The effect of Apigenin on glycometabolism and cell death in an anaplastic thyroid cancer cell line.

AIMS AND BACKGROUND: A more pronounced characteristic of cancer cells is the energy dependence on glucose, which mitigated by glucose transporters. The comprehension of the regulatory mechanisms behind the Warburg effect holds promise for developing therapeutic interventions for cancers. Studies are lacking which targeted the GLUTs for treatment of malignancy of thyroid tumors. In our current investigation, we have undertaken this study to determine the potential of Apigenin, plant derived flavonoid in modulating tumor apoptosis by targeting GLUTs expression in SW1736 cell line of anaplastic thyroid carcinoma. MATERIAL METHODS: Flow cytometry with propidium iodide staining was used to determine cell apoptosis. For glucose uptake detection, the "GOD-PAP" enzymatic colorimetric test was used to measure the direct glucose levels inside the cells. To determine the expression of GLUT1 and GLUT3 mRNA in the SW1736 cell line qRT-PCR was employed. Protein levels of GLUT1 and GLUT3 in the SW1736 cell line were detected with western blotting. Also, the scratch wound healing assay was conducted for cell migration. RESULTS: According to qRT-PCR analysis, the levels of GLUT1 and GLUT3 mRNA were lower in the group that received Apigenin relative to the control group. The Apigenin treatment of SW1736 cells decreased protein expression of the GLUT1 and GLUT3 levels in conformity to qRT-PCR. The scratch assays revealed that Apigenin treatment of cancer cell lines inhibited cell migration as compared to control. CONCLUSION: These findings demonstrate the possibility of targeting the glucose facilitators' pathway for making thyroid cancer cells more susceptible to programmed cell death.

The effect of homocysteine thiolactone on paraoxonase and aryl esterase activity of human serum purified paraoxonase 1 in vitro experiments.

BACKGROUND: The important role of lipoproteins, particularly low-density lipoprotein (LDL) and high-density lipoprotein (HDL), has been highly regarded among the known causes of cardiovascular disease (CVD). A wide range of risk factors may cause structural and functional changes in lipoprotein particles, resulting in deposition and formation of atherosclerotic plaques. Homocysteine is one of the most important risk factors in heart disease, and its atherosclerotic properties appear to be related to its intermediate metabolite called homocysteine thiolactone (HCTL). The major aim of the present investigation was to study the effect of HCTL in different concentrations (10, 50, and 100 µM) on paraoxonase and aryl esterase activities of purified human serum paraoxonase 1 (PON1) antioxidant enzyme related to HDL, as an extracellular hydrolyzing enzyme of HCTL. METHODS: In order to purify PON1 enzyme from human serum, three-step chromatographic methods including DEAE Sephadex A50, Sephadex G100, and DEAE Sephadex A50 were used. Protein concentration and paraoxonase and aryl esterase activities of each fraction were measured separately and the highest activities fractions were collected and subsequently pooled together for the next steps. Ultimately, both activities of PON1 in the presence of different concentrations of HCTL were measured in triplicate by spectrophotometry technique. RESULTS: HCTL at concentrations of 50 and 100 µM decreased both paraoxonase and aryl esterase activities (P < 0.05) in comparison with the control group, which is directly related to the increase in HCTL concentration. However, at a concentration of 10 µM HCTL, no significant difference was observed in both paraoxonase and aryl esterase activities compared to the control group. CONCLUSION: HCTL is a highly toxic and reactive compound that is produced in all cells. Extracellular enzyme PON1 causes its hydrolysis with high efficiency. The results obtained from the present study showed that paraoxonase and aryl esterase activities decreased in vitro in the presence of HCTL and therefore, HCTL may cause changing in the protein structure of this enzyme. Previous in vivo studies have also shown decrease of PON1 activity in patients with hyperhomocysteinemia.

Regulators of glucose uptake in thyroid cancer cell lines.

Thyroid cancer is the most common sort of endocrine-related cancer with more prevalent in women and elderly individuals which has quickly widespread expansion in worldwide over the recent decades. Common features of malignant thyroid cells are to have accelerated metabolism and increased glucose uptake to optimize their energy supply which provides a fundamental advantage for growth. In tumor cells the retaining of required energy charge for cell survival is imperative, indeed glucose transporters are enable of promoting of this task. According to this relation it has been reported the upregulation of glucose transporters in various types of cancers. Human studies indicated that poor survival can be occurred following the high levels of GLUT1 expression in tumors. GLUT-1 and GLUT3 are the glucose transporters which seems to be mainly engaged with the oncogenesis of thyroid cancer and their expression in malignant tissues is much more than in the normal one. They are promising targets for the advancement of anticancer strategies. The lack of oncosuppressors have dominant effect on the membrane expression of GLUT1 and glucose uptake. Overexpression of hypoxia inducible factors have been additionally connected with distant metastasis in thyroid cancers which mediates transcriptional regulation of glycolytic genes including GLUT1 and GLUT3. Though the physiological role of the thyroid gland is well illustrated, but the metabolic regulations in thyroid cancer remain evasive. In this study we discuss proliferation pathways of the key regulators and signaling molecules such as PI3K-Akt, HIF-1, MicroRNA, PTEN, AMPK, BRAF, c-Myc, TSH, Iodide and p53 which includes in the regulation of GLUTs in thyroid cancer cells. Incidence of deregulations in cellular energetics and metabolism are the most serious signs of cancers. In conclusion, understanding the mechanisms of glucose transportation in normal and pathologic thyroid tissues is critically important and could provide significant insights in science of diagnosis and treatment of thyroid disease. Video Abstract.

Cadmium Testicular Toxicity in Male Wistar Rats: Protective Roles of Zinc and Magnesium.

Cadmium (Cd) is a highly toxic element, which may cause toxicity to most organs in the body. Zinc (Zn) and magnesium (Mg) are essential minerals with probable benefits on Cd harmful effects. Finding an efficient and non-pathological treatment against Cd toxicity seems promising. Fifty adult rats were divided into ten experimental groups of five rats each. The Cd group was treated with 1 mg Cd/kg and the control group received 0.5 cm3 normal saline. The other eight groups received Zn (0.5 and 1.5 mg/kg) and Mg (0.5 and 1.5 mg/kg) either alone or in combination with 1 mg Cd/kg through IP injection for 3 weeks. Testis malondialdehyde (MDA), sperm parameters, and testis histopathology were investigated. Cd reduced sperm parameters and increased testis MDA. Moreover, Cd exposure caused a significant histological damage in testis of male rats. However, Zn or Mg treatment prevented and reversed Cd toxic alterations in testis. These findings suggest that co-administration of Zn or Mg could improve cadmium testicular toxicity in male Wistar rats.

Protective Role of Zinc and Magnesium against Cadmium Nephrotoxicity in Male Wistar Rats.

Cd is a toxic metal that has a destructive impact on most organ systems. This work aims to determine Zn or Mg protective effects against Cd renal toxicity. In this study, rats were divided into six groups. The Cd group was treated with 1 mg Cd/kg, and the control group received 0.5 cm3 normal saline, intraperitoneally. The other four groups received one of the following dosages of 1 mg/kg Cd + 0.5 mg/kg Zn, 1 mg/kg Cd + 1.5 mg/kg Zn, 1 mg/kg Cd + 0.5 mg/kg Mg, or 1 mg/kg Cd + 1.5 mg/kg Mg through IP injection for 3 weeks. Kidney malondialdehyde (MDA) and serum sodium, potassium, urea, creatinine, and protein were measured. Light microscopic examination was used for histological studies. Cd reduced serum creatinine and protein, and increased urea, sodium, and potassium. Moreover, Cd exposure caused a significant enhancement in MDA levels as well as histological damage in kidneys. Zn or Mg treatment prevented and reversed toxic alterations induced by Cd. These results suggest that Zn and Mg may have protective effects against Cd renal toxicity.

Catecholamine Contents of Different Region of Adult Rat Brain Are Altered Following Short and Long-term Exposures to Pb(+2.).

Catecholamine is a group of neurotransmitters that is believed to be responsible for the normal function of animal brain. Physiological and behavioral changes of human body have been reported due to the damage of the brain function following lead exposure. Due to the assumption of lead disposal in brain tissue with two year for its half-life, which results in alteration of brain function, we investigated the ability of lead to change the brain catecholamines during short and long-term studies. Rats were exposed daily with varying amounts of lead and catecholamine contents of cerebellum, mid-brain and brain cortex were determined. Acute peritoneal administration of single dose of lead as lead acetate (260 µmol/Kg) after 2 h reduced (p < 0.05) the catecholamine levels of cerebellum, mid-brain and cortex part by 34.9%, 35.44% and 23.8%, respectively. The extension of experiment time to 5 h, significant (p < 0.05) reductions in catecholamine levels of mentioned regions of brain by 32.35%, 12.35% and 19.3% were seen respectively. Daily intraperitoneal administration of 10 µmol/Kg lead for 30 and 60 days reduced catecholamines levels of cerebellum (22.22% and 30.44%), midbrain (12.48% and 26.27%) and brain cortex (11.58% and 26.7%) respectively. It might be concluded that brain dysfunction in lead intoxicated rat occurred through the reduction in the catecholamine levels of different parts of brain. Lead might be therefore considered as a probable factor in causing neurological disease in lead exposed man.

Changes in serum parameters associated with iron metabolism in male rat exposed to lead

Due to the severe hazardous influences of lead (Pb2+) on iron-related diseases, the effects of Pb2+ on serum parameters associated with iron metabolism have been studied in this project. Male Wistar rats weighing 200–250 g were treated with Pb2+ for the short and long period of times. The animals received daily intraperitoneal injection of 100 mg Pb2+ kg−1 body weight (BW) for 5 days and 4 mg kg−1 BW of Pb2+ for 30 and 45 days, respectively. The results show that when animals were treated with both low and high concentrations of Pb2+, serum iron concentration decreased markedly, by 23.2, 32.8, and 39.9 %, while the sera TIBC and transferrin concentrations increased significantly (p < 0.05). Following short- and long-term exposures to Pb2+, the percentage of serum transferrin saturation was also decreased in comparison with the untreated control group (p < 0.05). Concentrations of serum copper and ceruloplasmin following Pb2+ treatments also reduced significantly (p < 0.05). The percentage of hematocrit and hemoglobin levels was reduced (p < 0.05) in all Pb2+-treated animals in comparison with the controls. These results suggest that Pb2+ changes the serum parameters related to iron metabolism, which may play an important role in producing iron-related diseases (AU)

Protective effects of copper against aluminum toxicity on acetylcholinesterase and catecholamine contents of different regions of rat's brain.

The probable protective effects of copper on the acetylcholinesterase activity and the catecholamine levels in cerebellum, cortex and mid-brain of rat, which was intoxicated by aluminum, were studied during short and long terms. In this respect, male Wistar rats weighing 200-250 g were received daily intraperitoneal doses of aluminum, copper and also combined doses of both metals for 15 days (Al 10 mg kg(-1) BW and Cu 1 mg kg(-1) BW), 30 days (Al 5 mg kg(-1) BW and Cu 0.5 mg kg(-1) BW) and 60 days (Al 1 mg kg(-1) BW and Cu 0.1 mg kg(-1) BW), respectively. The results obtained from the short period of exposure (15 days) showed that aluminum produced significant (P < 0.05) decreases in the acetylcholinesterase activity by 24.14, 23.30 and 25.81 %. Similarly, the catecholamine levels were reduced by 10.69, 12.25 and 12.64 % in cerebellum, cortex and mid-brain, respectively. Treatment with copper increases both acetylcholinesterase activity and catecholamine contents of cerebellum, cortex and mid-brain. Simultaneous injection of copper and aluminum increased both acetylcholinesterase activity and catecholamine contents in all three parts of rat brain when compared to aluminum-treated group. Same results were also observed following 30 and 60 days of exposures. In overall, it has been found that copper may have a protective-like ability to hinder aluminum toxicity in the brain.

Changes in serum parameters associated with iron metabolism in male rat exposed to lead.

Due to the severe hazardous influences of lead (Pb(2+)) on iron-related diseases, the effects of Pb(2+) on serum parameters associated with iron metabolism have been studied in this project. Male Wistar rats weighing 200-250 g were treated with Pb(2+) for the short and long period of times. The animals received daily intraperitoneal injection of 100 mg Pb(2+) kg(-1) body weight (BW) for 5 days and 4 mg kg(-1) BW of Pb(2+) for 30 and 45 days, respectively. The results show that when animals were treated with both low and high concentrations of Pb(2+), serum iron concentration decreased markedly, by 23.2, 32.8, and 39.9 %, while the sera TIBC and transferrin concentrations increased significantly (p < 0.05). Following short- and long-term exposures to Pb(2+), the percentage of serum transferrin saturation was also decreased in comparison with the untreated control group (p < 0.05). Concentrations of serum copper and ceruloplasmin following Pb(2+) treatments also reduced significantly (p < 0.05). The percentage of hematocrit and hemoglobin levels was reduced (p < 0.05) in all Pb(2+)-treated animals in comparison with the controls. These results suggest that Pb(2+) changes the serum parameters related to iron metabolism, which may play an important role in producing iron-related diseases.

Liver aspartate transaminase isoenzymes as biomarkers of chronic exposure to chromium(VI).

Exposure to hexavalent chromium compounds is associated with the risk of lung cancer, dermatitis, gastrointestinal ulcers, and other tissue damages. The aim of this study was to compare liver isoenzyme and total serum activities of aspartate aminotransferase (AST) as cytotoxic biomarkers of acute and chronic cytotoxicity of Cr(VI). We investigated the extent of cell damage caused by chromium(VI) in acute (2.5 mg kg(-1)) daily doses administered over five days and chronic (0.25 mg kg(-1) and 0.5 mg kg(-1)) daily doses administered over 15 to 60 days by measuring total AST in serum and low molecular weight AST (LMW-AST) and high molecular weight AST (HMW-AST) activities in thirty liver fractions. We also evaluated the kinetic properties and electrophoretic mobility of the LMW- and HMW-AST isoenzymes in liver subcellular fractions. Liver LMW-AST and total serum AST activities significantly decreased after 15 days of exposure (P<0.05). With continued treatment, AST activity increased by 15.67% (P<0.05). Interestingly, changes in serum AST activity were similar to changes in the liver LMW-AST isoenzyme. Our results confirmed that total serum AST activity may serve as a reliable tissue biomarker for long-term exposures to Cr(VI), but they also suggest that the LMW-AST isoenzyme could be even more sensitive.

Protective effect of zinc on related parameters to bone metabolism in common carp fish (Cyprinus carpio L.) intoxified with cadmium.

The short term effects of waterborne cadmium (Cd(+2)) on the levels of serum parameters related to bone metabolism including calcium (Ca), inorganic phosphorus (P(i)) and alkaline phosphatase (ALP) in common carp fish (Cyprinus carpio L.) were studied. Fish were treated with varying concentrations of Cd(+2) (0.22, 1.1 and 2.2 mg l(-1)) daily for 14 days. The results obtained show that serum P(i) and ALP concentrations were elevated by increasing Cd(+2) concentration in water containing fish whereas serum Ca level was decreased. At the same time, the protective role of waterborne zinc (Zn(+2), 1 mg l(-1)) on the same parameters was also investigated. Results showing that daily treatment of fish with Zn(+2), increased the concentrations of Ca and ALP in serum by 2.07 and 1.86 fold and decreased serum P(i) level by 57.7% in comparison with Cd(+2) treatment. The combination of Cd(+2) and Zn(+2) on the same parameters was studied next. There was a significant (P < 0.05) elevation in serum Ca and ALP levels in comparison with Cd(+2) treatment. Decreasing in serum P(i) level was not significant in comparison with Cd(+2) treatments. The protective effect of Zn(+2) on Cd(+2) disturbances in serum parameters related to bone metabolism in this manuscript has been also discussed.

Comparative in vitro study of the intestinal absorption of titanium and iron in rats.

Rat Everted Gut Sac (EGS) model was employed to study the intestinal uptake of titanium and iron. Incubation of freshly prepared rat EGS in Earle's medium pH = 7.4 containing titanium showed that the absorption of titanium as well as iron was a dose dependent process. Ascorbic acid enhanced the absorption of both metal ions, while NaF (1 mM) as an inhibitor of glycolytic energy supply, decreased their absorption. The Na+-K+ ATPase inhibitor, ouabain (1 mM) reduced intestinal absorption of Titanium. This suggests that titanium uptake is an active transport process as is iron uptake. Iron absorption was reduced approximate by 17% when titanium was presented to incubation medium EGS whereas, the absorption of titanium was decreased by 35% when iron was added to the reaction mixture.

Effect of in vitro exposure to Vibrio vulnificus on hydroelectrolytic transport and structural changes of sea bream (Sparus aurata L.) intestine.

The everted gut sac technique has been used to investigate the effect of Vibrio vulnificus on water and electrolyte (Na(+), K(+), Cl(-), HCO(3)(-)) transport on the intestine of sea bream (Sparus aurata L.). Both the anterior and the posterior intestine were incubated in a medium containing 10(8) V. vulnificus cells ml(-1) at 25 degrees C for 2 h. The presence of V. vulnificus resulted in a significant reduction (P < 0.05) of water absorption in the anterior intestine, while sodium absorption in the anterior (P < 0.01) and posterior (P < 0.05) intestine was elevated. Chloride absorption was increased, but the changed was not significant, while potassium absorption decreased significantly (P < 0.05), but only in the posterior intestine. Incubation the sea bream intestine with V. vulnificus did not affect carbonate secretion in the anterior segment, whereas high secretion was stimulated in the posterior segment (P < 0.01). Histological evaluations demonstrated damage in the anterior intestine of sea bream that was characterized by the detachment of degenerative enterocytes, alterations in the microvilli, and the presence of a heterogenous cell population, indicating inflammation. Based on our results, we conclude that V. vulnificus caused cell damage to the intestine of sea bream and that the anterior intestine is more susceptible than the posterior part of the intestine. Several hypotheses are suggested to explain our observations, such as the presence of higher numbers of villosities in the anterior intestine than in the posterior one and/or the presence of endogenous bacteria in the posterior intestine which may have a protector role.

Comparative effects of copper, iron, vanadium and titanium on low density lipoprotein oxidation in vitro.

INTRODUCTION: Oxidation of low density lipoprotein (LDL) has been strongly implicated in the phathogenesis of atherosclerosis. The use of oxidants in dietary food stuff may lead to the production of oxidized LDL and may increase both the development and the progression of atherosclerosis. The present work investigated the effects of some elements including: copper (Cu), iron (Fe), vanadium (V) and titanium (Ti) on in vitro LDL oxidation quantitatively. METHODS: The first LDL fraction was isolated from fresh plasma by single vertical discontinuous density gradient ultracentrifugation. The formation of conjugated dienes and thiobarbituric acid reactive substances and increase in electrophoretic mobility of LDL were monitored as markers of the oxidation of LDL. RESULTS: It was demonstrated that Cu, Fe, V and Ti exhibited strong oxidant activity in this respect (P<0.001). Oxidation of LDL in the presence of Cu was more and appeared to be in this order Cu>Fe>V>Ti. DISCUSSION: Cu, Fe, V and Ti are redox-active transition metals that may cause oxidative damage to lipids, proteins and DNA molecules. We suggest that these elements may also influence the oxidation of LDL in vivo, which could increase both the development and progression of atherosclerosis.