Ameliorative Effects of Nilotinib on CCl4 Induced Liver Fibrosis Via Attenuation of RAGE/HMGB1 Gene Expression and Oxidative Stress in Rat / 전남의대학술지

Nilotinib as a tyrosine kinase inhibitor has been recently used to improve the liver fibrosis process, but the exact mechanisms still require further clarification. In this study, we investigated the anti-fibrotic effects of Nilotinib via RAGE/HMGB1axis and antioxidant mechanisms. This experimental study was performed in the Hamadan University of Medical Sciences, Iran, from May 2015 to December 2016. Liver fibrosis was induced in Wistar male rats by CCL₄. Rats were gavaged daily with Nilotinib (10 mg/kg). RAGE, HMGB1, TNF-α and TGF-β mRNA expression were evaluated by quantitative RT-PCR. TNF-α protein levels were measured using the immunoassay method. Thiol groups, carbonyl groups, nitric oxide levels and glutathione peroxidase activity were measured by spectrophotometric methods.The results showed that Nilotinib decreased TNF-α, TGF-β, RAGE and HMGB1 mRNA expression (p<0.001) in the liver tissues of the fibrosis group. Nilotinib also decreased carbonyl groups and nitric oxide levels and increased thiol groups and glutathione peroxidase activity in the fibrosis groups. The histopathological changes were found to be attenuated by Nilotinib. In conclusion, Nilotinib can improve liver fibrosis and open new mechanisms of the anti-fibrotic properties of Nilotinib.

Ameliorative Effects of Nilotinib on CCl4 Induced Liver Fibrosis Via Attenuation of RAGE/HMGB1 Gene Expression and Oxidative Stress in Rat / 전남의대학술지

Nilotinib as a tyrosine kinase inhibitor has been recently used to improve the liver fibrosis process, but the exact mechanisms still require further clarification. In this study, we investigated the anti-fibrotic effects of Nilotinib via RAGE/HMGB1axis and antioxidant mechanisms. This experimental study was performed in the Hamadan University of Medical Sciences, Iran, from May 2015 to December 2016. Liver fibrosis was induced in Wistar male rats by CCL₄. Rats were gavaged daily with Nilotinib (10 mg/kg). RAGE, HMGB1, TNF-α and TGF-β mRNA expression were evaluated by quantitative RT-PCR. TNF-α protein levels were measured using the immunoassay method. Thiol groups, carbonyl groups, nitric oxide levels and glutathione peroxidase activity were measured by spectrophotometric methods.The results showed that Nilotinib decreased TNF-α, TGF-β, RAGE and HMGB1 mRNA expression (p<0.001) in the liver tissues of the fibrosis group. Nilotinib also decreased carbonyl groups and nitric oxide levels and increased thiol groups and glutathione peroxidase activity in the fibrosis groups. The histopathological changes were found to be attenuated by Nilotinib. In conclusion, Nilotinib can improve liver fibrosis and open new mechanisms of the anti-fibrotic properties of Nilotinib.

Peroxisome Proliferator-Activated Receptor-γGene Expression and Its Association with Oxidative Stress in Patients with Metabolic Syndrome / 전남의대학술지

Regulation of the peroxisome proliferator-activated receptor-γ (PPAR-γ) gene plays an important role in controlling the metabolism of lipids and inflammatory processes. Therefore, it can be associated with the pathogenesis of metabolic syndrome (MetS). The purpose of this study was to determine the expression of this gene in peripheral blood mononuclear cells (PBMC) in patients with metabolic syndrome. Using real-time polymerase chain reaction (PCR), mRNA expression of PPAR-γ was found in PBMC from 37 subjects with MetS and 30 healthy controls. Serum levels of glucose and lipid profiles were measured. The total antioxidant capacity (TAC) was measured using the ferric reducing ability of plasma (FRAP) test. Malondialdehyde (MDA) was determined using a fluorimetric method. Total oxidant status (TOS) in serum was assayed according to oxidation of ferric to ferrous in the presence of methyl orange. Super oxide dismutase (SOD) activity was measured using a Randox kit. Expression of PPAR-γ gene was significantly increased in patients with MetS compared to the control subjects (p=0.002). There was no difference in serum levels of TAC, MDA and SOD between the two study groups, but a significant difference was observed in the TOS (p=0.03). Serum levels of triglycerides and glucose were significantly higher in subjects with MetS. According to the results of our study, an increase in the expression of PPAR-γ in subjects with MetS indicated a possible role of PPAR-γ in the pathogenesis of this disease.

Peroxisome Proliferator-Activated Receptor-γGene Expression and Its Association with Oxidative Stress in Patients with Metabolic Syndrome / 전남의대학술지

Regulation of the peroxisome proliferator-activated receptor-γ (PPAR-γ) gene plays an important role in controlling the metabolism of lipids and inflammatory processes. Therefore, it can be associated with the pathogenesis of metabolic syndrome (MetS). The purpose of this study was to determine the expression of this gene in peripheral blood mononuclear cells (PBMC) in patients with metabolic syndrome. Using real-time polymerase chain reaction (PCR), mRNA expression of PPAR-γ was found in PBMC from 37 subjects with MetS and 30 healthy controls. Serum levels of glucose and lipid profiles were measured. The total antioxidant capacity (TAC) was measured using the ferric reducing ability of plasma (FRAP) test. Malondialdehyde (MDA) was determined using a fluorimetric method. Total oxidant status (TOS) in serum was assayed according to oxidation of ferric to ferrous in the presence of methyl orange. Super oxide dismutase (SOD) activity was measured using a Randox kit. Expression of PPAR-γ gene was significantly increased in patients with MetS compared to the control subjects (p=0.002). There was no difference in serum levels of TAC, MDA and SOD between the two study groups, but a significant difference was observed in the TOS (p=0.03). Serum levels of triglycerides and glucose were significantly higher in subjects with MetS. According to the results of our study, an increase in the expression of PPAR-γ in subjects with MetS indicated a possible role of PPAR-γ in the pathogenesis of this disease.

Dill tablet: A potential antioxidant and anti-diabetic medicine

Objective: To evaluate the antiglycation and antioxidant properties of the dill tablet, an herbal product used in Iran as a hypolipidemic medicine. Methods: In this descriptive study, the antioxidant and antiradical properties of dill tablet at different concentration (0.032, 0.065, 0.125, 0.25, 0.5 and 1 mg/mL) were measured. The total phenolic, flavonols and flavonoid, alkaloids, anthocyanin, tannin and saponin contents in dill tablet were determined. Furthermore, antiglycation properties of dill tablet were assayed. In the in vivo experiments, male rats were randomly divided into three groups (n = 6): Group 1: normal rats; Group 2: diabetic rats; Group 3: diabetic rats + 300 mg/kg dill tablet, and Group 4: diabetic rats + 100 mg/kg dill tablet. After 2 months, the blood glucose was measured enzymatically and advanced glycation end-products (AGEs) formation was determined using a fluorometric method. Results: Our results illustrated that different concentrations of dill tablet had significant antioxidant activity. Dill tablet markedly declined AGEs formation and fructosamine levels (P < 0.001) compared with glycated sample. Oxidation of protein carbonyl and thiol group was significantly reduced by dill tablet in a dose dependent manner (P < 0.001). Formation of amyloid cross-β and fragmentation were markedly inhibited by dill tablet (P < 0.001) compared with glycated sample. After 2 months, fasting blood glucose levels (P < 0.001) and AGEs formation (P < 0.05) were significantly reduced by dill tablet in diabetic animals. Conclusions: Dill tablet exhibited significant antiglycation and antioxidant activities. This study provides a scientific basis for using dill in treatment of diabetic patients.

Serum levels of lycopene, beta-carotene, and retinol and their correlation with sperm DNA damage in normospermic and infertile men

Background: Oxidative stress in reproductive system leads to sperm DNA damage and sperm membrane lipid peroxidation and may play an important role in the pathogenesis of male infertility, especially in idiopathic cases. Antioxidants such as carotenoids function against free radical damages

Objective: The aim of this study was to determine the levels of lycopene, beta-carotene and retinol in serum and their relationship with sperm DNA damage and lipid peroxidation in infertile and normospermic males

Materials and Methods: Sixty two infertile men and 71 normospermic men participated in this study. Blood and semen samples were collected from all subjects. Sperm DNA damage was measured using TUNEL method. Carotenoids, retinol, and malonedildehyde in serum were also determined

Results: DNA fragmentation was higher in infertile group comparing to control group. Serum levels of lycopene, beta-carotene and, vitamin A in infertile men were significantly lower than normospermic men [p< 0.001, =0.005, and =0.003 respectively]. While serum MDA was not significantly different between two groups, MDA in seminal plasma of infertile men was significantly higher than control group [p< 0.001]

Conclusion: We concluded that lycopene, beta-carotene, and retinol can reduce sperm DNA fragmentation and lipid peroxidation through their antioxidant effect. Therefore the DNA fragmentation assay and determination of antioxidants factors such as lycopene, beta-carotene and retinol, along with sperm analysis can be useful in diagnosis and treatment of men with idiopathic infertility

Body fat and plasma lipid profile in different levels of physical fitness in male students

Adiposity is correlated with metabolic complications, such as insulin resistance, glucose intolerance, hypertension etc. that finally can lead to coronary heart disease [CHD]. Physical activity plays an important role in prevention of these diseases. We aimed to study body fat and plasma lipids and lipoproteins in different levels of physical fitness; also to examine any relationship between body fat and level of physical fitness. In this cross-sectional study carried out in Hamadan University of Medical Sciences [Hamadan, Iran] during 2010-2011, 80 healthy volunteer male students [19-27 yr old] were enrolled. The subjects were divided to three groups i.e. good, average and weak according to their physical fitness scores. Their lipid profile was determined using an auto-analyzer and commercial enzymatic spectorophptometric kits. Besides, body fat was determined using skinfold technique. The fitness tests were carried out for all participants, including: speed, stamina, aerobic endurance, muscular strength etc. The mean of body fat [%] in the subjects with good, medium and weak levels of physical activity was 8.25, 11.98, and 22.65% respectively. There were no significant differences in lipid parameters between the subjects with different level of physical fitness. A negative significant correlation was observed between physical fitness and body fat [r=-0.27, P=0.034]. The body fat increased with increases in serum triglyceride. The presence of difference in mean of body fat percent in different physical fitness confirms that aerobic activity improves the plasma lipid and lipoprotein profile, and can diminish the body fat

Effects of treatment with platinum azidothymidine and azidothymidine on telomerase activity and Bcl-2 concentration in hepatocellular carcinoma- induced rats

Telomerase activity increases in cancer cells. Bcl-2 is an antiapoptotic factor that its concentration grows in many cancer cells including hepatocellular carcinoma cells. In this study, an attempt was made to investigate the effects of a new synthetic compound, platinum azidothymidine [Pt-AZT] on treatment of rats with Hepatocellular Carcinoma [HCC] and to compare its effects with azidothymidine [AZT] in alteration of telomerase activity and Bcl-2 concentration in HCC. Healthy adult male Wistar rats [n=100] were randomly divided into 4 groups [A, B, C, and D]. Group A contained 25 healthy rats and was considered as the control group. Liver preneoplastic lesions were induced in remaining animals [n=75] using Solt-Farber resistant hepatocyte protocol. These animals were randomly allocated in groups B, C and D. Group B was negative control [untreated], groups C and D were treated by intraperitoneal injection [IP] of Pt-AZT [0.9 mg/kg/day] and AZT [0.3 mg/kg/day], respectively for 14 days. After the treatment period, telomerase activity and Bcl-2 concentration were determined in the rats' liver. No HCC was developed in group A, but tumors were present in all other groups. Telomerase activity and Bcl-2 concentration were significantly lower in group C compared to groups B [0.159 +/- 0.06 vs. 0.577 +/- 0.116 IU/L, p<0.001, respectively and 0.931 +/- 0.388 vs. 3.94 +/- 0.74 ng/ml, p<0.001, respectively]. Similar results were observed in comparison with group D [0.331 +/- 0.06 vs. 0.577 +/- 0.116 IU/L, p<0.001, respectively and 0.931 +/- 0.388 vs. 2.94 +/- 0.594 ng/ml, respectively]. There was a significant negative correlation between telomerase activity and Bcl-2 concentration only in untreated cancer group [p=0.034]. In this study, higher anticancer activity of Pt-AZT in comparison to AZT was demonstrated. It effectively inhibits the growth of liver tumor in rats through extending apoptosis

Levels of Salivary Antioxidant Vitamins and Lipid Peroxidation in Patients with Oral Lichen Planus and Healthy Individuals / 전남의대학술지

Oral lichen planus (OLP) is a chronic inflammatory mucosal disease of unknown etiology. Many studies have implicated the protective role of antioxidants in such diseases. The aim of this study was to compare salivary total antioxidant capacity (TAC and malondialdehyde (MDA) and antioxidant vitamin (vitamin s A, C and E) levels in patients with erosive OLP and healthy individuals. Thirty six patients with OLP (14 males, 22 females) and 36 control subjects (15 males, 21 females), matched for age and sex were enrolled in this case control study. The salivary levels of MDA, TAC, and antioxidant vitamin levels were measured in both case and control groups. The salivary level of MDA was significantly higher (p<0.001) in patients than in controls. In patients with OLP, the TAC of saliva was significantly lower than that in healthy subjects (p<0.001). Compared with controls, the levels of salivary antioxidant vitamins were significantly decreased in patients with OLP (p<0.001). In addition, a positive correlation was found between the decrease in the salivary amount of vitamin C and that in vitamin E in patients and controls. In addition to the lower salivary levels of antioxidant vitamins and the lower TAC, the higher level of MDA in patients with OLP suggests that free radicals and the resulting oxidative damage may be important in the pathogenesis of OLP lesions.

Levels of Salivary Antioxidant Vitamins and Lipid Peroxidation in Patients with Oral Lichen Planus and Healthy Individuals / 전남의대학술지

Oral lichen planus (OLP) is a chronic inflammatory mucosal disease of unknown etiology. Many studies have implicated the protective role of antioxidants in such diseases. The aim of this study was to compare salivary total antioxidant capacity (TAC and malondialdehyde (MDA) and antioxidant vitamin (vitamin s A, C and E) levels in patients with erosive OLP and healthy individuals. Thirty six patients with OLP (14 males, 22 females) and 36 control subjects (15 males, 21 females), matched for age and sex were enrolled in this case control study. The salivary levels of MDA, TAC, and antioxidant vitamin levels were measured in both case and control groups. The salivary level of MDA was significantly higher (p<0.001) in patients than in controls. In patients with OLP, the TAC of saliva was significantly lower than that in healthy subjects (p<0.001). Compared with controls, the levels of salivary antioxidant vitamins were significantly decreased in patients with OLP (p<0.001). In addition, a positive correlation was found between the decrease in the salivary amount of vitamin C and that in vitamin E in patients and controls. In addition to the lower salivary levels of antioxidant vitamins and the lower TAC, the higher level of MDA in patients with OLP suggests that free radicals and the resulting oxidative damage may be important in the pathogenesis of OLP lesions.

Effects of ascorbic acid on serum level of unconjugated estriol and its relationship with preterm premature rupture of membrane: a double-blind randomized controlled clinical trial

Background: Vitamin C is a water-soluble antioxidant that not only stimulates and protects collagen synthesis but also plays an important role in maintaining cellular integrity in a normal pregnancy. This study surveyed the effects of ascorbic acid on the serum level of unconjugated estriol and the relationship between unconjugated estriol and preterm premature rupture of membrane [PPROM]

Methods: This double-blind, randomized clinical trial recruited 60 patients with predisposing factors to PPROM. The women were randomly divided into two groups of intervention and control and received vitamin C and placebo, respectively. The intervention group received 250 mg vitamin C twice a day and the controls received the placebo only. Unconjugated estriol was measured using the ELISA. All data were extracted and recorded in a checklist and compared using descriptive statistics as well as the chi[2], Fisher exact, and t tests

Results: The demographic data showed no difference between the two groups. The mean level of serum unconjugated estriol was significantly lower in the intervention group than in the control group [P=0.044]. Also, the frequency of PPROM was lower in the intervention group, but the difference was not significant [P>0.05]. Unconjugated estriol levels were not significantly different between the healthy women and the PPROM patients

Conclusion: This study demonstrated that vitamin C administration decreased unconjugated estriol levels in the patients with PPROM. The findings of this study also indicated that administration of ascorbic acid was a safe and effective method to reduce the incidence of PPROM. Alteration in unconjugated estriol is an active mediator for this effect

The relationship between the level of salivary alpha amylase activity and pain severity in patients with symptomatic irreversible pulpitis

OBJECTIVES: Assessment of dental pain severity is very challenging in dentistry. Previous studies have suggested that elevated salivary alpha amylase may contribute to increased physical stresses. There is a close association between salivary alpha amylase and plasma norepinephrine under stressful physical conditions. The aim of this study was to evaluate the relationship between pain severity and salivary alpha amylase levels in patients with symptomatic irreversible pulpitis. MATERIALS AND METHODS: Thirty-six patients (20 females and 16 males) with severe tooth pain due to symptomatic irreversible pulpitis were selected. The visual analogue scale (VAS) score was used to assess the pain severity in each patient. Unstimulated whole saliva was collected, and the level of alpha amylase activity was assessed by the spectrophotometric method. Statistical analysis was performed using SPSS 13. RESULTS: The level of alpha amylase was significantly increased in the saliva in association with pain severity assessed by VAS. The salivary alpha amylase was also elevated with increased age and in males. CONCLUSIONS: There was a significant correlation between the VAS pain scale and salivary alpha amylase level, which indicates this biomarker may be a good index for the objective assessment of pain intensity.

Study of nonenzymatic glycation of transferring and its effect on iron-binding antioxidant capacity

Nonenzymatic glycosylation [glycation] occurs in many macromolecules in aging and diabetes due to exposure of biomolecules to high level of glucose. Glycation can changes function, activities and structure of many biomolecules. Considering this important role of transferring [Trf] in iron transport and antioxidant activity in plasma this study was carried out to investigate the effect of glycation in these processes. In this study, human pro-Trf [5 mg/ml in sodium phosphate buffer pH= 7.4] was treated with different concentrations of glucose in different period of times [10 days and 20 days]. Rate of glycation was measured using thiobarbituric acid method. The effect of glycation on iron binding antioxidant capacity of apo-Trf was investigated using two methods [RBC hemolysis and fluorescent]. Result showed that rate of glycation of apo-Trf was increased with increase in glucose concentration and time of incubation [P< 0.05]. Lower iron binding antioxidant capacity was observed for glycted Trf as compared to native Trf [P< 0.05]. Impairment of antioxidant capacity of glycated Trf can suggest a relationship between glycation of Trf and oxidative stress that occurs due to hyperglycemia in diabetic patients

[Correlation between serum aldosterone level and hearing condition of elderly patients referred to otolaryngology services of Hamadan, Western Iran]

Recently, more attention was paid to the direct protective effect of aldosterone against hearing impairment in elderly patients. The aim of this study was determination of possible correlation between serum aldosterone level and hearing condition of elderly patients that referred to the Otolaryngology services of Hamadan in 2005-2006. In this case control study 54 [27 males, 27 females] persons above 60 years old were evaluated. They contained twenty eight cases with normal hearing and 26 cases with presbycusis. Persons with any abnormal biochemical finding or history of conditions that predispose them to the sensorineural hearing loss [SNHL] were excluded. In both groups serum level of sodium, potassium and aldosterone were measured and hearing condition evaluated by puretone, speech and immitance audiometry. Statistical relationship between serum aldosterone level and hearing condition, sex, configuration of audiogram and speech discrimination score [SDS] were not significant. In addition, no significant relationship between sodium and potassium levels with hearing conditon was found [p>0.05]. This study could not confirm protective effect of aldostrone against prebycusis. This discrepancy may originate from epidemiologic differences, laboratory errors or small sample size

Cytotoxic effect of [glycated albumin-transition metal ion] on rat hepatocyte suspension

Combination of glycation and oxidation is associated with diabetes mellitus. The aim of this study was to clarify the effect of glycated proteins in presence of transition metal ions on production of reactive oxygen species [ROS] in rat hepatocyte suspension. Glycated albumin was prepared by incubation of bovine serum albumin with 100 mM glucose in 0.3 M phosphate buffer at 37°C for 2, 4, and 6 weeks. The prepared rat liver cell suspension was treated with glycated albumin in presence of either Fe[+++] or Cu[++] ions. Produced malone-dialdehyde was measured as an indicator of ROS and of cell injury. The results showed Fe[+++] and Cu[++] ions increase the ROS production in presence of glycated albumin [p<0.01]. All prepared glycated albumin showed cytotoxicity in rat hepatocytes suspension in the presence of cupric and ferric ions, and this injury was dependent to metal ion concentration. Higher degree of glycation showed higher effect on ROS production [P<0.01] Comparing the effect of Fe[+++] and Cu[++], cupric ion had higher cytotoxic effect [p<0.01]. Conclusion: These results indicated that hepatocytes may be damaged by ROS which are produced by the interaction of the glycated albumin and transition metal ion

Glycation of human IgG induces structural alterations leading to changes in its interaction with Anti-IgG

Glycation of proteins is a non-enzymatic spontaneous process that occurs in diabetes mellitus and aging, altering the structure and function of proteins. IgG undergoes glycation leading to changes in its reactivity to antigen and fixation of complement. This study aimed at revealing the effect of glycation on the interaction of IgG with anti-IgG using electroimmunoassay. Purified human IgG was glycated with different concentrations of glucose and different periods of treatment. Glycation was measured using thiobarbituric acid reaction. Glycated and non-glycated IgG were subjected to electroimmunoassay, and the height of the precipitated rings were measured and compared. The results showed that IgG was glycated in vitro and the level of glycation was dependent on the glucose concentration and duration of treatment with glucose. The height of glycated IgG peaks formed in the electroimmunoassay was significantly lower than those of non-glycated IgG [p < 0.01]. The results indicated that in vitro glycation of IgG leads to structural changes altering its mobility in the electroimmunoassay. Moreover, it suggests that this alteration may cause the weakness of its interaction with anti-IgG. This phenomenon may play a role in the susceptibility of diabetic patients to infection

In vitro glycation of human IgG and its effect on interaction with anti-IgG

Non-enzymatic glycosylation of proteins is one of the key mechanisms in the pathogenesis of diabetic complications. Glycation of IgG is of special interest due to its possible influence on the functionality of immunoglobulins and overall immunocompetence. The aim of this study was to clarify more details of in vitro glycation of IgG and to study the effect of this modification on its interation with anti-IgG. Purified human IgG was glycated in the presence of 50 and 100 mM glucose. Glycation was measured using spectrophotometric thiobarbituric acid method. To study the effect of glycation on interaction with anti IgG the Single Radial Immunodiffusion [SRID] was used and the diameters of precipitation rings of glycated IgG and non-glycated IgG were measured and compared.The results showed that IgG was glycated in presence of 50 and 100 mM glucose at 27[degree] and 37[degree] C and the extent of glycation was dependent on glucose concentration and time of incubation. In higher concentration of glucose and longer period of incubation glycation was higher at 27[degree] C [p<0.01]. Similar results were obtained at 37[degree] C.The results of SRID indicated that glycated IgG showed reduced interaction with anti-IgG. The diameters of precipitated rings for glycated IgG were significantly lower than those of non-glycated IgG [p < 0.01].It can be concluded that modification that occurred in IgG structure due to glycation can be the reason of the reduction of its interaction with anti-IgG