost-effective approach for screening of large populations, and could represent an alternative for patients whose blood is difficult to obtain or when compliance is a problem (8).
Zinc (Zn) is an essential trace element that is present in small amounts in all tissues and body fluids, including saliva (5), (7). Zn, an intracellular signaling molecule and an anti-insammatory agent, is instrumental in immune response and serves important functions in the body with its antioxidant properties (9). Thyroid hormones could affect the tissue-specific distribution and availability of some minerals. In some studies, alterations in Zn homeostasis have been reported in subjects with thyroid disorders (10). However, there are inconclusive data on alteration in Zn status in patients with thyroid diseases. In addition, to our knowledge, saliva Zn level which is a good indicator of body Zn status has not been investigated yet in patients with thyroid diseases. Therefore, in the present study, we have investigated saliva Zn levels and correlations between saliva Zn and serum thyroid hormones (TSH, free T3 and free T4) levels in patients with thyroid diseases.
Materials and Methods
This study was performed on 126 patients with thyroid function disorders (21 male, 105 female) aged 18-70 years (mean: 38.19 [+ or -] 12.50) and 38 control subjects 17 male, 31 female) aged 18-70 years (mean: 36.3 [+ or -] 10.2). Among the subjects who participated in this study, 3119 male, 22 female) had hyperthyroidism, 30 12 male, 28 female)--hypothyroidism, 31 15 male, 26)-- subclinical hyperthyroidism and 34 15 male, 29 female) had subclinical hypothyroidism. Patients with a free 13 of >4.71 pg/mL and free 14 of >1.9 ng/dL were considered to have hyperthyroidism, while those with a free T3 of <1.57 pg/mL and free T4 of <0.8 ng/dL were diagnosed with hyperthyroidism. The diagnosis of subclinical hypothyroidism was established in subjects with TSH of >4.2 [micro]U/mL and serum free T3 and free T4 levels within the reference ranges. Subjects with TSH of < 4.2 [micro]U/ ml and serum free T3 and free T4 levels within the reference ranges were considered to have subclinical hypothyroidism. The normal range for serum TSH, free T3 and free 14 levels are 0.27 to 4.2 ([micro]U/mL), 1.57 to 4.71 (pg/mL) and 0.8 to 1.9 (ng/dL), respectively in our laboratory.
Dietary patterns of the subjects were estimated using a food intake questionnaire. All subjects ate ordinary food. Less than 5% of patients with thyroid diseases and control subjects reported tobacco use. Exclusion criteria for the study included malignant disease, diabetes mellitus, chronic liver disease, chronic kidney disease, infectious disease, hypertension, a history of cardiovascular disease and taking Zn supplements. The study protocol was approved by the Ethics Committee of Meram Medical School, University of Selcuk, Konya, Turkey. All patients were informed of the details of the study and written informed consent was obtained from all subjects
Body height and weight were measured with participants wearing indoor clothing without shoes. Body mass index (BMI) was computed by dividing the body weight (in kilograms) by height (in meters) squared. Blood sampling was performed in the fasting state at 08.00 h. Serum glucose, TSH, free T3 and free T4 levels were measured immediately.
The study of saliva secretion was performed without any stimulus in the morning (09:00h-11:00h) under standard temperature and humidity conditions. All subjects refrained from eating, drinking and smoking for a minimum of 2 h before saliva collection. Subjects were comfortably seated and after a few minutes of relaxation, they were trained to avoid swallowing saliva; later they were asked to lean forward and spit all the saliva they produced for 10 min into a graduated test tube through a glass funnel. Immediately after the collection, the saliva sample was transferred to clean and minerals free microlubes and than centrifuged at 12 000 rpm for 5 min to remove the precipitate. The supernatant was frozen at -80 oC until assay.
Measurement of Zn concentrations
Zn concentrations were determined using atomic absorption spectrophotometer (Rayleigh WFX-320). For the determination of Zn in saliva, the slit width was 0.4 nm, lamp flow was 3.0 mA, wavelength was 213.9 nm. The linear range of Zn standard curve was 0.0-0.5 pg/ml. Detection limit of Zn was 0.007 pg/ml. The test repeatability was assured by rejecting the results if the percentage of Related Deviation Standard (RDS%) was more than 10%. All of the reagents were of analytical grade and water was double distilled. Just before analysis, the samples were allowed to defrost at room temperature and vigorously shaken in a vortex for 30s to rehomogenize it just before analysis. Aliquot (1mL) of saliva samples was diluted with 2 mL of 1% nitric acid (Merck) immediately before the assay. The levels of saliva Zn were calculated after application of absorbencies on suitable calibration curve for each element made from standard solutions. Concentrations of Zn elements were calculated as [micro]g/L mixed saliva volume.
Measurement of other analyses
Serum glucose was measured by commercially available kits based on routine methods on the Synchron LX System (Beckman Coulter, Fullerton CA). Serum TSH, free T3 and free T4 levels were measured by routine chemiluminesans method on E170 analyzer (Roche Diagnostics).
All data are expressed as mean [+ or -] standard deviations (SDI. Statistical analyses were done using SPSS version 16.0 for Windows. The differences between two means were analyzed by one-way analysis of variance (ANOVA). Differences were considered significant at a probability level of p<0.05.
Clinical and analytical characteristics and saliva Zn levels in patients with thyroid disorder and control subjects are presented in Table 1. As seen, serum free T3 and free T4 levels were significantly higher in hyperthyroid subjects than in controls. BMI and serum TSH levels were significantly higher and serum free T4 levels were significantly lower in hypothyroid subjects than in controls. On the other hand, serum free T3 and free T4 levels were significantly lower and serum TSH levels were significantly higher in hypothyroid subjects than in hyperthyroid ones.
Table 1. Saliva Zn levels and characteristics of thyroid disorder subjects and control subjects (1) Control Hyperthyroid Hypothyroid N 38 (7M, 31 (9M, 22F) 30 (2M, 28F) 31F) Age 34.3 [+ 38.6 [+ or -] 37.3 [+ or -] or -] 14.0 12.4 10.9 Duration of - 43 62 [+ or -] 51.28 [+ or disease (month) 34.4 -] 46.8 BMI 23.5 [+ 26 1 [+ or -] 29.3 [+ or -] [kg/[m.sup.2]) or -] 4.6 5.6 (b) 3.9 Fasting blood 87.6 [+ 92.0 [+ or -] 87.3 [+ or -] glucose (mg/dL) or -] 69.4 8.1 4.1 Free [T.sub.3] 2.87[+ or 8.89[+ or -] 2.60 [+ or -] (pg/mL) -] 0.5 4.7 (b) 0.9 (d) Free[T.sub.4] 1.20 [+ 3.48 [+ or -] 0.60 [+ or -] ng/dL) or -] 1.6 (b) 0.1 (a**), 0.1 (d) TSH 2.20 [+ 0.01 [+ or -] 54.76 [+ or [[micro]U/mL] or -] 0.01 -] 23.6 (b), 1.0 (d) Saliva zinc (Zn) 285.1 [+ 258.6 [+ or -] 163 2 [+ or ([micro]g/L) or -] 182.9 -] 68.80 220.1 (a*) Subclinical Subclinical Mean hyperthyroid hypothyroid comparison (P) N 31 (5M, 26F) 34 (5M, 29F) - Age 40.6 [+ or -] 36.5 [+ or -] 0.192 11.7 11.8 Duration of 3.81 [+ or -] 4,62 [+ or -] - disease (month) 2.41 2.64 BMI 26.3 [+ or -] 28.1 [+ or -] p < 0.001 [kg/[m.sup.2]) 6.3 5.6 (a*) Fasting blood 92.4 [+ or -] 91.1 [+ or -] 0.057 glucose (mg/dL) 8.8 8.8 Free [T.sub.3] 3.75 [+ or -] 3.41 [+ or -] p < 0.001 (pg/mL) 0.5 (d) 0.5 (d) Free[T.sub.4] L39 [+ or -] 1.15 [+ or -] p < 0.001 ng/dL) 0.2 (d), (e) 0.2 (d), (g) TSH 0.12 [+ or -] 9.58 [+ or -] p < 0.001 [[micro]U/mL] 0.08 (f), (m) 4.2 (a**), (d), (f) Saliva zinc (Zn) 260.9 [+ or -] 1719 [+ or -] 0.003 ([micro]g/L) 161.2 82.5 (c) (1) All values (except gender) are mean [+ or -] standart deviations. BMI, body mass index; [T.sub.3], triodothyronine; [T.sub.4], thyroxine; TSH. thyroid-stimulating hormone; a*p = 0.007, a**p = 0.017, bp< 0.001, cp = 0.023, compared with control group; dp < 0.001 compared with hyperthyroid group; ep= 0.00, fp < 0.001, gp = 0.039 compared with hypothyroid group, mp = 0.004 compared with subclinical hypothyroid group
Furthermore, BMI and serum TSH levels were significantly higher in subclinical hypothyroid subjects than in controls. Saliva Zn levels were significantly lower in hypothyroid and subclinical hypothyroid subjects than in control subjects. On the other hand, saliva Zn levels did not correlate with serum thyroid hormone levels (TSH, free T3 and free T4) in the groups (Table 2).
Table 2. The correlations of serum thyroid hormon levels with saliva Zn levels of the groups Control Hyperthyroid Hypothyroid Subclinical hyperthyroid Saliva Zinc Free r = r = -0.143 r = 0.119 r = -0.121 [T.sub.3] 0.105 p = p = 0.487 p = 0.563 p = 0.495 0.580 Free r = r = -0.189 r = -0.365 r = -0.230 [T.sub.4] 0.308 p = p = 0.355 p = 0.067 p = 0.192 0.097 TSH r = r = 0.173 r = -0.118 r = -0.239 0.033 p = p = 0.397 p = 0.567 p = 0.173 0.861 Subclinical hypothyroid Free r = 0.069 [T.sub.3] p = 0 696 Free r = 0.112 [T.sub.4] p = 0.527 TSH r = 0.208 p = 0.237 Free [T.sub.3], triodothyronine; free [T.sub.4], thyroxine: TSH, thyroid-stimulating hormone
In this study, we found that hypothyroid and subclinical hypothyroid group had a significantly decreased saliva Zn levels compared with the control group. Besides, to our knowledge, saliva Zn levels in patients with thyroid diseases have not been investigated yet. As serum levels of a variety of electrolytes are correlated to saliva levels of these electrolytes, it is possible that some of the factors that affect serum Zn levels will also affect saliva Zn levels (11).
There are a few works in the literature investigating the level of serum Zn in patients with thyroid diseases. Although there are some conflicting findings, generally, the findings of the other investigators support ours. For example, Al-juboori et al. (2) Zhang et al. (12), and Dolev et al. (13) have found that serum Zn levels were lower in hypothyroidism patients than in control subjects. In addition, Buchinger et al. (14) have found that hypothyroid patients had a most significantly reduced Zn content in serum compared to euthyroid patients. However, Aihara et al. (15) have found that there was no difference in plasma Zn concentrations between patients with thyroid diseases and control subjects.
It has been reported that various factors including diet, stress, infection, age, pregnancy, medications and oral contraceptives affect serum Zn levels (11). Possible explanations for these findings were reported to be severely impaired gastrointestinal absorption of Zn and a change in Zn distribution in subjects with hypothyroidism (2). It is known that, a low serum Zn level due to change in Zn distribution may reflect sequestration of Zn by the liver or other tissues. Besides, this status may be due to significant influence of TSH on the variation of the concentration of iodine, selenium and Zn in normal and altered human thyroid tissue (2). On the other hand, changes in body metabolic rate have been shown to be reflected in altered Zn metabolism. For example, thyroxine administration to rats was reported to significantly enhance the rate of Zn-turn-over and altered its distribution in various organs leading to an almost 50% depletion in soft tissue levels of Zn. Zn turnover and utilization were significantly arrested in rats made hypothyroid by iodine treatment (15). We concluded that decreased saliva Zn levels are the cause of hypothyroid disease. There are some limitations in our study. One of these limitations is the small sample size. Besides, parallel measurement of Zn both in plasma and saliva may provide more informative results.
In conclusion, our results show that saliva Zn levels are affected by hypothyroidism. Also, further studies involving larger number of subjects and parallel measurement of both in plasma and saliva may provide additional information regarding the treatment of thyroid diseases.
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Sevil Kurban, Idris Mehmetoglu, Fatma Humeyra Yerlikaya, Salt Gonen *
University of N. Erbakan, Meram Faculty of Medicine. Department of Biochemistry, Konya, Turkey
* University of N. Erbakan, Meram Faculty of Medicine, Department of Internal Medicine, Division of Endocrinology and Metabolism, Konya, Turkey
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|Title Annotation:||Original Article Ozgun Arastirma|
|Author:||Kurban, Sevil; Mehmetoglu, Idris; Yerlikaya, Fatma Humeyra; Gonen, Sait|
|Publication:||Turkish Journal of Endocrinology and Metabolism|
|Date:||Mar 1, 2013|
|Next Article:||Role of vitamin D in the immune system: D Vitamininin immun Sistemdeki Rolu.|