Sulfhydryl Levels Research Articles

ОКИСНА МОДИФІКАЦІЯ БІЛКІВ У СИРОВАТЦІ КРОВІ ЩУРІВ В УМОВАХ КАРРАГІНАН-ІНДУКОВАНОГО ЗАПАЛЕННЯ ЗАДНЬОЇ КІНЦІВКИ ТА ТРИВАЛОГО ПРОФІЛАКТИЧНОГО ВВЕДЕННЯ ХОНДРОЇТИНУ СУЛЬФАТУ

The aim of the work was to investigate the preventive effect of Chondroitin Sulfate on the content of products of oxidative modification of proteins and the level of sulfhydryl groups in blood serum of rats at local inflammation of the hind limb. The studies were conducted on white non-linear, sexually mature male rats weighing 180–240 g, in compliance with the general ethical principles of experiments on animals. All animals were divided into four experimental groups. The first group – control: animals sub-planar injected 0,1 ml of 0,9 % NaCl solution into the posterior right limb. The second group – animals received a therapeutic dose of 3 mg x kg-1 chondroitin sulfate daily for 28 days daily. The third group – animals were infused intramuscularly with 0,1 ml of 0,9 % NaCl solution in the posterior right limb for 28 days and for 29 days inflammatory edema of the limb was stimulated (animals were sub-planar injected with 0,1 ml of 1 % carrageenan solution to the posterior right limb ) The fourth group – for 28 days rats were daily intramuscularly injected with a therapeutic dose of 3 mg x kg-1 chondroitin sulfate, after which on 29th day, inflammatory edema of the limb was stimulated. Animals were killed 3 hours after injection of carrageenan solution according to the protocol of the ethical committee, and then blood sampling for further research was quickly taken. The total number of animals involved in experimental studies was 40 individuals. The content of the products of oxidative modification of proteins (OMP) and oligopeptides was determined by the level of carbonyl derivatives that were detected in reaction with 2,4-dinitrophenylhydrazine. The level of total, protein-bound and non-protein sulfhydryl (SH) -groups was measured by the Elman method. It has been established that with carrageenan-induced inflammation of the posterior limb, the content of the products of oxidative modification of proteins increases and the content of sulfhydryl groups decreases in the serum. It was shown that the prophylactic administration of chondroitin sulfate based drug on animals with carrageenan-induced inflammation restored the abovementioned parameters.

Plasma antioxidant enzyme levels in smoker and non-smoker pregnant women and their correlation with uterine arterial Doppler parameters

Introduction: Cigarettes contain many free radicals and cause oxidative stress in the body. Its contents decrease blood fluidity and blood stream by affecting the vascular endothelium; further, they directly or indirectly impair placental perfusion. Material and Method: Catalase (CAT), malondialdehyde (MDA) and total sulfhydryl (SH) levels were analysed in venous blood specimens obtained from 49 smoker and 35 non-smoker pregnant women (age, 18-35 y) who were in the second trimester. Uterine arterial Doppler ultrasonography was performed to evaluate uterine perfusion. Pulsatile index (PI), resistance index (RI) and systole/diastole ratio (S/D) was measured. The smoker group was further subdivided into two groups: 1. those who smoked for 5 y. Obtained data were analysed using the SPSS 11 statistical programme. Results: No significant differences were observed between the smoker and non-smoker groups in terms of demographic parameters and right–left uterine arterial Doppler parameters. The mean S/D ratio was significantly higher in the smoker group (P=0.01). The mean PI, mean RI and plasma MDA, CAT and total SH levels were similar in both groups. Although not statistically significant, the mean PI, RI and S/D ratio values of the smoker pregnant women who smoked for ≥ 5 y was found to be higher than of those who smoked for <5 y. Conclusion: There is no correlation between plasma antioxidant enzyme levels and uterine artery Doppler parameters; however, the duration of smoking may affect uterine perfusion.

Mn Inhibits GSH Synthesis via Downregulation of Neuronal EAAC1 and Astrocytic xCT to Cause Oxidative Damage in the Striatum of Mice.

Excessive manganese (Mn) can accumulate in the striatum of the brain following overexposure. Oxidative stress is a well-recognized mechanism in Mn-induced neurotoxicity. It has been proven that glutathione (GSH) depletion is a key factor in oxidative damage during Mn exposure. However, no study has focused on the dysfunction of GSH synthesis-induced oxidative stress in the brain during Mn exposure. The objective of the present study was to explore the mechanism of Mn disruption of GSH synthesis via EAAC1 and xCT in vitro and in vivo. Primary neurons and astrocytes were cultured and treated with different doses of Mn to observe the state of cells and levels of GSH and reactive oxygen species (ROS) and measure mRNA and protein expression of EAAC1 and xCT. Mice were randomly divided into seven groups, which received saline, 12.5, 25, and 50 mg/kg MnCl2, 500 mg/kg AAH (EAAC1 inhibitor) + 50 mg/kg MnCl2, 75 mg/kg SSZ (xCT inhibitor) + 50 mg/kg MnCl2, and 100 mg/kg NAC (GSH rescuer) + 50 mg/kg MnCl2 once daily for two weeks. Then, levels of EAAC1, xCT, ROS, GSH, malondialdehyde (MDA), protein sulfhydryl, carbonyl, 8-hydroxy-2-deoxyguanosine (8-OHdG), and morphological and ultrastructural features in the striatum of mice were measured. Mn reduced protein levels, mRNA expression, and immunofluorescence intensity of EAAC1 and xCT. Mn also decreased the level of GSH, sulfhydryl, and increased ROS, MDA, 8-OHdG, and carbonyl in a dose-dependent manner. Injury-related pathological and ultrastructure changes in the striatum of mice were significantly present. In conclusion, excessive exposure to Mn disrupts GSH synthesis through inhibition of EAAC1 and xCT to trigger oxidative damage in the striatum.

31 - The radioprotective effect of caffeic acid phenethyl ester in the brain tissue of total head irradiated rats

In this study, we evaluated if caffeic acid phenethyl ester (CAPE) has protective effect on the damage in brain tissue induced by gamma radiation applied to rats, considering that may have inhibitory effects on ionizing radiation damage. To this end, 36 Male Wistar-Albino rats were used in our study. Our study groups are consisted of 4 subgroups: 2 control, radiotherapy (R), and KAFE + R. All of the groups except the control received a single dose of 5 Gy of radiotherapy on the first day. On day 11, brain tissues of all rats were removed and homogenized in phosphate buffer. Total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), paraoxanase (PON), arylesterase (ARE), ceruloplasmin (SER) and total sulfhydryl (SH) parameters were measured to determine if CAFE has protective effect. In conclusion, ARE, PON activities and total SH levels were statistically increased compared to the group R and LOOH, TOS and OSI levels significantly decreased. In conclusion, our findings show that CAFE might be having antioxidative effects to oxidative damage induced by radiation in brain of rats. However, to be confirmed these findings, it is needed to be supported by pharmacological and toxicological studies.

Phytochemical characterization of the Vochysia rufa (Vochysiaceae) extract and its effects on oxidative stress in the pancreata of streptozotocin-induced diabetic rats.

Aqueous extract of macerated Vochysia rufa stem bark has been commonly used in the treatment of diabetes. Therefore, we evaluated the antihyperglycemic and antioxidant effects of an extract of V. rufa on the pancreata of streptozotocin (STZ)-induced diabetic rats. Animals received one of the following treatments daily by oral gavage: water (diabetic-control), V. rufa extract (diabetic-V. rufa), or glibenclamide (diabetic-GBD). Total antioxidant capacity; levels of thiobarbituric acid reactive substances, reduced glutathione, and sulfhydryls; and superoxide dismutase, catalase, and glutathione peroxidase (GPx) activities were measured in the pancreas. Biochemical analysis of serum total cholesterol and fractions, triglycerides, creatinine, urea, acid uric, ALP, γ-GT, AST, and ALT was performed, and pancreatic β-cells positive for insulin were evaluated by immunohistochemistry. Rats treated with extract exhibited a decrease in fasting blood glucose compared with levels in diabetic control rats. GPx activity and sulfhydryl levels were significantly lower in diabetic-V. rufa rats compared with those of diabetic-control rats. V. rufa extract acted to normalize the biochemical alterations found in diabetic rats (diabetic-controls), as demonstrated by increases in urea, HDL, ALP, AST, and ALT. Reduction in blood glucose was independent of an increase in insulin. The V. rufa extract was found to be composed of free sugars (inositol, galactose, glucose, mannose, sucrose, arabinose, and ribose) as the main metabolites. Thus, aqueous extract of the stem bark of V. rufa is capable of reducing blood glucose, resulting in an antioxidant effect on the pancreatic tissue of STZ-diabetic rats.

Vitamin C as a modulator of oxidative stress in erythrocytes of stored blood

AimTo determine the effects of Vitamin C (VC-ascorbic acid) as an additive on erythrocytes of stored blood. BackgroundOxidative stress (OS) plays a major role in the formation of storage lesion of erythrocytes. Antioxidants, such as VC could be beneficial in combating oxidative damage during storage. Materials and methodsBlood obtained from male Wistar rats was stored at 4°C in anticoagulant solution citrate-phosphate-dextrose-adenine solution. Blood samples were divided into 3 groups – (i) Controls, (ii) VC 10 (VC at a concentration of 10mM), (iii) VC 30 (VC at a concentration of 30mM). Markers of OS in erythrocytes such as – hemoglobin, superoxides, antioxidant enzymes (superoxide dismutase; SOD, catalase and glutathione peroxidase), hemolysis, lipid peroxidation products (conjugate dienes and malondialdehyde), protein oxidation products and ascorbic acid were determined on days 0, 10 and 15 of storage. ResultsAddition of ascorbic acid to the storage solution contributed to the protection of erythrocytes from oxidative damage. Ascorbic acid at a concentration of 30mM decreased SOD levels and increased protein sulfhydryls (P-SH) levels on day 15 showing that higher concentration supplemented the inherent antioxidant defense system of erythrocytes during blood storage. ConclusionVC proved to be effective in combating OS during blood storage. However, further exploration of antioxidants as additives and the erythrocyte storage lesion would result in better management of blood storage.

Evaluation of adiponectin and leptin levels and oxidative stress in bipolar disorder patients with metabolic syndrome treated by valproic acid.

An increased risk for metabolic syndrome (MS) has been described for people with psychotic and mood disorders. The aim of this study was to determine the influence of valproic acid (VPA) treatment on adiponectin, leptin levels and oxidative stress in bipolar disorder (BD). Forty patients with BD receiving VPA monotherapy and 20 healthy control subjects were included in this study. BD patients were divided into two groups with and without MS as group 1 and group 2, respectively. Twenty BD patients diagnosed according to the Diagnostic and Statistical Manual for Mental Disorders (DSM IV) were assessed for MS according to the National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP III) criteria. Adiponectin, leptin, protein carbonyls, sulfhydryl (-SH) and malondialdehyde (MDA) levels were measured in 40 BD patients and 20 control subjects. Serum adiponectin levels were significantly lower in group 1 patients than in group 2 and control subjects ( p<.001). Serum leptin levels were significantly higher in group 1 patients than in group 2 and control subjects ( p<.001). Serum -SH levels were significantly lower in group 2 patients than in group 1 ( p<.001) and control subjects ( p<.05). Serum carbonyl levels were significantly higher in group 1 and group 2 patients than in control subjects ( p<.001). Serum MDA levels were significantly higher in group 1 patients than in group 2 and control subjects ( p<.001). These results provide further evidence that VPA treatment for patients with BD contributed to the metabolic disturbances, such as the decreased serum adiponectin and -SH levels, as well as the increased serum leptin, MDA and carbonyl levels.

Prevention of selenite‐induced cataractogenesis by sildenafil in rats

PurposeTo evaluate the effect of sildenafil on selenite‐induced cataract formation in a rat model.MethodsTwenty‐six Wistar rat pups were divided into four groups. Seven pups received only selenite on postpartum day 10 (group 1), 7 pups received selenite and additional high dose sildenafil on postpartum day 10 (group 2), 6 pups received selenite and additional low dose sildenafil on postpartum day 10 (group 3), and 6 pups received only saline (group 4, control). All pups were examined for the presence of cataract under the microscope, starting from the day their eyes opened. Nitrite oxide metabolites, advanced oxidation protein products and total sulfhydryl levels were evaluated in both serum and lenticular samples.ResultsIn group 3, the extent of lens opacification was significantly less than that of selenite‐injected untreated rats (group 1) (p &lt; 0.05). None of the rats in group 4 developed any lens opacity. In lenticular samples, nitrite oxide metabolites level was statistically lower in group 3 compared to group 1 (p &lt; 0.05). In serum, advanced oxidation protein products and total sulfhydryl levels was statistically lower in group 3 compared to group 1 (p &lt; 0.05).ConclusionsLow dose sildenafil appears to inhibit selenite‐induced cataractogenesis in the rat model, and this seems to be caused by the prevention of oxidative damage.

The structural damages of lens crystallins induced by peroxynitrite and methylglyoxal, two causative players in diabetic complications and preventive role of lens antioxidant components

Peroxynitrite (PON) and methylglyoxal (MGO), two diabetes-associated compounds, are believed to be important causative players in development of diabetic cataracts. In the current study, different spectroscopic methods, gel electrophoresis, lens culture and microscopic assessments were applied to examine the impact of individual, subsequent or simultaneous modification of lens crystallins with MGO and PON on their structure, oligomerization and aggregation. The protein modifications were confirmed with detection of the significantly increased quantity of carbonyl groups and decreased levels of sulfhydryl, tyrosine and tryptophan. Also, lens proteins modification with these chemical agents was accompanied with important structural alteration, oligomerization, disulfide/chromophore mediated protein crosslinking and important proteolytic instability. All these structural damages were more pronounced when the lens proteins were modified in the presence of both mentioned chemical agents, either in sequential or simultaneous manner. Ascorbic acid and glutathione, as the main components of lens antioxidant defense mechanism, were also capable to markedly prevent the damaging effects of PON and MGO on lens crystallins, as indicated by gel electrophoresis. The results of this study may highlight the importance of lens antioxidant defense system in protection of crystallins against the structural insults induced by PON and MGO during chronic hyperglycemia in the diabetic patients.

Safety protocol for the gold nanoparticles administration in rats

Gold nanoparticles (GNPs) have antioxidant and anti-inflammatory effects. However, toxicity is still a concern; therefore, it is critical to study both the therapeutic and toxic properties of GNPs. In this study, we evaluated the effects of the intraperitoneal administration of GNPs (20nm, at a concentration of 2.5mg/L for 21days) every 24 or 48h on oxidative stress, antioxidant status, and electron chain transport (ETC) in the brain. Liver histology and blood marker analyses were conducted to establish a time routine of GNP administration. The concentrations of GNP in the brain and liver were similar. Hepatic and serum levels of cholesterol, triglycerides, and transaminases were not altered after the administration of GNP every 24 or 48h. The superoxide and nitric oxide levels were unchanged after administration of GNP. Dichlorodihydrofluorescein (DCFH) levels decreased after the administration of GNP every 48h compared with that in the saline group. Sulfhydryl and carbonyl levels, as well as superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and glutathione (GSH) activities were not altered in the brain after administration of GNP in the two time periods studied. The GNP 48h group showed increased brain ETC activity. Compared to that in the saline group, the GNP 24h group showed marked parenchyma changes with cell necrosis and leukocyte infiltration. We therefore suggest that a concentration of 2.5mg/L of GNP administered every 48h has potential therapeutic benefits without toxicity.

Cytotoxic effects of cadmium and zinc co-exposure in PC12 cells and the underlying mechanism

Cadmium (Cd2+) is a well studied inducer of cellular necrosis and apoptosis. Zinc (Zn2+) is known to inhibit apoptosis induced by toxicants including Cd2+ both in vitro and in vivo. The mechanism of Zn2+-mediated protection from Cd2+-induced cytotoxicity is not established. In this study, we aimed to understand the effects of Zn2+ on Cd2+-induced cytotoxicity and apoptosis using PC12 cells. Cell viability and DNA fragmentation assays in PC12 cells exposed to Cd2+ and/or Zn2+ revealed that Cd2+ (5 and 10 μmol/L) alone induced significant cell death, and co-exposure to Zn2+ (5, 10, and 100 μmol/L) for 48 h had a protective effect. Assessment of intracellular free sulfhydryl levels and lactate dehydrogenase activity suggested that Cd2+ (10 μmol/L) induced oxidative stress and disrupted cell membrane integrity. Addition of Zn2+ (10 and 100 μmol/L) reduced Cd2+-mediated cytotoxicity. Changes in expression of the apoptotic factors Bax, Bcl-2, Bcl-x, and cytochrome c were measured via western blot and expression of caspase 9 was detected via reverse transcriptase polymerase chain reaction. Western blots showed that Zn2+ (10 and 100 μmol/L) suppressed Cd2+-induced apoptosis (10 μmol/L) by reducing cytochrome c release into the cytosol, and downregulating the proapoptotic protein, Bax. In addition, expression of caspase 9 was lower in Cd2+ (5 μmol/L)-treated PC12 cells when co-treated with Zn2+ (2 and 5 μmol/L). These findings suggest that the effective inhibition of Cd2+-induced apoptosis in PC12 cells by Zn2+ might be due to suppression of mitochondrial apoptosis pathway and inhibition of Cd2+-induced production of reactive oxygen species.

In vitro effect of N-acetyl-L-cysteine on glutathione and sulfhydryl levels in X-linked adrenoleukodystrophy patients

Introduction: Recent evidence shows that oxidative stress seems to be related with the pathophysiology of X-linked adrenoleukodystrophy (X-ALD), a neurodegenerative disorder. Methods: In the present study, the in vitro effect of N-acetyl-L-cysteine (NAC) on glutathione (GSH) and sulfhydryl levels in X-ALD patients was evaluated. Results: A significant reduction of GSH and sulfhydryl content was observed in X-ALD patients compared to the control group. Furthermore, 5 mM of NAC, in vitro, led to an increase in GSH content and sulfhydryl groups in these patients. Conclusion: These data probably indicate that an adjuvant therapy with the antioxidant NAC could improve the oxidative imbalance in X-ALD patients. Keywords: X- linked adrenoleukodystrophy; N-acetyl-L-cysteine; glutathione; sulfhydryl

Influence of L-carnitine on Stored Rat Blood: A study on Plasma

Objective: Plasma acts as a good indicator of oxidative stress in blood. L-Carnitine is an antioxidant that reduces metabolic stress in cells, thereby providing a protective effect against oxidative stress (OS). L-Carnitine as an additive in storage has not been explored. Thus, this study attempts to analyze the role of L-carnitine in blood storage solution, citrate phosphate dextrose adenine (CPDA)-1, through OS markers including antioxidant enzymes, lipid peroxidation, and protein oxidation.Materials and Methods:Blood was collected from male Wistar rats and stored in CPDA-1 solution with L-carnitine (10 mM, 30 mM, and 60 mM: groups LC 10, LC 30, and LC 60, respectively) and without L-carnitine (control group). Plasma was isolated every 5th day and the OS markers were analyzed.Results:Superoxide dismutase (SOD) and sulfhydryl (SH) increased over storage in controls, LC 30, and LC 60. Catalase increased in LC 30 and LC 60 during storage. Thiobarbituric acid reactive substances (TBARS) and protein carbonyl (PrC) levels in all groups increased initially and reduced towards the end of storage. SOD and SH levels were maintained while TBARS and PrC levels increased in LC 10.Conclusion:L-Carnitine was beneficial in terms of increased antioxidant capacity and SH and decreased lipid peroxidation. This forms the basis for further studies on L-carnitine as a constituent in storage solutions.

N-Acetyl-L-Cysteine Protects Liver and Kidney Against Chromium(VI)-Induced Oxidative Stress in Mice.

Acute hexavalent chromium [Cr(VI)] compound exposure may lead to hepatotoxic and nephrotoxic effects. Cr(VI) reduction may generate reactive intermediates and radicals which might be associated with damage. We investigated effects of N-acetyl-l-cysteine (NAC) pre- or post-treatment on oxidative stress and accumulation of Cr in liver and kidney of Cr(VI)-exposed mice. Intraperitoneal potassium dichromate injection (20mgCr/kg) caused a significant elevation of lipid peroxidation in both tissues as compared to control (p<0.05). Significant decreases in non-protein sulfhydryl (NPSH) level, as well as enzyme activities of catalase (CAT) and superoxide dismutase (SOD) along with significant accumulation of Cr in the tissues (p<0.05) were of note. NAC pre-treatment (200mg/kg, ip) provided a noticeable alleviation of lipid peroxidation (p<0.05) in both tissues, whereas post-treatment exerted significant effect only in kidney. Similarly, Cr(VI)-induced NPSH decline was restored by NAC pre-treatment in both tissues (p<0.05); however, NAC post-treatment could only replenish NPSH in liver (p<0.05). Regarding enzyme activities, in liver tissue NAC pre-treatment provided significant restoration on Cr(VI)-induced CAT inhibition (p<0.05), while SOD enzyme activity was regulated to some extent. In kidney, SOD activity was efficiently restored by both treatments (p<0.05), whereas CAT enzyme alteration could not be totally relieved. Additionally, NAC pre-treatment in both tissues and post-treatment in liver exerted significant tissue Cr level decreases (p<0.05). Overall, especially NAC pre-treatment seems to provide beneficial effects in regulating pro-oxidant/antioxidant balance and Cr accumulation caused by Cr(VI) in liver and kidney. This finding may be due to several mechanisms including extracellular reduction or chelation of Cr(VI) by readily available NAC.

Obesity is not a descriptive factor for oxidative stress and viscosity in follicular fluid of in vitro fertilization patients

Obesity's impact on micro-environmental oxidative stress and follicular fluid (FF) viscosity and whether or not it has any effect on in vitro fertilization (IVF) success is a matter of debate. In this study, our aim was to evaluate the levels of oxidative stress markers and the FF viscosity in obese and non-obese patients. Eighty norm-responder patients undergoing IVF were prospectively grouped according to their body mass indexes (BMI). Group 1 (n=40) and group 2 (n=40) had BMI values of ≤24.9 and ≥25.0, respectively. Total sulfhydryl (RSH) levels (nmol/m) and the formation of thiobarbituric acid-reactive substances (malondialdehyde, or MDA) (µmol/ml) in FFs were quantified. For the first time in our study, FF viscosity with changing BMI values was also determined. The mean levels of MDA (µmol/ml) and RSH (nmol/ml) were not significantly different between groups (1.37±0.51; 1.51±0.51; p>0.05 for MDA and 0.42±0.30; 0.41±0.20; p>0.05 for RSH, respectively). Similarly, the FF viscosity (centipoise) was not different between groups (1.28±0.28; 1.30±0.19; p<0.05, respectively). Independent of BMI, no correlation was found between FF levels of oxidative markers and the number of oocytes retrieved or the fertilization rates. In our study, we found no difference in the levels of follicular oxidative and anti-oxidative markers or the follicular fluid viscosity with changing BMI values. We also demonstrated that the levels of oxidative stress markers and the viscosity of follicular fluid did not affect clinical outcomes.

Evaluation of Serum S100A8/S100A9 Levels in Patients with Autoimmune Thyroid Diseases.

The correlation of S100A8/S100A9 with various inflammatory conditions, including autoimmune diseases have been reported. There is no study investigating the levels of S100A8/S100A9 in autoimmune thyroid diseases (AITD). We aimed to evaluate the level of serum S100A8/S100A9 in AITD. Case control study. Fifty patients with AITD (25 Hashimoto's thyroiditis (HT) and 25 Graves' disease (GD)) were included in the study. Twenty seven healthy subjects participated as a control group. Blood samples were obtained in the 3 months after the initiation of medical treatment. Serum levels of total antioxidant status (TAS), total oxidative status (TOS), total free sulfhydryl (SH), lipid hydroperoxide (LOOH) and S100A8/S100A9 were analyzed. The patients with AITD had significantly higher S100A8/S100A9, OSI, LOOH and TOS levels than the healthy control group. There was no significant difference between GD and HT patients in terms of S100A8/S100A9, TOS and OSI levels. S100A8/S100A9 level was positively correlated with LOOH, TOS and OSI levels but negatively correlated with -SH level in the patients with AITD. Serum S100A8/S100A9 levels were increased in patients with AITD and positively correlated with LOOH, TOS and OSI whereas negatively correlated with SH.

Alteration in mitochondrial function and glutamate metabolism affected by 2-chloroethanol in primary cultured astrocytes

The aim of this study was to explore the mechanisms that contribute to 1,2-dichloroethane (1,2-DCE) induced brain edema by focusing on alteration of mitochondrial function and glutamate metabolism in primary cultured astrocytes induced by 2-chloroethanol (2-CE), a metabolite of 1,2-DCE in vivo. The cells were exposed to different levels of 2-CE in the media for 24h. Mitochondrial function was evaluated by its membrane potential and intracellular contents of ATP, lactic acid and reactive oxygen species (ROS). Glutamate metabolism was indicated by expression of glutamine synthase (GS), glutamate–aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) at both protein and gene levels. Compared to the control group, exposure to 2-CE could cause a dose dependent damage in astrocytes, indicated by decreased cell viability and morphological changes, and supported by decreased levels of nonprotein sulfhydryl (NPSH) and inhibited activities of Na+/K+-ATPase and Ca2+-ATPase in the cells. The present study also revealed both mitochondrial function and glutamate metabolism in astrocytes were significantly disturbed by 2-CE. Of which, mitochondrial function was much vulnerable to the effects of 2-CE. In conclusion, our findings suggested that mitochondrial dysfunction and glutamate metabolism disorder could contribute to 2-CE-induced cytotoxicity in astrocytes, which might be related to 1,2-DCE-induced brain edema.

Hepatoprotective liposomal glycyrrhizin in alcoholic liver injury

IntroductionLicorice root (Glycyrrhiza glabra L.) extract and its main active component glycyrrhizin, as well as derivates are well known hepatoprotective and gastroprotective agents. Liposomal applications of these compounds may prevent low bioavailability and side-effects, e.g. pseudoaldosteronism and has further benefits, because the nanoparticles with glycyrrhizin and glycyrrhetinic acid have enhanced uptake into the liver. Our aim was to study the hepatoprotective effect of liposomal glycyrrhizin treatment during long-term alcohol consumption in rats. MethodsHarlan-Wistar (175–200g) rats were randomly allocated into three groups: control, alcoholic, ethanol and liposomal glycyrrhizin-treated. The redox status of the liver was studied with the measurement of free sulfhydryl-group concentration and hydrogen-donor ability. Transmethylation capacity was measured in dimedone adduct form. Histological examination was carried out as well. ResultsData showed mild elevation in the hydrogen-donor ability and slightly lowered free sulfhydryl level, although transmethylation capacity was significantly lowered (p<0,05) in the alcoholic group. Liposomal glycyrrhizin caused no marked histological changes. Effect of the treatment on the redox homeostasis and transmethylation capacity was beneficial. ConclusionsLiposomal glycyrrhizin treatment moderated the alcohol-related negative changes, therefore glycyrrhizin and/or its derivatives may have further impact in the treatment of alcoholic liver diseases.

Piper umbellatum L.: A medicinal plant with gastric-ulcer protective and ulcer healing effects in experimental rodent models

Ethnopharmacological relevancePiper umbellatum L. (Piperaceae) is a shrub found in the Amazon, Savannah and Atlantic Forest region of Brazil. It is widely used in folk medicine in many countries primarily for the treatment of gastric disorders. The aim of this study was to evaluate the gastroprotective and anti-ulcer effects of hydroethanolic extract of P. umbellatum (HEPu) leaves in experimental rodents. In addition, the anti-Helicobacter pylori activity of the extract was assessed. Materials and methodsThe leaves of P. umbellatum were macerated in 75% (1:3w/v) hydroethanolic solution to obtain HEPu. The gastroprotective and ulcer healing activities of HEPu were evaluated using acidified ethanol (acute) and acetic acid (chronic) gastric ulcer models in rodents. The anti-H. pylori activity was evaluated by in vitro broth microdilution assay using H. pylori cagA+ and vacA+ strain. The probable mechanism of action of HEPu was evaluated by determining gastric secretory parameters, antioxidant enzyme (catalase), non-protein sulfhydryl (glutathione) and malondialdehyde levels in gastric tissue, including pro-inflammatory (IL-1β, TNF-a, IL -17, RANTES, IFN-γ and MIP-2) and anti-inflammatory (IL-10) cytokines. ResultsHEPu demonstrated potent gastroprotection against acute ulcer induced by acidified ethanol and excellent healing effect of the chronic ulcer induced by acetic acid. The gastroprotective activity in acidified ethanol is partly attributed to the antioxidant mechanisms, while anti-secretory, anti-inflammatory and regeneration of the gastric mucosa are evoked as part of its antiulcer mechanism of action. The gastric ulcer healing of HEPu also involves restoration of the altered cytokines levels to near normal. However, it has no in vitro anti-H. pylori activity. ConclusionThe results of this study showed that HEPu possesses preventive and curative effects in experimental models of gastric ulcers in animals. These effects are partially dependent on antioxidant, antisecretory, anti-inflammatory and mucosa regeneration. It is independent of anti-H. pylori activity, with substances probably responsible for the pharmacological activity being flavonoids, quercetin and rutin. These results support the popular use of P. umbellatum leaves in the treatment of peptic ulcers.

Influence of different frequencies of deep water running on oxidative profile and insulin resistance in obese women

IntroductionObesity is associated with health damages related to increased oxidative stress and insulin resistance. Aerobic exercise can be an ally in reducing the prevalence of obesity and its consequences. This study evaluated the effect of deep water running in two frequencies, with the same intensity, in anthropometric parameters, oxidative profile and insulin resistance in obese and sedentary women. MethodsThe study included 24 women with ages ranging from 47.33 ± 2.98 years old and body mass index of 33.39 ± 0.77 kg/m2. They were divided into two groups: aerobic training 1 (AT1) (5 days/week) and aerobic training 2 (AT2) (3 day/week), both with moderate intensity (50–75% VO2max – Borg Scale), for 60 min each session. Anthropometric measurements, oxidative stress and insulin resistance were evaluated before and after the 26 training sessions. ResultsAT1 training provoke a significant reduction in anthropometric parameters, lipids peroxidation (TBARS) and protein oxidation (carbonyl), and increased enzymatic activity of superoxide dismutase and catalase (p < 0.05). On the other hand, AT2 reduced waist circumference, sulfhydryl levels and GPx activity; however, this training did not alter insulin resistance parameters. ConclusionsThe data suggest that the deep water running performed 5 days per week (AT1) proved to be more effective in reducing obesity rates. For this reason, this training could be an important choice to help reduce the anthropometric parameters and oxidative damage, and increase antioxidant defenses in obese women.