Reduced GSH Content Research Articles

Role of thymoquinone and ebselen in the prevention of sodium arsenite-induced nephrotoxicity in female rats.

The aim of this study is to investigate the protective effects of thymoquinone (TQ) and ebselen (Eb) on arsenic (As)-induced renal toxicity in female rats. Sodium arsenite was orally administrated at a dose of 20 mg/kg body weight daily for 28 days, either alone or 1 h before TQ (10 mg/kg) or Eb (5 mg/kg) administration. Renal tissue As concentration and oxidative stress markers, including lipid peroxidation (LPO), nitrite/nitrate, and glutathione (GSH) levels, were determined. In addition to the oxidative stress response, antioxidant enzyme activities including that of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were measured. Exposure to As elicited a significant increase in As concentration and significant modifications to the redox state of the kidney, as was evidenced by a significant elevation in LPO and nitrite/nitrate concentration, with a concomitant reduction in GSH content and antioxidant enzyme activity. The oxidant/antioxidant imbalance observed in As toxicity was associated with a significant elevation in renal tumor necrosis factor α, interleukin 6, B-cell lymphoma 2 (Bcl-2)-associated X protein, and caspase 3 levels, in addition to a significant decrease in Bcl-2 levels. Post-administration of TQ and Eb markedly prevented As-induced oxidative stress, inflammation, apoptosis, and As accumulation in the renal tissue and reduced histological renal damage. These findings demonstrate that TQ, the main bioactive phytochemical constituent of Nigella sativa seed oil, and Eb, an organoselenium compound, could significantly inhibit As-induced oxidative damage, apoptosis, and inflammation, and significantly attenuate the accumulation of As in renal tissues by facilitating As biomethylation and excretion.

Effects of Propolis on Oxidative Stress in Rabbits Undergoing Implant Surgery

Abstract Objectives: Implants are the natural or artificial materials that replace a missing tissue or organ. When that missing part is one or more teeth, there is a need for the reconstructive material that supports prosthetic restoration, which is called dental implant. Many local and general factors affect bone healing. Recent studies show that, free radicals are among the factors that affect the bone healing. This study is planned presuming propolis could have positive effects on oxidative stress and bone healing after implantation due to its antioxidant nature, its effects on accelerating the bone formation, wound healing and increasing the bone density. Materials and Methods: 24 New Zealand white rabbits were used in the study. Animals were divided into 3 groups, each consisting of 8 rabbits. Implants were placed into the proximal region of tibia unilaterally. Propolis was applied to slots before placing the implants on local group and systemically applied after implantation on systemic group. Results: SOD activity was increased in both propolis groups compared to control group, but it was not statistically significant. Catalase level of the systemic group was found to be significantly higher. MDA level in serum, which indicates lipid peroxidation, was significantly lower in both propolis groups compared to control group. Reduced GSH concentration in granulation tissue was increased in rabbit groups treated with propolis compared to control group. There were no significant differences in calcium and phosphorus values, however it can be said that there is a significant increase in vitamin D amount of local and systemic groups. Conclusion: Propolis has antioxidant, antimicrobial and many other biological activities. Due to the features that it has, propolis has a positive effect on osseointegration and bone healing. The comparison of systemic and local group showed that, systemically given propolis provided better results than local application.

Vitamin E alleviates phoxim-induced toxic effects on intestinal oxidative stress, barrier function, and morphological changes in rats.

Phoxim is an organic phosphorus pesticide that remains easily in the environment, such as human food and animal feed. The objective of this study was to explore the effect of vitamin E on phoxim-induced oxidative stress in the intestinal tissues of Sprague-Dawley (SD) rats. Forty-eight Sprague-Dawley rats were randomly assigned to a control group and three treatment groups: treatment group 1 (phoxim: 20mg/kg·BW), treatment group 2 (phoxim: 180mg/kg·BW), and treatment 3 (vitamin E + phoxim: 200mg/kg·BW + 180mg/kg·BW). Phoxim was given by gavage administration once a day for 28days. The results showed that phoxim significantly reduced jejunum villus height in rats (P< 0.05), and decreased the mRNA expression of junction protein genes of rats, including Occlidin and Claudin-4 (P < 0.05). Phoxim reduced GSH content and T-AOC level in the intestinal mucosa (P < 0.05). The mRNA expression levels of oxidative stress-related genes (Nrf2 and GPx2) were decreased. The mRNA expression of SOD was significantly increased. In addition, phoxim increased the level of interleukin-6 (IL-6) in jejunum mucosa and significantly reduced the level of IL-8 in ileum mucosas, while significantly increased TNF-α secretion. The mRNA expression levels of IL-1β, IL-6, and IL-8 were significantly decreased, and mRNA expression of TNF-α was significantly increased (P < 0.05). Phoxim also increased the DNA expression of total cecal bacteria and Escherichia coli, inhibited the DNA expression of Lactobacillus and destroyed the intestinal barrier. Two hundred milligrams per kilogram BW vitamin E reduced the effect of phoxim on intestinal structure, alleviated the oxidative stress in intestinal tissue, and decreased the level of TNF-α. The mRNA expressions of antioxidative stress genes (SOD and GPx2) were significantly increased. The DNA expression level of Lactobacillus was significantly increased. In conclusion, vitamin E helped reduce the toxicity of organophosphate pesticides, such as phoxim on rat intestinal tissue.

Evaluation of oxidative stress indices after exposure to malathion and protective effects of ascorbic acid in ovarian tissue of adult female rats.

BackgroundMalathion is one of organophosphate pesticides that is extensively used in farming and crops to control pests. Malathion induces oxidative stress in the various tissues such as the reproductive system.ObjectiveTo determine the effects of malathion on malondialdehyde (MDA) level and glutathione (GSH) content in female rat ovary tissue as well as to assess the protective role of Ascorbic Acid.MethodsThis study was carried out at the Department of Anatomy and Cell Biology (School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran) in 2015. In this experimental study, 30 adult, female, Wistar rats (weight range: 200–250 g) were divided into five groups, each group consisting of six rats: control group (no interventions), sham group (normal saline 0.9% 50 mg/kg), experimental group 1 (Ascorbic Acid 200 mg/kg), experimental group 2 (malathion 50 mg/kg), and experimental group 3 (malathion 50 mg/kg + Ascorbic Acid 200 mg/kg). Malathion, solvents and Ascorbic Acid were injected intraperitoneally. After two weeks, the animals were anaesthetized with intraperitoneal ketamine/xylazine (60 and 6 mg/kg, respectively) and then scarified, and the right ovarian was used to measure levels of MDA, a marker of lipid peroxidation, and GSH content. Data were analyzed by SPSS version 16, using descriptive statistics, One Way ANOVA, and Tukey-Kramer test. A p-value <0.05 was set as significance level.ResultsThis study has shown that malathion increased MDA level and reduced GSH content compared with the control group (p<0.001). Also, administration of malathion in combination with Ascorbic Acid, reduced MDA level and increased the GSH content in rat ovarian tissue.ConclusionMalathion induced lipid peroxidation and Oxidative stress in the ovarian of Rats. In addition, it appears that Ascorbic Acid, due to its antioxidant, can recover malathion-induced poisonous changes.

Expression of thioredoxin-2 in human lens epithelial cells with oxidative damage and its significance

To explore whether thioredoin-2 (Trx-2) is involved in the development of cataract and to study the effect of Trx-2 on hydrogen peroxide (H2O2)-induced injury in human lens epithelial cells. Methods: A total of 10 volunteers (removing the lens due totraumatism) and 30 patients received phacoemulsification (age more than 60 years) were selected. The expression of Trx-2 protein in lens epithelial cells from cataract patients and volunteers were detected by the immunohistochemical streptavidin-peroxidase (SP) method. SRA01/04 cells were cultured and were divided into six groups according to different treatment: a control group, H2O2-treated groups at 20, 50 or 100 μmol/L, a negative control group (transfected with pCMV6 plasmid plus 100 μmol/L H2O2), and a Trx-2 overexpression group (transfected with pCMV6-Trx-2 plasmid plus 100 μmol/L H2O2). Methyl thiazolyltetrazolium (MTT) assay and flow cytometry was performed to measure the cell viability and apoptosis for SRA01/04 cells, respectively. The activities of superoxide dismutase (SOD) and catalase (CAT), the content of glutathione (GSH) and malondialdehyde (MDA) in human lens epithelial cells were measured via chemical chromatometry. Western blot was used to measure the protein levels of Trx-2, B-cell lymphoma 2 protein (Bcl-2), Bcl-2 associated X protein (Bax) and caspase-3. Results: Compared with the volunteers, the expression of Trx-2 was significantly decreased in lens epithelial cells in patients with cataract (P<0.05). Compared with the control group, the expression of Trx-2 protein in the 20, 50 or 100 μmol/L H2O2 groups was decreased (all P<0.05). Compared with the control group, the cell survival rates were decreased in the 100 μmol/L H2O2 group and the negative control group (both P<0.05), along with enhanced apoptotic rates, inhibited cellular SOD activities and CAT activities, reduced GSH contents, augmented MDA contents, down-regulated Trx-2 and Bcl-2 expression and up-regulated Bax and caspase-3 expression (all P<0.05). Compared with the negative control group, the cell survival rate was increased in the Trx-2 overexpression group (P<0.05), along with suppressed apoptosis, increased SOD activities and CAT activities, elevated GSH contents, decreased MDA content, up-regulated Trx-2 and Bcl-2 expression and down-regulated Bax and caspase-3 expression (P<0.05). Conclusion: Trx-2 might be involved in the apoptosis of lens epithelial cells in patients with cataract. The overexpression of Trx-2 obviously attenuated H2O2-induced injury of human lens epithelial cells, which might be associated with the inhibition of H2O2-mediated oxidative stress.

Hydrogen sulphide increase the tolerance to alkalinity stress in cabbage plants ( Brassica oleracea L. 'Bronco')

The present study investigates how the application of hydrogen sulphide (0.5 mM of NaHS) in Brassica oleracea L. ‘Bronco’ influences the processes involved in glutathione homeostasis and tolerance to alkaline stress (50 mM NaHCO3:Na2CO3). According to our results, alkaline stress increases the O2− content, lipid peroxidation, and the activities of the enzymes glyoxalase I (Gly I) and glyoxalase II (Gly II) that detoxify methylglyoxal (MG) while decreasing biomass, the activity of superoxide dismutase (SOD), the activity of enzymes involved in glutathione (GSH) synthesis and in the AsA-GSH cycle, as well as the content in reduced glutathione and the different forms of ascorbate (AsA). On the other hand, the application of NaHS improved the antioxidant response, inducing SOD activity and improving processes involved in glutathione homeostasis, boosting the reduced GSH content as well as the activity of key enzymes in glutathione synthesis and in the AsA-GSH cycle. Consequently, the application of H2S in the form of NaHS at a concentration of 0.5 mM could fortify the tolerance of Brassica oleracea L. ‘Bronco’ against alkaline stress.

The Problem of Glutathione Determination: a Comparative Study on the Measurement of Glutathione from Plant Cells

The level and redox status of glutathione is a good indicator for the rate of oxidative stress and eco-toxicological injury in plant cells and subcellular organelles. Thus the determination of GSH and its redox status has special importance. A variety of spectrophotometric and HPLC methods are available to measure glutathione (GSH). The spectrophotometric DTNB-GSH recycling assay is specific due to the application of glutathione reductase, it is rather quick and easy to perform, not surprising that it is rather popular. In the present study we make an attempt to compare the DTNB-GSH recycling assay and the more sophisticated, but difficult monochlorobimane (mBCl)-HPLC method to choose the one that best suits for eco-toxicological and plant stress investigations. We found that the acidification by sulphosalicylic acid (SSA) used for the stabilization of samples for DTNB-GSH recycling assay gives lower efficiency to this method than the formation of mBCl-GSH fluorescent conjugate. The measurable GSH contents were lower in the case of DTNB-GSH recycling assay than in the case of GSH-mBCl conjugates determined by HPLC with fluorescence detection. The auto-oxidation could almost perfectly be prevented by the presence of mBCl in the organelle isolation buffer. Furthermore, this way the reduced GSH content of organelles could be determined much more precisely. However, it is worth to note that the application of mBCl significantly elevates the cost of GSH determination, especially in case of cell organelles.

Enhanced Redox State and Efficiency of Glucose Oxidation With miR Based Suppression of Maladaptive NADPH-Dependent Malic Enzyme 1 Expression in Hypertrophied Hearts.

Metabolic remodeling in hypertrophic hearts includes inefficient glucose oxidation via increased anaplerosis fueled by pyruvate carboxylation. Pyruvate carboxylation to malate through elevated ME1 (malic enzyme 1) consumes NADPH necessary for reduction of glutathione and maintenance of intracellular redox state. To elucidate upregulated ME1 as a potential maladaptive mechanism for inefficient glucose oxidation and compromised redox state in hypertrophied hearts. ME1 expression was selectively inhibited, in vivo, via non-native miR-ME1 (miRNA specific to ME1) in pressure-overloaded rat hearts. Rats subjected to transverse aortic constriction (TAC) or Sham surgery received either miR-ME1 or PBS. Effects of ME1 suppression on anaplerosis and reduced glutathione (GSH) content were studied in isolated hearts supplied 13C-enriched substrate: palmitate, glucose, and lactate. Human myocardium collected from failing and nonfailing hearts during surgery enabled RT-qPCR confirmation of elevated ME1 gene expression in clinical heart failure versus nonfailing human hearts (P<0.04). TAC induced elevated ME1 content, but ME1 was lowered in hearts infused with miR-ME1 versus PBS. Although Sham miR-ME1 hearts showed no further reduction of inherently low anaplerosis in normal heart, miR-ME1 reduced anaplerosis in TAC to baseline: TAC miR-ME1=0.034±0.004; TAC PBS=0.081±0.005 (P<0.01). Countering elevated anaplerosis in TAC shifted pyruvate toward oxidation in the tricarboxylic acid cycle. Importantly, via the link to NADPH consumption by pyruvate carboxylation, ME1 suppression in TAC restored GSH content, reduced lactate production, and ultimately improved contractility. A maladaptive increase in anaplerosis via ME1 in TAC is associated with reduced GSH content. Suppressing increased ME1 expression in hypertrophied rat hearts, which is also elevated in failing human hearts, reduced pyruvate carboxylation thereby normalizing anaplerosis, restoring GSH content, and reducing lactate accumulation. Reducing ME1 induced favorable metabolic shifts for carbohydrate oxidation, improving intracellular redox state and enhanced cardiac performance in pathological hypertrophy.

Thiocyclam-Induced Reproductive Toxicity, Oxidative Stress and Genomic DNA Damage in Testicular Tissues of Rats: Protective Effects of Green Tea Extract

Thiocyclam is a broad-spectrum nereistoxin analogue insecticide which widely used for agricultural applications. The aim of this study is to investigate the protective effects of green tea (as a principal source of antioxidants) on Thiocyclam-Induced Reproductive Toxicity, Oxidative Stress and Genotoxicity in adult Male Rats. Forty adult male rats were randomly assigned into four groups: control, Thiocyclam (ThC, 15.98 mg/kg b.w.), green tea extract (GTE, 2% w/v as the sole beverage) and (ThC+GTE) group. After 65 days of treatment, blood samples and testicular tissue were collected for measuring the oxidative stress parameters, testosterone level and DNA damage, whereas the reproductive organs were weighed, and Semen analysis and testicular histopathological studies was done. The results revealed that Thiocyclam administration induce a significant reduction in sperm count, sperm motility as well as testosterone level, while increased sperm abnormality. In addition to increased testicular tissue Malondialdehyde (MDA), reduced GSH content and testicular DNA damage evidenced by comet assay. The histological examination of testes revealed degenerative change and disorganization of seminiferous tubules with incomplete spermatogenesis. On the contrary, GTE played a potential protective effect against Thiocyclam induced oxidative stress as well as alleviate the reproductive toxicity by improving the oxidative status, decrease oxidative DNA damage, improving semen characteristics and protective effect against testicular damage, highlighting the protective and therapeutic potentiality of green tea against pesticide-induced reproductive toxicity and its health benefits.

Crosstalk between Nrf2 and YAP contributes to maintaining the antioxidant potential and chemoresistance in bladder cancer.

Redox adaptation plays an important role in cancer cells drug resistance. The antioxidant response is principally mediated by the transcription factor Nrf2, that induces the transcriptional activation of several genes involved in GSH synthesis, chemoresistance, and cytoprotection. YAP is emerging as a key mediator of chemoresistance in a variety of cancers, but its role in controlling the antioxidant status of the cells is yet elusive. Here, we show that impairing YAP protein expression reduced GSH content and Nrf2 protein and mRNA expression in bladder cancer cells. Moreover, inYAP knocked down cells the expression of FOXM1, a transcription factor involved in Nrf2 transcription, was down-regulated and the silencing of FOXM1 reduced Nrf2 expression. On the other hand, the silencing of Nrf2, as well as the depletion of GSH by BSO treatment, inhibited YAP expression, suggesting that cross-talk exists between YAP and Nrf2 proteins. Importantly, we found that silencing either YAP or Nrf2 enhanced sensitivity of bladder cancer cells to cytotoxic agents and reduced their migration. Furthermore, the inhibition of both YAP and Nrf2 expressions significantly increased cytotoxic drug sensitivity and synergistically reduced the migration of chemoresistant bladder cancer cells. These findings provide a rationale for targeting these transcriptional regulators in patients with chemoresistant bladder cancer, expressing high YAP and bearing a proficient antioxidant system.

Abstract B22: Selective responses of HepG2 cancer cells and primary hepatocytes to growth regulatory effects of Fe (II) and herbal tea flavonoids

Abstract The role of flavonoids as chemopreventive agents is complex due to flavonoid-cell interactions affecting oxidative stress parameters known to play a key role in cell growth and survival. The current study investigated differential anti-proliferative and pro-apoptotic effects and modulation of oxidative status by unfermented (Rg) and polyphenol-enriched (PERg) rooibos (Aspalathus linearis) extracts, while the interaction with Fe (II) in liver cancer (HepG2) cells and primary rat hepatocytes were critically assessed. In contrast to the rooibos extracts, Fe (II) increased cell proliferation in HepG2 cells while in primary hepatocytes both viability and proliferation was decreased. Fe (II) countered the reduction in cell viability and inhibition of proliferation by either Rg or the PERg extracts in HepG2 cells, implying a protective effect. Similar treatments did not significantly alter the growth parameters in primary hepatocytes. Pre-exposure of HepG2 cells to Fe (II) tended to also counteract the reduction in cell viability induced by Rg and PERg while no effect was noticed in primary rat hepatocytes. The PERg extract was more prominent in affecting apoptosis in HepG2 cells, while the opposite was noticed in primary hepatocytes where Rg was more effective. In both cases, Fe (II) counteracted the induction of apoptosis, which was associated with an increase in cell viability. Pre-exposure to iron provide little protective effects, although the induction of apoptosis was markedly reduced, suggesting that a memory effect existed following Fe (II) exposure in both cell culture systems. Thus, Fe (II) decreased the susceptibility of the cells to the effects of the rooibos extracts presumably due to the chelating properties by counteracting the pro-oxidant effects of the rooibos flavonoids. The modulation of cell viability and proliferation by rooibos flavonoids are related to their redox-sensitivity. Both extracts significantly increased glutathione (GSH) levels in HepG2 cells, while only the highest Rg extract concentration resulted in an increase in primary rat hepatocytes. This response is indicative of an antioxidant cellular response presumably by adapting to an oxidative insult effect by the rooibos extracts. Co-exposure with Fe (II) in HepG2 cells counteracted the effects of the rooibos extracts by normalizing the GSH content similar to that of the control level. Pre-exposing cells to Fe (II) showed little memory effect on the reduction of GSH content as the levels was significantly increased above that affected by the rooibos extracts in HepG2 cells suggesting an increased insult on the redox status. In primary hepatocytes a similar effect was noticed although to a far lesser extent. The activity of glutathione peroxidase (GPx) was significantly decreased by the rooibos extracts, while Fe (II) tended to counteract the reduction during the combined and pre-exposure models in both cell culture systems. The reduction in the GPx activity was more prominent in the primary hepatocytes. Understanding the underlying flavonoid/Fe (II) interactions and the modulation of cellular oxidative status and cell growth parameters will provide valuable information to reduce the risk of exposure to polyphenol-enriched neutraceuticals and food supplements in humans. Citation Format: Sedicka Samodien, Maryna de Kock, Sonja Swanevelder, Elizabeth Joubert, Wentzel Gelderblom. Selective responses of HepG2 cancer cells and primary hepatocytes to growth regulatory effects of Fe (II) and herbal tea flavonoids [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B22.

Protective effects of gallic acid against methotrexate-induced toxicity in rats

Background: Methotrexate, as a chemotherapy drug, can cause chronic liver damage and oxidative stress. Aim of this study was to evaluate the preventive effect of gallic acid (GA) on methotrexate (MTX)-induced oxidative stress in rat liver.Methods: Twenty-eight male rats were randomly divided into four groups as control, MTX (20 mg/kg, i.p.), MTX + GA (30 mg/kg/day, orally) and GA treated. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were used as biochemical markers of MTX-induced hepatic injury. Malondialdehyde (MDA) and glutathione (GSH) levels and hepatic antioxidant enzymes activities including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assayed in liver tissue. The expression of SOD2 and GPx1 genes were evaluated by real-time RT-PCR and liver histopathology was evaluated by light microscopy.Results: The result obtained from current study showed that GA remarkably reduced MTX-induced elevation of AST, ALT and ALP and increased MTX-induced reduction in GSH content, GPx, CAT and SOD activity as well as GPx1 and SOD2 gene expressions. Histological results showed that MTX led to liver damage and GA could improve histological changes.Conclusions: Our results indicate that GA ameliorates biochemical and oxidative stress parameters in the liver of rats exposed to MTX.

Phytochemical Investigation, Antitumor Activity, and Hepatoprotective Effects of Acrocarpus fraxinifolius Leaf Extract.

Preclinical Research Nine known phenolic compounds were isolated from an aqueous methanolic extract of Acrocarpus fraxinifolius Weight and Arn leaves (AFL) family Fabaceae. This extract of AFL contained approximately 169 mg gallic acid/g as assessed by HPLC. The AFL extract had marginal antitumor activity (IC50 > 200 µL/mL) but showed a concentration-dependent hepatoprotective effect against CCl4 -induced hepatotoxicity in vitro. Cell viability was increased, ALT and AST activity declined and reduced GSH concentration and SOD activity were restored as compared with silymarin. In vivo concurrent administration of AFL extract (500 mg/kg po) showed a hepatoprotective effect against gamma irradiation and CCl4 as evidenced by reduction of TNF-α, interleukin-6, malondialdehyde, nitric oxide, DNA fragmentation, caspase-3 activity, and downregulation of its m-RNA level and decreased proapoptotic protein Bax expression. AFL extract enhanced glutathione peroxidase, superoxide dismutase, and catalase activities, reduced glutathione concentrations and upregulated the expression of antiapoptotic Bcl-2. The extract could ameliorate hepatic injuries induced by gamma irradiation and CCl4 in rats suggesting potent hepatoprotective activity. Drug Dev Res 78 : 210-226, 2017. © 2017 Wiley Periodicals, Inc.

Impact of radiofrequency radiation on DNA damage and antioxidants in peripheral blood lymphocytes of humans residing in the vicinity of mobile phone base stations

ABSTRACTRadiofrequency radiations (RFRs) emitted by mobile phone base stations have raised concerns on its adverse impact on humans residing in the vicinity of mobile phone base stations. Therefore, the present study was envisaged to evaluate the effect of RFR on the DNA damage and antioxidant status in cultured human peripheral blood lymphocytes (HPBLs) of individuals residing in the vicinity of mobile phone base stations and comparing it with healthy controls. The study groups matched for various demographic data including age, gender, dietary pattern, smoking habit, alcohol consumption, duration of mobile phone use and average daily mobile phone use. The RF power density of the exposed individuals was significantly higher (p < 0.0001) when compared to the control group. The HPBLs were cultured and the DNA damage was assessed by cytokinesis blocked micronucleus (MN) assay in the binucleate lymphocytes. The analyses of data from the exposed group (n = 40), residing within a perimeter of 80 m of mobile base stations, showed significantly (p < 0.0001) higher frequency of micronuclei when compared to the control group, residing 300 m away from the mobile base station/s. The analysis of various antioxidants in the plasma of exposed individuals revealed a significant attrition in glutathione (GSH) concentration (p < 0.01), activities of catalase (CAT) (p < 0.001) and superoxide dismutase (SOD) (p < 0.001) and rise in lipid peroxidation (LOO) when compared to controls. Multiple linear regression analyses revealed a significant association among reduced GSH concentration (p < 0.05), CAT (p < 0.001) and SOD (p < 0.001) activities and elevated MN frequency (p < 0.001) and LOO (p < 0.001) with increasing RF power density.

A traditional poly herbal medicine \u201cLe Pana Guliya\u201d induces apoptosis in HepG2 and HeLa cells but not in CC1 cells: an in vitro assessment

“Le Pana Guliya” (LPG) is a polyherbal formulation which is used to treat different types of cancers in traditional medicine. In this study we describe in vitro efficacy and mechanism of action of LPG on two cancer cell lines (HepG2 and HeLa) compared with a normal cell line CC1. The MTT, LDH assays and protein synthesis were used to study antiproliferative activity of LPG while NO synthesis and GSH content were assayed to determine the oxidative stress exerted by LPG. Rhodamine 123 staining, caspase 3 activity, DNA fragmentation and microscopic examination of cells stained with ethidium bromide/acridine orange were used to identify the apoptosis mechanisms associated with LPG. The LPG showed the most potent antiproliferative effect against the proliferation of HepG2 and HeLa cells with an EC50 value of 2.72 ± 1.36 and 19.03 ± 2.63 µg/mL for MTT assay after 24 h treatment respectively. In contrast, CC1 cells showed an EC50 value of 213.07 ± 7.71 µg/mL. Similar results were observed for LDH release. A dose dependent decrease in protein synthesis was shown in both cancer cell types compared to CC1 cells. The reduction of GSH content and elevation of cell survival with exogenous GSH prove that the LPG act via induction of oxidative stress. LPG also stimulates the production of NO and mediates oxidative stress. Rhodamine 123 assay shows the mitochondrial involvement in cell death by depletion of Δψ inducing downstream events in apoptosis. This results in increase in caspase-3 activity eventually DNA fragmentation and LPG induced apoptotic cell death. In conclusion the present study suggested that the LPG exerted an anticancer activity via oxidative stress dependent apoptosis. Therefore present study provides the scientific proof of the traditional knowledge in using LPG as an anticancer agent.

Oxidative injury caused by individual and combined exposure of neonicotinoid, organophosphate and herbicide in zebrafish.

The greatest challenge in environmental toxicology is to understand the effects of mixture toxicity as environmental pollutants co-exist and exhibit combined effects. Thus, it is necessary to evaluate the mixture toxicity associated with two or more co-existing compounds. Pesticides are widely used to control pest, they are ubiquitous in nature and present in all environmental components. Pesticide residue can be detected in almost all components of environment and food samples. Imidacloprid (IMD) (neonicotinoid), dichlorvos (DIC) (organophosphate) and atrazine (ATZ) are three widely used pesticides for commercial uses. Present work includes the assessment of effects of individual exposure of IMD (27.5mg/L), DIC (15mg/L), and ATZ (03mg/L) and in combination of three (CMD) (13.75+7.5+1.5mg/L IMD, DIC & ATZ, respectively) in terms of LPO, GSH content and antioxidant enzymes activities (superoxide dismutase, catalase and glutathione peroxidase) in zebrafish (Danio rerio), exposed for 24h. CMD group exhibits highest lipid peroxidation than other individually exposed groups. Similarly, the activities of antioxidant enzymes were highest in CMD group with reduced GSH content. Results indicate that exposure to mixture of pesticides develops synergistic effects which were more toxic in compare to individual exposure and also produce toxicity in all examined tissues rather than selective organ toxicity.

Hepatoprotective activities of Lappula barbata and Plantago holosteum against paracetamol induced liver damage in rats and their in vitro antioxidant effects

Plantago and Lappula species are represented by 24 and 7 species in Turkey, respectively [1]. Plantago species are used for liver disease in folk medicine and contain aucubin and flavonoids [2]. Lappula species showed antiviral and anti-inflammatory activities. Boraginaceous plants are characterized by high levels of polyunsaturated fatty acids [3]. In order to evaluate medicinal uses of Plantago holosteum Scop. (PH) and Lappula barbata Bieb. (LB), the potential hepatoprotective effect of the ethanol (70%) extracts [PH: 4.38 g (yield: 17.0%); LB: 2.75 g (yield: 10.9%)] of LB and PH in a rat model of PCM induced liver damage were investigated. PCM administration caused severe hepatic damage in rats [4] as evidenced by elevated serum activities of ALT, AST, ALP, ?-GT and serum level of BRN while decreased serum levels of TP and ALB. It elevated MDA, decreased GSH levels, GPx, SOD and CAT activities in liver homogenates. Administration of LB (400 mg/kg) and PH (200 and 400 mg/kg) extracts for 7 days before PCM inhibited the elevation of the serum activities of ALT, AST, ALP and ?-GT enzymes and serum level of BRN. They elevated the serum level of TP (27.45, 29.41 and 33.33%, respectively). PCM-induced lipid peroxidation was reduced following administration of LB (25.21%) and PH (22.84 and 28.55%, respectively) at same doses. They increased the activities of the antioxidant enzymes (GPx, SOD, CAT) in liver homogenates and reduced GSH concentration (30.76, 30.06 and 36.36%, respectively) at same doses. The results revealed evident antioxidant activities for both extracts. The histopathological analysis suggested that both extracts obviously alleviated the degree of liver damage due to PCM administration. They protected against focal necrosis of hepatocytes and hepatocytes apoptosis induced by PCM. The present study suggests that LB and PH possess hepatoprotective activities that could be partly attributed to their antioxidant effects and the aucubin content of PH.

Role of Carvedilol in Prevention of Adrenaline Induced Myocardial Infraction in Experimental Animal

Introduction: Acute myocardial infarction (AMI) is the most important form of ischemic heart disease (IHD). Coronary artery disease (CAD) is an increasingly important medical and public health problem and is the leading cause of mortality in Bangladesh. AMI is the rapid development of myocardial necrosis caused by a critical imbalance between the oxygen supply and demand of the myocardium. Total occlusion of the coronary arteries for more than 4-6 hrs results in irreversible myocardial necrosis, but reperfusion within this period can salvage the myocardium and reduce morbidity and mortality.&#x0D; Objectives: To assess the role of carvedilol in prevention of adrenaline induced cardiac damage in experimental animal.&#x0D; Materials and Methods: This experimental study was carried out in the department of pharmacology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka for a period of One year spanning from July 2004 to June 2005. Fifty two healthy rats of Long Evan Norwegian strains, 3-4 months of ages of both sexes, weight between 180-220g were used. The experiment was divided into two parts: Part-I and Part-II. Thirty two rats were selected for Part-I experiment and subdivided into Group-I and Group-II. In Part-II experiment, 20 rats were selected and placed as Group-III. Group-I (12 rats) of control group was treated with 02 doses of inj distilled water (D/W) subcutaneously (S.C.) 24 hrs apart and serum creatine kinase-MB (CK-MB) level and hepatic and cardiac reduced glutathione (GSH) contents were estimated from 06 (Group-Ia) rats after 12 hrs and serum aspartate aminotransferase (AST) level and hepatic and cardiac reduced GSH contents were estimated from 06 (Group-Ib) rats after 24 hrs of 2nd inj of D/W. Group-II (20 rats) was treated with 02 doses of inj adrenaline (2mg/kg) S.C. in 24 hrs interval and in above mentioned way serum CK-MB level, GSH (hepatic and cardiac) contents and serum AST and GSH (hepatic and cardiac) contents were estimated 12 hrs and 24 hrs after the 2nd inj of adrenaline respectively. In experimental group (Group-III) all the rats (20) were treated with carvedilol (1 mg/kg) orally for 14 consecutive days and then were given 02 doses of inj adrenaline with the interval of 24 hrs and again serum CK-MB level and GSH (hepatic and cardiac) contents were estimated from half of the rats (10) after 12 hrs of injection and serum AST level and GSH (hepatic and cardiac) contents were measured from half of the rats (10) after 24 hrs of 2nd injection of adrenaline.&#x0D; Results: Adrenaline (2mg/kg) induced myocardial damage was evaluated biochemically by significant (P˂0.001) increase in CK-MB and AST levels. Free radical production following adrenaline induced myocardial damage was reflected by significant (P˂0.001) depletion in hepatic and cardiac reduced glutathione (GSH) contents. Cardioprotection provided by carvedilol pretreatment in adrenaline induced myocardial infarction was assessed by significant prevention of increase in serum CK-MB and AST levels. Antioxidant property of carvedilol was evaluated by significant (P&lt;0.001) prevention of depletion in hepatic and cardiac GSH contents. The results of the study indicated that carvedilol pretreatment provided effective prevention in adrenaline induced myocardial damage and also provided effective antioxidative action.&#x0D; Conclusion: This study indicated that adrenaline administration induced myocardial damage as evidenced by increase in serum CK-MB and AST levels which was associated with free radical production as reflected by depletion in hepatic and cardiac GSH contents. It was observed that carvedilol through their antioxidant property in addition to their β-blocking effect prevents free radical mediated injury of catecholamine assault following MI.&#x0D; Journal of Armed Forces Medical College Bangladesh Vol.12(2) 2016: 127-134

Antioxidant and Protective Effects of Angipars™, a New Herbal Drug, on Hydrogen Peroxide-Induced Oxidative Stress in HepG2 and A549 Cell Lines

: In the present study, free-radical-scavenging and antioxidant activities of Angipars™, an herbal extract which has been registered as a therapeutic drug for diabetic foot ulcer, were examined. We investigated the protective effect of Angipars™ on cytotoxicity produced by exposure to H2O2 (300 µM) in HepG2 and A549 cell lines, using MTT assay, glutathione (GSH) content analysis, and lipid peroxidation evaluation. Angipars™ concentrations of 1 - 20 µg/mL effectively scavenged DPPH radicals in a dose-dependent manner, while hydroxyl-radical-scavenging activity was only observed at high doses. Following a 4-hour cell treatment with H2O2, a noticeable decline in cell viability was observed, which was alleviated by Angipars™ (pre-incubated for 24 hours) at high doses. Angipars™ significantly reduced GSH content and products of lipid peroxidation, resulting from H2O2 exposure. These findings suggest that Angipars™ exhibits positive antioxidant activity in vitro and can attenuate H2O2-induced cytotoxicity; in fact, this may represent the cellular mechanisms for its cytoprotective activity.

Possible involvements of glutamate and adrenergic receptors on acute toxicity of methylphenidate in isolated hippocampus and cerebral cortex of adult rats.

Neurodegeneration induced by methylphenidate (MPH), as a central stimulant with unknown long-term consequences, in adult rats' brain and the possible mechanisms involved were studied. Rats were acutely treated with MPH in the presence and absence of some receptor antagonists such as ketamine, topiramate, yohimbine, and haloperidol. Motor activity and anxiety level in rats were monitored. Antioxidant and inflammatory parameters were also measured in isolated hippocampus and cerebral cortex. MPH-treated groups (10 and 20 mg/kg) demonstrated anxiety-like behavior and increased motor activity. MPH significantly increased lipid peroxidation, GSSG content, IL-1β and TNF-α levels in isolated tissues, and also significantly reduced GSH content, superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) activities in hippocampus and cerebral cortex. Pretreatment of animals by receptor antagonists caused inhibition of MPH-induced motor activity disturbances and anxiety-like behavior. Pretreatment of animals by ketamine, topiramate, and yohimbine inhibited the MPH-induced oxidative stress and inflammation; it significantly decreased lipid peroxidation, GSSG level, IL-1β and TNF-α levels and increased GSH content, SOD, GPx, and GR activities in hippocampus and cerebral cortex of acutely MPH-treated rats. Pretreatment with haloperidol did not cause any change in MPH-induced oxidative stress and inflammation. In conclusion, acute administration of high doses of MPH can cause oxidative and inflammatory changes in brain cells and induce neurodegeneration in hippocampus and cerebral cortex of adult rats and these changes might probably be mediated by glutamate (NMDA or AMPA) and/or α2 -adrenergic receptors.