Comparison of potential protective effects of melatonin and propylthiouracil against lipid peroxidation caused by nitrobenzene in the thyroid gland.

BACKGROUND: Nitrobenzene is a carcinogen, which induces-among others-thyroid tumors. Melatonin is an effective antioxidant, whereas some antioxidative effects of propylthiouracil (PTU; an antithyroid medication used for the treatment of thyrotoxicosis) were also found. The aim of the study was to compare protective effects of melatonin and PTU against lipid peroxidation in homogenates of porcine thyroids, incubated in the presence of nitrobenzene. METHODS: Homogenates of porcine thyroids were incubated for 30 min in the presence of nitrobenzene (0.001, 0.01, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 7.5, and 10.0 mM). The level of lipid peroxidation products (malondialdehyde + 4-hydroxyalkenals) was measured spectrophotometrically. Nitrobenzene (7.5 and 10.0 mM) increased lipid peroxidation in the homogenates of porcine thyroids. Subsequently, homogenates of porcine thyroids were incubated for 30 min in the presence of nitrobenzene (7.5 mM) plus one of the antioxidants: melatonin (0.000001, 0.00001, 0.0001, 0.001, 0.01, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, and 7.5 mM) or PTU (0.01, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, and 7.5 mM). RESULTS: Lipid peroxidation caused by nitrobenzene was effectively prevented by melatonin, with the lowest effective concentration of 0.0001 mM, being only two orders of magnitude higher than physiological blood concentration in humans. At the same time, PTU revealed protective effects only in the highest used concentration (7.5 mM), which is practically never reached during pharmacological treatment in patients with thyrotoxicosis. CONCLUSIONS: Melatonin can serve as an effective agent in protection against nitrobenzene-induced lipid peroxidation in porcine thyroid.

Direct contribution of obesity to oxidative damage to macromolecules.

BACKGROUND: Obesity constitutes a common modifiable risk factor for certain non-communicable diseases (NCDs) associated with enhanced oxidative stress. OBJECTIVES AND METHODS: The aim of the study was to examine serum concentrations of malondialdehyde + 4-hydroxyalkenals (MDA+4-HDA), as an index of lipid peroxidation (LPO), and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) concentration in peripheral blood lymphocytes, as an index of nuclear DNA damage, in overweight and obese adult patients. LPO and 8-oxodG, as well as clinical and laboratory parameters, which are frequently affected by obesity, were evaluated in 58 overweight and obese adult patients, and in 20 healthy volunteers. RESULTS: Both LPO and 8-oxodG levels were increased in overweight and obese patients, with further increase observed with the increasing body mass index (BMI). LPO correlated positively with body mass, BMI, waist circumference, hip circumference, waist:hip ratio, systolic or diastolic blood pressure, glucose, C-reactive protein and ferritin concentrations. 8-oxodG correlated positively with body mass, BMI, hip circumference and triglyceride concentration, whereas it correlated negatively with iron concentration. Expectedly, positive correlation between LPO and 8-oxodG was also found. CONCLUSIONS: BMI constituted the only independent determinant (predictor) of LPO in overweight and obese patients. Consistently, LPO did constitute the only independent determinant of obesity. Overweight and obesity in adults are directly associated with increased oxidative damage to macromolecules.

Relationship between lipid peroxidation or carcinoembryonic antigen and risk factors for non-communicable diseases in women at midlife and beyond.

BACKGROUND: Non-communicable diseases (NCDs) constitute leading cause of morbidity, disability and premature mortality. Oxidative processes are involved in the pathogenesis of NCDs. OBJECTIVES: To investigate the relationship between lipid peroxidation (LPO), an index of oxidative damage to membrane lipids, or carcinoembryonic antigen (CEA), a tumor marker, and potential risk factors for NCDs in women at midlife and beyond. METHODS: Data on lifestyle, such as dietary habits, smoking, physical activity, etc. and medical history, were assessed by a questionnaire in 323 female outpatients of the Regional Centre of Menopause and Osteoporosis - Outpatient Department of Endocrinology, Lodz (Poland), at midlife and beyond. Blood serum LPO and CEA levels, as well as anthropometric measurements were evaluated. RESULTS: Positive correlations between LPO level and body mass or body mass index or hip circumference were found. LPO level was increased in women who did not declare regular menstrual cycles. CEA level was increased in women who smoked (and positively correlated with duration of smoking), who consumed pickled food every day and over-consumed animal fats, who had not breastfed in the past, as well as in women with malignancy in anamnesis. Logistic regression analysis has revealed that LPO constitutes the independent positive determinant, whereas CEA constitutes the independent negative determinant, of obesity. Moreover, CEA was independently associated with malignancy in anamnesis, cigarette smoking and animal fat over-consumption. CONCLUSION: Both LPO and CEA are independently associated with certain modifiable risk factors for NCDs.

Melatonin restores the basal level of lipid peroxidation in rat tissues exposed to potassium bromate in vitro.

OBJECTIVE: Potassium bromate (KBrO3) is a prooxidant and carcinogen. Melatonin is a highly effective antioxidant. Indole-3-propionic acid (IPA; indole substance) and propylothiouracil (PTU; antithyroid drug) reveal some antioxidative effects. The aim of the study was to evaluate KBrO3-induced lipid peroxidation (LPO) in vitro in tissues collected from control or melatonin-treated rats, and to compare potential preventive effects of melatonin, IPA and PTU. MATERIALS AND METHODS: Kidney, liver and lung homogenates from either control or melatonin-pretreated rats (0.0645 mmol/kg b.w., i.p., twice daily, 10 days) were incubated in the presence of KBrO3 (0.1, 0.5, 1.0, 2.5, 5.0, 10.0 mM). Then, control lung homogenates were incubated with KBrO3 (10.0 mM) together with melatonin (0.01, 0.1, 0.5, 1.0, 5.0, 7.5 mM), or with IPA or with PTU. LPO products (malondialdehyde+4-hydroxyalkenals) were measured spectrophotometrically. RESULTS: Melatonin injections prevented KBrO3-induced LPO in lung homogenates. Melatonin, IPA and PTU, used in vitro, reduced KBrO3-induced LPO in control lungs. Unexpectedly, KBrO3 caused a concentration-dependent decrease in LPO in liver and kidney homogenates from control but not from melatonin-treated rats. CONCLUSIONS: Potassium bromate-induced LPO in the rat lung homogenates suggests that the lung may be the target for this carcinogen. An exposure of organisms to melatonin decreases tissue sensitivity to KBrO3-induced damage, possibly by restoring the oxidative balance.

External hydrogen peroxide is not indispensable for experimental induction of lipid peroxidation via Fenton reaction in porcine ovary homogenates.

OBJECTIVE: Substrates of Fenton reaction (Fe(2+)+H(2)O(2)-->Fe(3+)+*OH+OH-) may be used to experimentally induce oxidative damage to macromolecules. The study aimed at evaluating effects of Fe(2+) and/or H(2)O(2) on lipid peroxidation in porcine ovary homogenates. MATERIALS AND METHODS: Ovary homogenates were incubated in the presence of either H2O2 (100, 50, 25, 10, 5.0, 2.5, 1.0, 0.5, 0.25, 0.01, 0.001 mM) or FeSO(4) (Fe2+) (300, 150, 75, 30, 15, 7.5, 3.0, 1.5, 0.75 microM), or of those two factors used together: Fe(2+) (30 microM) plus H(2)O(2) (concentrations as above), or H(2)O(2) (0.5 mM) plus Fe(2+) (concentrations as above). The concentration of malondialdehyde+4-hydroxyalkenals constituted the lipid peroxidation index. RESULTS: H(2)O(2) alone did not affect lipid peroxidation in porcine ovary homogenates at all, whereas Fe(2+) (300, 150, 75, 30, and 15 microM) alone increased lipid peroxidation in a concentration dependent manner. When Fe(2+) and H(2)O(2) were applied together, lipid peroxidation increased significantly without any concentration related effect of H(2)O(2), but with a clear concentration dependent effect of Fe(2+); the damaging effect of Fe(2+), used together with H(2)O(2), was the same as the one, obtained after Fe(2+) was applied alone. CONCLUSIONS: In conclusion, external H(2)O(2) is not indispensable for experimental induction of lipid peroxidation by Fenton reaction in porcine ovary homogenates.

Pyrimidine and purine salvage deoxyribonucleoside metabolism in hepatic and renal homogenates from rats pretreated with propylthiouracil or L-thyroxine.

OBJECTIVES: In vitro activities of thymidine kinase (TK, EC 2.7.1.21), adenosine kinase (AK, EC 2.7.1.20) and deoxycytidine kinase (dCK, EC 2.7.1.74) enzymes involved in the salvage pathway of DNA precursor synthesis, in homogenates of the rat liver and kidney, were examined. Type I iodothyronine-5'-deiodinase (5'D-I) is the main enzyme responsible for peripheral metabolism of thyroid hormones. This occurs especially in the liver, kidney and muscle. The activity of 5'D-I is inhibited bypropylthiouracil (PTU), an antithyroid drug. METHODS: The liver and kidney were collected from rats pretreated in vivo with either a 0.1% solution of PTU in drinking water for 2 weeks or injected with levothyroxine (L-T(4), 50 &mgr;g/kg BW, daily) for 2 weeks. The enzyme activities were measured by ascending chromatography and expressed asthe amounts of radioactive reaction products of the phosphorylation of dThd (for TK), ofdAdo (for AK and dCK) and of dGuo (for dCK). RESULTS: In liver homogenates, PTU-pretreatment decreased the activities of the three enzymes when compared to control values and those of L-T(4)-treated animals; also L-T(4) injections decreased the AK and dCK activities in the liver homogenates. PTU-pretreatment increased TK activity and the rate of dGuo phosphorylation in kidney homogenates, when compared to controls and to the L-T(4)-pretreated animals. Conversely, both PTU- and L-T(4)-pretreatment reduced the rate of dAdo phosphorylation in kidney homogenates. CONCLUSION: Changes in the activities of examined enzymes which participate inpyrimidine orpurine metabolism of the salvage pathway of DNA synthesis in the liver afterPTU-pretreatment (as shown herein) are similar to the changes of the 5'D-I activity after PTU-treatment (as reported by others). Thus, the observations suggest a role of the salvage pathway of DNA synthesis in the peripheral metabolism of thyroid hormones.

Protective effects of melatonin and indole-3-propionic acid against lipid peroxidation, caused by potassium bromate in the rat kidney.

Potassium bromate (KBrO(3)) is classified as a carcinogenic agent. KBrO(3) induces tumors and pro-oxidative effects in kidneys. Melatonin is a well known antioxidant and free radical scavenger. Indole-3-propionic acid (IPA), an indole substance, also reveals antioxidative properties. Recently, some antioxidative effects of propylthiouracil (PTU)-an antithyroid drug-have been found. The aim of the study was to compare protective effects of melatonin, IPA, and PTU against lipid peroxidation in the kidneys and blood serum and, additionally, in the livers and the lungs, collected from rats, pretreated with KBrO(3). Male Wistar rats were administered KBrO(3) (110 mg/kg b.w., i.p., on the 10th day of the experiment) and/or melatonin, or IPA (0.0645 mmol/kg b.w., i.p., twice daily, for 10 days), or PTU (0.025% solution in drinking water, for 10 days). The level of lipid peroxidation products-malondialdehyde + 4-hydroxyalkenals (MDA + 4-HDA)-was measured spectrophotometrically in thyroid homogenates. KBrO(3), when injected to rats, significantly increased lipid peroxidation in the kidney homogenates and blood serum, but not in the liver and the lung homogenates. Co-treatment with either melatonin or with IPA, but not with PTU, decreased KBrO(3)-induced oxidative damage to lipids in the rat kidneys and serum. In conclusion, melatonin and IPA, which prevent KBrO(3)-induced lipid peroxidation in rat kidneys, may be of great value as protective agents under conditions of exposure to KBrO(3).

Comparison of potential protective effects of melatonin, indole-3-propionic acid, and propylthiouracil against lipid peroxidation caused by potassium bromate in the thyroid gland.

Potassium bromate (KBrO3) is a prooxidant and carcinogen, inducing thyroid tumors. Melatonin and indole-3-propionic acid (IPA) are effective antioxidants. Some antioxidative effects of propylthiouracil (PTU)--a thyrostatic drug--have been found. The aim of the study was to compare protective effects of melatonin, IPA, and PTU against lipid peroxidation in the thyroids, collected from rats treated with KBrO3, and in homogenates of porcine thyroids, incubated in the presence of KBrO3. Wistar rats were administered KBrO3 (110 mg/kg b.w., i.p., on the 10th day of the experiment) and/or melatonin, or IPA (0.0645 mmol/kg b.w., i.p., twice daily, for 10 days), or PTU (0.025% solution in drinking water, for 10 days). Homogenates of porcine thyroids were incubated for 30 min in the presence of KBrO3 (5 mM) plus one of the antioxidants: melatonin (0.01, 0.1, 0.5, 1.0, 5.0, 7.5 mM), or IPA (0.01, 0.1, 0.5, 1.0, 5.0, 7.5, 10.0 mM), or PTU (0.01, 0.1, 0.5, 1.0, 5.0, 7.5, 10.0 mM). The level of lipid peroxidation products (MDA + 4-HDA) was measured spectrophotometrically in thyroid homogenates. In vivo pretreatment with either melatonin or with IPA or with PTU decreased lipid peroxidation caused by KBrO3--injections in rat thyroid gland. Under in vitro conditions, PTU (5.0, 7.5, and 10.0 mM), but neither melatonin nor IPA, reduced KBrO3-related lipid peroxidation in the homogenates of porcine thyroids. In conclusion, melatonin and IPA may be of great value as protective agents under conditions of exposure to KBrO3.

Expression of genes for certain enzymes of pyrimidine and purine salvage pathway in peripheral blood leukocytes collected from patients with Graves' or Hashimoto's disease.

Increased activities of some enzymes, which participate in pyrimidine and purine salvage pathway, were found in blood fractions of patients suffering from different autoimmunological diseases, thyroid diseases included. The aim of the study was to estimate the expression of genes, specific for deoxycytidine kinase (dCK, EC 3.7.1.74), thymidine kinase 1 (TK1; EC 2.7.1.21), and adenosine deaminase (ADA, EC 3.5.4.4) in blood leukocytes, collected from patients with autoimmunological thyroid diseases (AITD), i.e., Graves' or Hashimoto's disease. The total mRNA was isolated from peripheral blood leukocytes and, afterwards, submitted to reverse transcription (RT), with the following amplification of genes encoding for particular examined enzymes and beta-actin, as a supervisory gene [RT-polymerase chain reaction (RT-PCR)]; ADA gene was amplified with the use of three different primer pairs (ADA3, ADA4, and ADA5). PCR products were electrophoresed in 8% polyacrylamide gel and then, submitted to densitometric analysis. The levels of expression of all the examined genes in leukocytes from patients with either Graves' or Hashimoto's disease were significantly increased when compared to those in controls; above a twofold elevation of expression of TK1, ADA4, and ADA5 genes was observed. In conclusion, the changes of activities of salvage enzymes in patients with AITD occur likely at transcription level; the measurement of gene expression for purine and pyrimidyne salvage enzymes may likely help explain the mechanism of autoimmune diseases, being also significant in the diagnostics and/or monitoring of AITD.

Purine metabolism in leukocytes and erythrocytes in Graves' or Hashimoto's disease.

INTRODUCTION: Adenosine deaminase (ADA), purine nucleoside phosphorylase (PNPase), S-adenosylhomocysteine hydrolase (SAHH), 5'-nucleotidase (5N), and deoxycytidine kinase (dCK) are involved in purine salvage metabolism. Changes of the activities of the above enzymes have been observed in blood cells in patients with immunological disorders. MATERIALS AND METHODS: The activities of ADA, PNPase, SAHH, 5'N, and dCK in lysates of leukocytes and erythrocytes, obtained from patients with Graves' or Hashimoto's disease, were measured, using chromatographic analysis. Serum concentrations of antithyroglobulin (Tg Ab) and antithyroperoxidase (TPO Ab) antibodies were measured by an immunoenzymatic method. RESULTS: (1) ADA activity in leukocytes, obtained from patients with Hashimoto's disease, was significantly higher than in control leukocytes, as well as in leukocytes from patients with Graves' disease; (2) dCK activities in leukocytes from patients with both Graves' and Hashimoto's diseases were approximately four and five times higher, respectively, than in leukocytes of control subjects; (3) a positive correlation was observed between dCK activity in leukocytes and serum Tg Ab concentration in patients with Graves' disease. In conclusion, the increased ADA and dCK activities in leukocytes from patients with Graves' and Hashimoto's diseases may be regarded as indicators of autoimmunological thyroid diseases.