Arsenic-induced Oxidative Damage Research Articles

Melatonin attenuates liver injury in arsenic-treated rats: The potential role of the Nrf2/HO-1, apoptosis, and miR-34a/Sirt1/autophagy pathways.

Arsenic is a toxic metalloid found in the environment in different organic and inorganic forms. Molecular mechanisms implicated in arsenic hepatotoxicity are complex but include oxidative stress, apoptosis, and autophagy. The current study focused on the potential protective capacity of melatonin against arsenic-induced hepatotoxicity. Thirty-six male Wistar rats were allocated into control, arsenic (15 mg/kg; orally), arsenic (15 mg/kg) plus melatonin (10, 20, and 30 mg/kg; intraperitoneally), and melatonin alone (30 mg/kg) groups for 28 days. After the treatment period, the serum sample was separated to measure liver enzymes (AST and ALT). The liver tissue was removed and then histological alterations, oxidative stress markers, antioxidant capacity, the levels of Nrf2 and HO-1, apoptosis (Bcl-2, survivin, Mcl1, Bax, and caspase-3), and autophagy (Sirt1, Beclin-1, and LC3 II/I ratio) proteins, as well as the expression level of miR-34a, were evaluated on this tissue. Arsenic exposure resulted in the enhancement of serum AST, ALT, and substantial histological damage in the liver. Increased levels of malondialdehyde, a lipid peroxidation marker, and decreased levels of physiological antioxidants including glutathione, superoxide dismutase, and catalase were indicators of arsenic-induced oxidative damage. The levels of Nrf2, HO-1, and antiapoptotic proteins diminished, while proapoptotic and autophagy proteins were elevated in the arsenic group concomitant with a low level of hepatic miR-34a. The co-treatment of melatonin and arsenic reversed the changes caused by arsenic. These findings showed that melatonin reduced the hepatic damage induced by arsenic due to its antioxidant and antiapoptotic properties as well as its regulatory effect on the miR-34a/Sirt1/autophagy pathway.

The Protective Effects of Trametes Versicolor on Arsenic-Induced Male Reproductive Toxicity through Regulation of Oxidative Stress: A Biochemical and Histopathological Survey

Exposure to arsenic is linked to a wide range of diseases, in particular male reproductive toxicities. Trametes versicolor is a traditional medicinal fungus with a remarkable potential for antioxidant activity. The purpose of this study was to examine the ameliorating effects of water and methanol extracts of T. versicolor on arsenic-induced male reproductive toxicities via the abrogation of oxidative stress. The mice were divided as follows: control: normal saline, As: arsenic (15 mg/kg), WE: water extract (400 mg/kg), ME: methanol extract (400 mg/kg), As + WE: arsenic (15 mg/kg) + water extract (100, 200, 400 mg/kg), As + ME: arsenic (15 mg/kg) + methanol extract (100, 200, 400 mg/kg), and positive control: arsenic (15 mg/kg) + vitamin C (500 mg/kg). Animals were treated via the intraperitoneal route. About 24 hr later, the mice were euthanized, and oxidative stress parameters (reactive oxygen species [ROS], lipid peroxidation, glutathione concentration, protein carbonylation, glutathione peroxidase, and superoxide dismutase activity), histopathological changes and sperm parameters (count, motility, and morphology) were examined in the testicular tissue. Arsenic caused significant pathological changes in the testicular tissue and sperm morphology and significantly reduced sperm count and motility. Moreover, arsenic mediated oxidative stress via significant increases in ROS generation, lipid peroxidation, and protein carbonyl content, as well as significant depletion in glutathione concentration and superoxide dismutase and glutathione peroxidase activities. Although, coadministration of water and methanol extracts of T. versicolor at 200 and 400 mg/kg counteracted arsenic-induced oxidative and histopathological damages and improved sperm parameters. Our study indicated that T. versicolor ameliorated arsenic-induced testis toxicity and sperm dysfunction via attenuation of oxidative damage.

Determinants of oxidative stress among indigenous populations in Northern Laos: Trace element exposures and dietary patterns

ObjectivesTo investigate determinants of oxidative stress in an indigenous population, we examined associations of trace element exposures and dietary patterns with three oxidative stress-related biomarkers among indigenous populations in Northern Laos. MethodsThis cross-sectional study included 341 adults from three villages with different levels of modernization. We used three oxidative stress-related biomarkers: urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) and 8-isoprostane concentrations, which were measured using liquid chromatography-tandem mass spectrometry, and blood telomere lengths, which were measured using a quantitative polymerase chain reaction method. We used multilevel analysis to examine associations of urinary arsenic, cadmium, and selenium concentrations, their interaction terms, and wild-plant-food scores (principal component scores calculated from food consumption frequencies) with oxidative stress-related biomarkers. ResultsUrinary arsenic and cadmium concentrations were positively associated with urinary 8-isoprostane concentrations. Urinary selenium concentrations were positively associated with urinary 8-OHdG concentrations. Interaction terms ([arsenic or cadmium] × selenium) showed negative associations with urinary 8-OHdG and 8-isoprostane concentrations, respectively. Urinary cadmium concentrations were negatively associated with telomere lengths. Wild-plant-food scores did not exhibit associations with oxidative stress-related biomarkers. ConclusionOur findings imply that exposure to arsenic and cadmium is associated with greater oxidative lipid damage, whereas selenium may attenuate arsenic-induced oxidative DNA damage and cadmium-induced oxidative lipid damage. Cadmium exposure may accelerate telomere attrition. Trace element exposure may be a determinant of oxidative stress among indigenous populations in Northern Laos.

Effects of exogenous salicylic acid on alleviation of arsenic-induced oxidative damages in rice

Salicylic acid (SA) is a phenolic phytohormone that plays a vital role in plant development and mediates plant responses to plenty of adversity including arsenic (As) stress. The effects of exogenous addition of SA on As tolerance and As accumulation were assessed in two cultivars of rice (Oryza sativa L.) Nipponbare and Zhongzao 39, hydroponically grown with Kimura B nutrient solution under arsenite [As (III)] and dimethylarsonic acid (DMA) exposure. In the second ex-periment, the influence of soaking seed with SA on As uptake and As damages was investigated in rice (cv. Nipponbare) exposed to As (III) and DMA. The results showed that exogenous addition of SA significantly decreased the concentrations of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in both As (III)- and DMA-stressed rice, indicating that SA alleviates As-induced oxidative damages in rice. SA increased the activity of antioxidant enzymes and, moreover, increased the relative amount of glutathione (GSH) and ascorbate (ASA) by accelerating the GSH-ASA circle system. Exogenous addition of SA significantly decreased the As concentration in both roots and shoots of rice under As(III) stress by influencing the expression of genes encoding As transporters, viz. OsLsi1, OsLsi2. The addition of SA significantly decreased the As content in shoots under DMA stress, which may be related to the expression of OsPTR7 involved in shoot xylem unloading. This finding may foster a novel perspective for reducing As accumulation in rice grains.

Betaine protects against sodium arsenite-induced diabetes and hepatotoxicity in mice.

Epidemiological evidence has associated chronic exposure to inorganic arsenic with an increased occurrence of glucose intolerance and diabetes mellitus. Furthermore, inorganic arsenic induces oxidative stress in organs such as the liver. Betaine, as a methyl donor, plays a pivotal role in homocysteine metabolism. Betaine has antioxidant and anti-inflammatory properties. Therefore, the aim of this study was to evaluate the effects of betaine against sodium arsenite-induced diabetes and hepatotoxicity in mice. Forty-eight male mice were divided into 6 groups of 8. Group 1, received distilled water every day for 4weeks by gavage. Group 2 received 500mg/kg betaine every day for 4weeks by gavage. Group 3 was given 10mg/kg NaAsO2 every day for 4weeks by gavage. Groups 4, 5, and 6 were co-treated with 125, 250, and 500mg/kg betaine half an hour before NaAsO2 (10mg/kg), respectively, daily for up to 4weeks by gavage. After 28days of the study, the mice were fasted overnight and on day 29, fasting blood glucose was measured and glucose tolerance test was performed. On day 30, the mice were anesthetized and a blood sample was taken from the heart. Serum factors (alanine aminotransferase, aspartate transaminase, and alkaline phosphatase activities), oxidative stress factors (malondialdehyde and glutathione levels, and the activity of superoxide dismutase, glutathione peroxidase, and catalase enzymes) and hepatic inflammatory factors (nitric oxide and tumor necrosis factor α) were measured. Histopathological studies were also performed on the liver and pancreas. In this study, it was shown that arsenic causes glucose intolerance, and oxidative/inflammatory hepatic damage. Co-administration of betaine prevents hepatotoxicity and glucose intolerance induced by arsenic in mice. Co-treatment of betaine with arsenic improved glucose intolerance and protected the liver against arsenic induced-oxidative damage and inflammation. Betaine at the dose of 500mg/kg showed better results than the other doses. Accordingly, betaine can be suggested as a therapeutic agent against diabetogenic and hepatotoxic effects of arsenic.

SAM and folic acid prevent arsenic-induced oxidative and nitrative DNA damage in human lymphoblast cells by modulating expression of inflammatory and DNA repair genes

Growing evidence suggests that arsenic exposure increases the risk of developing a variety of inflammation-associated chronic diseases and cancers. Our previous study revealed that increased transcript levels of inflammatory genes (i.e. COX2, EGR1, and SOCS3) coupled with hypomethylation of the promoter regions of these genes was associated with increased DNA damage in arsenic-exposed newborns through their early childhood. This study further investigated the ability of the methyl group donors, S-adenosyl methionine (SAM) and folic acid, to prevent promoter hypomethylation that results in decreased mRNA expression of inflammatory genes (COX2, EGR1, and SOCS3), and a reduction in arsenic-induced oxidative and nitrative DNA damage in human lymphoblast cells. Pretreatment with SAM (100 nM, 2 days) increased promoter methylation, reduced the mRNA levels of these inflammatory genes, and decreased both 8-hydroxydeoxyguanosine (8-OHdG) and 8-nitroguanine levels by 50% (p < 0.01) in arsenic-treated cells. In addition, pretreatment with folic acid (10 μM, 7 days), a micronutrient, led to a significant increase in promoter methylation associated with the reduction in mRNA levels of these inflammatory genes and decreased levels of 8-OHdG and 8-nitroguanine by 80% and 90% (p < 0.01), respectively, compared with arsenic treatment alone. Moreover, pretreatments with these methyl group donors increased mRNA expression of an antioxidant defense regulator (Nrf2) and DNA repair genes (hOGG1, XRCC1, and PARP1). This study shows for the first time that SAM or folic acid supplementation can prevent arsenic-induced oxidative and nitrative DNA damage. This suggests the potential use of SAM or folic acid for prevention of arsenic toxicity in human populations.

Aquoeus Extracts of Daucus Carota (Linn) Protected the Postnatal Developing Cerebellum of Wistar Rats Against Arsenic-Induced Oxidative Stress.

The neuroprotective effects of the aqueous extract of Daucus carota (Dc) tuber against arsenic-induced oxidative damage on the developing cerebellum of Wistar rats were studied. Twenty-five pregnant rats (110-200g) were divided into five groups (n=5) - control received distilled water; Arsenic (As); Dc (200mg/kg); Dc (200mg/kg) +As; Vitamin C (Vc) (100mg/kg) +As. The pregnant rats in all the groups were treated orally from the first day of pregnancy to postnatal day 21. The Dc extract and Vc were administered one hour before the administration of As. Body weight of the pups on days 1, 7, 14, 21 and 28 were recorded, while neurobehavioural (forelimb grip strength and negative geotaxis) tests were done on day 21 pups. The rats were sacrificed and cerebellar tissues were collected for oxidative stress, histological (H and E), and immunohistochemical studies.Decreased forelimb grip strength, increased lipid peroxidation and decreased glutathione, glutathione peroxidase, catalase and superoxide dismutase was observed in the As group compared with the control and other treated groups. Histologically, the cerebellar cortex of the As pups showed persistent external granular layer (EGL) on postnatal day 21, reduced thickness of the molecular layer (ML) on postnatal day 28, pyknotic and depleted Purkinje cells compared with the control and other treated rats. Immunohistochemical evaluations of the cerebellar cortex showed astroliosis in the As-treated group on day 21 pups compared with the control and other treated groups. Aqueous extracts of Daucus carota and Vitamin C reversed the toxicity caused by arsenic. From the results of the study, arsenic-induced oxidative stress with morphological alterations in the perinatal developing rat cerebellum. Extracts of Daucus carota exhibited antioxidant activity as such may be a potential neuroprotective agent.

Aquoeus Extracts of Daucus Carota (Linn) Protected the Postnatal Developing Cerebellum of Wistar Rats Against Arsenic-Induced Oxidative Stress

The neuroprotective effects of the aqueous extract of Daucus carota (Dc) tuber against arsenic-induced oxidative damage on the developing cerebellum of Wistar rats were studied. Twenty-five pregnant rats (110-200g) were divided into five groups (n=5) – control received distilled water; Arsenic (As); Dc (200mg/kg); Dc (200mg/kg) +As; Vitamin C (Vc) (100mg/kg) +As. The pregnant rats in all the groups were treated orally from the first day of pregnancy to postnatal day 21. The Dc extract and Vc were administered one hour before the administration of As. Body weight of the pups on days 1, 7, 14, 21 and 28 were recorded, while neurobehavioural (forelimb grip strength and negative geotaxis) tests were done on day 21 pups. The rats were sacrificed and cerebellar tissues were collected for oxidative stress, histological (H and E), and immunohistochemical studies. Decreased forelimb grip strength, increased lipid peroxidation and decreased glutathione, glutathione peroxidase, catalase and superoxide dismutase was observed in the As group compared with the control and other treated groups. Histologically, the cerebellar cortex of the As pups showed persistent external granular layer (EGL) on postnatal day 21, reduced thickness of the molecular layer (ML) on postnatal day 28, pyknotic and depleted Purkinje cells compared with the control and other treated rats. Immunohistochemical evaluations of the cerebellar cortex showed astroliosis in the As-treated group on day 21 pups compared with the control and other treated groups. Aqueous extracts of Daucus carota and Vitamin C reversed the toxicity caused by arsenic. From the results of the study, arsenic-induced oxidative stress with morphological alterations in the perinatal developing rat cerebellum. Extracts of Daucus carota exhibited antioxidant activity as such may be a potential neuroprotective agent.

Resveratrol protects against inorganic arsenic-induced oxidative damage and cytoarchitectural alterations in female mouse hippocampus

Prolonged inorganic arsenic (iAs) exposure is widely associated with brain damage particularly in the hippocampus via oxidative and apoptotic pathways. Resveratrol (RES) has gained considerable attention because of its benefits to human health. However, its neuroprotective potential against iAs-induced toxicity in CA1 region of hippocampus remains unexplored.Therefore, we investigated the neuroprotective efficacy of RES against arsenic trioxide (As2O3)-induced adverse effects on neuronal morphology, apoptotic markers and oxidative stress parameters in mouse CA1 region (hippocampus).Adult female Swiss albino mice of reproductive maturity were orally exposed to either As2O3 (2 and 4 mg/kg bw) alone or in combination with RES (40 mg/kg bw) for a period of 45 days. After animal sacrifice on day 46, the perfusion fixed brain samples were used for the observation of neuronal morphology and studying the morphometric features. While the freshly dissected hippocampi were processed for biochemical estimation of oxidative stress markers and western blotting of apoptosis-associated proteins.Chronic iAs exposure led to significant decrease in Stratum Pyramidale layer thickness along with reduction in cell density and area of Pyramidal neurons in contrast to the controls. Biochemical analysis showed reduced hippocampal GSH content but no change in total nitrite (NO) levels following iAs exposure. Western blotting showed apparent changes in the expression levels of Bax and Bcl-2 proteins following iAs exposure, however the change was statistically insignificant. Contrastingly, iAs +RES co-treatment exhibited substantial reversal in morphological and biochemical observations.Together, these findings provide preliminary evidence of neuroprotective role of RES on structural and biochemical alterations pertaining to mouse hippocampus following chronic iAs exposure.

The Use of Topical Estrogen Supplementation to Combat Arsenic‐Induced Redox Stress in Ovariectomized Murine Models

Background Chronic environmental arsenic exposure is a health hazard that can lead to cancer, cardiovascular disease and diabetes. Arsenic exposure stimulates the production of reactive oxygen species (ROS) increasing systemic oxidative stress. Estrogen contains antioxidant properties that assist in the upregulation of various antioxidant enzymes, including glutathione. The present study investigated the effects of estrogen as a therapeutic agent to combat oxidative stress caused by arsenic exposure in ovariectomized murine models. Methods Ovariectomized mice (10 weeks old) (n=24) were randomized to four treatment groups (CON= control, CE=control + estrogen, ARS=arsenic, AE= arsenic + estrogen). Arsenic treated groups (ARS & AE) were exposed to controlled arsenic levels (10µM) via drinking water for 3-wks. Following the arsenic exposure, estrogen supplementation (0.75 µg/g of 0.06% Estradiol Benzoate) was applied topically to respective treatment groups (CE & AE). One week following the first estrogen dosage (3 total doses), mice were euthanized, and skeletal muscle and cardiac tissue were collected. Oxidized and reduced glutathione and catalase (CAT) activity were measured in skeletal muscle. Superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were measured in cardiac tissue. A One-way ANOVA was performed with post-hoc comparisons using Least Significant Differences (LSD) for pairwise comparisons. Results For reduced glutathione levels, CE was significantly higher than CON (p = 0.004) and ARS (p = 0.001). CE was also significantly higher than all other groups for GSSG (p < 0.05) for all pairwise comparisons). Catalase activity was significantly higher in AE compared to CON (p = 0.032), and CE (p = 0.001), and ARS (p = 0.009). Superoxide dismutase (SOD) and thioredoxin reductase (TrxR) levels did not differ across treatment groups. Conclusion Estrogen treatment increased total glutathione content in the CE group indicated by the high levels of GSH as well as GSSG. Estrogen treatment combined with the arsenic exposure also increased catalase activity levels pointing to higher levels of hydrogen peroxide in the AE group, although this was not measured directly. Taken together, these data suggest estrogen may increase antioxidant capacity by elevating total glutathione and catalase content within skeletal muscle. These data indicate that estrogen alone and estrogen in the presence arsenic may play a role in modulating antioxidant activity in arsenic-induced oxidative damage and may be a potential treatment option for individuals suffering from environmental arsenic exposure.

Arsenic exposure through drinking Water and oxidative stress Status: A cross-sectional study in the Ayeyarwady region, Myanmar

Arsenic is a well-known toxic heavy metal that is naturally dispersed in groundwater. Whereas arsenic is widely accepted to be involved in oxidative stress damage, little is known about arsenic-induced oxidative damage in relationship to contaminated drinking water as a source. The aim of this study was to determine the association between arsenic exposure through drinking water and oxidative stress status by measuring levels of urinary 8-hydroxydeoxyguanosine (8−OHdG) as a biomarker of oxidative stress damage in a Myanmar population. A questionnaire-based survey and drinking water and urine sampling (n = 198) were performed to assess the association between arsenic exposure and urinary 8−OHdG concentration in the Ayeyarwady Region, Myanmar. Urinary arsenic concentrations were significantly correlated with drinking water arsenic concentrations (Spearman’s rho = 0.32, p < 0.001). Multivariate linear regression analysis showed that higher urinary arsenic concentrations were significantly associated with higher 8−OHdG concentrations (coefficient = 0.09, 95% confidence interval, 0.03 – 0.15; p = 0.002). The present study identified that exposure to arsenic through drinking water could induce an increase in the urinary 8−OHdG concentration, reflecting increased oxidative DNA damage. These findings provide evidence that may explain the role of arsenic-induced oxidative stress in the pathophysiology of arsenic-induced diseases including cancers.

Sulforaphane Potentially Ameliorates Arsenic Induced Hepatotoxicity in Albino Wistar Rats: Implication of PI3K/Akt/Nrf2 Signaling Pathway.

Agriculture and industrial expansion in recent years have resulted in the undue discharge of arsenic into the environment, building arsenic toxicity a major worldwide anxiety. Oxidative stress is considered as the most conspicuous effect of arsenic toxicity. The current study was designed to evaluate the protective ability of sulforaphane (SFN) against arsenic (As) induced hepatotoxicity by activation of PI3K induced Akt and Nrf2 mediated signaling pathway. For this purpose, male Wistar rats were randomly distributed into 6 groups of 8 rats each: control, Arsenic (5mg/kg BW), SFN plus Arsenic (20, 40, 80 mg/kg BW; 5mg/kg BW) and Vit. C plus Arsenic (100mg/kg BW; 5mg/kg BW). In this study, we have used spectrophotometry for enzymatic antioxidant assays, western blotting and PCR for protein and gene expression. Microtome for histological study. The arsenic-induced oxidative damage was confirmed by a significant (p<0.05) increase in the levels of ALAD, As concentration and depletion in the antioxidant content. Furthermore, Arsenics treatments significantly (p<0.05) increased the pro-apoptotic marker (Bax) and DNA damage, with decreased Nrf2 protein responsible for liver protection. However, pretreatment with SFN significantly (p<0.05) decreased the levels of ALAD, Arsenic concentration, and brought antioxidant enzymes into normal levels. This was accomplished by inhibition of apoptotic markers via activation of PI3K, Akt and Nrf2 mediated signaling pathway as evident from western blotting and PCR techniques. Moreover, SFN pretreatment shield the liver histoarchitecture observed in Arsenic treated groups suggesting prevention of liver toxicity via PI3K/Akt mediated Nrf2 signaling pathways and could possibly provide a protection against Arsenic induced hepatic burden.

Proanthocyanidins Antagonize Arsenic-Induced Oxidative Damage and Promote Arsenic Methylation through Activation of the Nrf2 Signaling Pathway.

Purpose To investigate the effects of grape seed proanthocyanidin extract (GSPE) on oxidative damage and arsenic (As) methylation and to clarify the role of Nrf2 in the process. Methods L-02 cells were treated with arsenic (25 μM) and GSPE (10, 25, and 50 mg/L) for 24 h. Cell viability was analyzed by MTT assay. Cell apoptosis and ROS fluorescence were detected by flow cytometry. Oxidative stress marker levels were measured using commercial kits. mRNA and protein expression were detected by qRT-PCR and western blotting. The cellular concentrations of methylation products were measured by HPLC-HGAFS. Arsenic methylation ability of cells was determined. Results Cell survival rate was significantly lower in the As group than in the control group (P < 0.05), while cell apoptosis increased and the number of apoptotic cells decreased gradually after GSPE intervention. Superoxide dismutase, glutathione, and sulfhydryl levels in the intervention group were significantly higher (P < 0.05), while MDA and ROS levels were significantly lower (P < 0.05) than those in the As group. The mRNA and protein expression of Nrf2, HO-1, NQO1, and glutathione-S-transferase increased in the As + GSPE group compared with that in the As group (P < 0.05). GSPE significantly increased methylated As level, primary methylation index, secondary methylation index, average growth rate of methylation, and average methylation speed compared with the GSPE untreated group (P < 0.05). After Nrf2 inhibition, the effect of GSPE decreased significantly. Conclusion GSPE activates the Nrf2 signaling pathway to antagonize As-induced oxidative damage and to promote As methylation metabolism. Therefore, GSPE may be a potential agent for relieving As-induced hepatotoxicity.

Transcriptional regulation of Fe[sbnd]S biogenesis genes: A possible shield against arsenate toxicity activated by Yap1

In the eukaryotic model yeast Saccharomyces cerevisiae, arsenic (As) detoxification is regulated by two transcriptional factors, Yap8 and Yap1. Yap8 specifically controls As extrusion from the cell, whether Yap1 avoids arsenic-induced oxidative damages. Accordingly, cells lacking both Yap1 and Yap8 are more sensitive to arsenate than cells lacking each regulator individually. Strikingly enough, the same sensitivity pattern was observed under anoxia, suggesting that Yap1 role in As detoxification might not be restricted to the regulation of the oxidative stress response. This finding prompted us to study the transcriptomic profile of wild-type and yap1 mutant cells exposed to arsenate. Interestingly, we found that, under such conditions, several genes involved in the biogenesis of FeS proteins were upregulated in a Yap1-dependent way. In line with this observation, arsenate treatment decreases the activity of the mitochondrial aconitase, Aco1, an FeS cluster-containing enzyme, this effect being even more pronounced in the yap1 mutant. Reinforcing the relevance of FeS cluster biogenesis in arsenate detoxification, the overexpression of several ISC and CIA machinery genes alleviates the deleterious effect of arsenate caused by the absence of Yap1 and Yap8.Altogether our data suggest that the upregulation of FeS biogenesis genes regulated by Yap1 might work as a cellular shield against arsenate toxicity.

Acetyl-l-carnitine attenuates arsenic-induced liver injury by abrogation of mitochondrial dysfunction, inflammation, and apoptosis in rats

Industrial and agricultural developments in recent years have resulted in the excessive discharge of arsenic into the environment, making arsenic toxicity a major worldwide concern. Oxidative stress is considered the primary mechanism for arsenic toxicity. The main objective of this study was to evaluate acetyl-l-carnitine’s (ALC) protective ability against the arsenic-induced hepatotoxicity. For this purpose, male Wistar rats were distributed randomly into 5 groups of 8 rats each: control, arsenic (5 mg/kg) and arsenic plus ALC (5 mg/kg; 100, 200, 300 mg/kg). The animals were gavaged for 21 consecutive days. Liver tissue samples were extracted 24 h after the last treatment and were later analyzed for biochemical and histological alterations. The arsenic-induced oxidative damage was confirmed by elevation of malondialdehyde (MDA), a lipid peroxidation byproduct, as well as depletion in physiological antioxidant content such as superoxide dismutase (SOD) and catalase (CAT). Furthermore, alterations in mitochondrial functions including a significant decrease of mitochondrial outer membrane potential and reactive oxygen species (ROS) generation increase, mitochondrial swelling, release of cytochrome c and consequent activation of caspase-3 and caspase-9 and initiation of apoptosis, was observed following arsenic administration. Moreover, the inflammation was confirmed by the overexpression of inflammatory mediators such as NF-ĸB and IL-1 and IL-6. The present study demonstrated that ALC ameliorates arsenic-induced oxidative damage, mitochondrial dysfunction, apoptosis, inflammation and histological damage. ALC’s protective features against arsenic hepatotoxicity may be due to this agent’s antioxidant and anti-inflammatory properties as well as its stabilizing effects on mitochondrial function.

Methyl jasmonate alleviates arsenic-induced oxidative damage and modulates the ascorbate–glutathione cycle in oilseed rape roots

Methyl jasmonate (MJ) is an important plant growth regulator, involves in various physiological processes of plants. In the present study, role of MJ in tolerance to oilseed rape (Brassica napus L.) roots under arsenic (As) stress was investigated. The treatments were comprised of three MJ doses (0, 0.1, and 1 µM) and two levels of As (0 and 200 µM). Arsenic stress resulted in oxidative damage as evidenced by decreased root growth and enhanced reactive oxygen species and lipid peroxidation. However, plants treated with MJ decreased the H2O2 and O2 ·− contents in roots and have higher antioxidant activities. Importantly, results showed that MJ enhanced the redox states of AsA and GSH, and the related enzymes involved in the AsA–GSH cycle. Moreover, MJ also induced the secondary metabolites related enzymes (PAL and PPO) activities, under As stress. PAL and PPO expression was further increased by MJ application in the roots of B. napus under As stress. MJ also reduced the total As content compared with As alone treated plants. These findings suggest the role of MJ in mitigation of the As-induced oxidative damage by regulating AsA and GSH redox states and by reducing As uptake in both cultivars.

Effects of Nrf2 deficiency on arsenic metabolism in mice

Inorganic arsenic (iAs) is a known toxicant and carcinogen. Worldwide arsenic exposure has become a threat to human health. The severity of arsenic toxicity is strongly correlated with the speed of arsenic metabolism (methylation) and clearance. Furthermore, oxidative stress is recognized as a major mechanism for arsenic-induced toxicity. Nuclear factor-E2-related factor 2 (Nrf2), a key regulator in cellular adaptive antioxidant response, is clearly involved in alleviation of arsenic-induced oxidative damage. Multiple studies demonstrate that Nrf2 deficiency mice are more vulnerable to arsenic-induced intoxication. However, what effect Nrf2 deficiency might have on arsenic metabolism in mice is still unknown. In the present study, we measured the key enzymes involved in arsenic metabolism in Nrf2-WT and Nrf2-KO mice. Our results showed that basal transcript levels of glutathione S-transferase omega 2 (Gsto2) were significantly higher and GST mu 1 (Gstm1) lower in Nrf2-KO mice compared to Nrf2-WT control. Arsenic speciation and methylation rate in liver and urine was then studied in mice treated with 5mg/kg sodium arsenite for 12h. Although there were some alterations in arsenic metabolism enzymes between Nrf2-WT and Nrf2-KO mice, the Nrf2 deficiency had no significant effect on arsenic methylation. These results suggest that the Nrf2-KO mice are more sensitive to arsenic than Nrf2-WT mainly because of differences in adaptive antioxidant detoxification capacity rather than arsenic methylation capacity.

Studies on curative efficacy of monoterpene eugenol on anti- leukemic drug arsenic trioxide induced cardiotoxicity

BackgroundArsenic trioxide (As2O3) is emerging as a frontline agent for the treatment of acute promyelocytic leukemia (APL) but the therapeutic application is limited by its toxicity. QT prolongation, torsades de pointes and sudden cardiac death have been implicated in the As2O3 therapy. So eugenol is a monoterpene compound is well known for its antioxidant properties and protective effect on the cardiovascular system. ObjectiveIn this study, the cardioprotective effect of eugenol on cardiac electrical conductivity, tissue electrolytes, myocardial markers, antioxidant system, lipid peroxidation and nitric oxide production was investigated in male Wistar rats treated with arsenic trioxide. ResultsThe Inductively coupled plasma emission spectroscopic (ICP-OES) analysis pointed out the accumulation of arsenic in heart tissue. The rats administered with arsenic trioxide (4mg/kg body wt) exhibited myocardial damage that was manifested by the elevation of cardiac markers (LDH, CK-MB) enzymes and deterioration in the antioxidant enzymes (GSH, GST, GPx). Combination treatment with eugenol (5mg/kg of body wt) upholds the tissue antioxidant level, Na+/K+ − ATPase and Ca2+− ATPase activity and brings the cytosolic Ca2+, K+ and Na + levels near to normal value. Conjoined therapy with eugenol ameliorated the membrane peroxidation, restored the normal heart rate and rectified the prolongation of QT interval in the electrocardiogram. Histological examination of cardiac segments also supported the beneficial role of eugenol against arsenic-induced oxidative damages. ConclusionOur in vivo experimental findings suggest that monoterpenoid eugenol could be a potent and novel cytoprotective agent of clinical application against As2O3 induced cardiotoxicity.

Pomegranate protects against arsenic-induced p53-dependent ROS-mediated inflammation and apoptosis in liver cells

Molecular mechanisms involved in arsenic-induced toxicity are complex and elusive. Liver is one of the most favored organs for arsenic toxicity as methylation of arsenic occurs mostly in the liver. In this study, we have selected a range of environmentally relevant doses of arsenic to examine the basis of arsenic toxicity and the role of pomegranate fruit extract (PFE) in combating it. Male Swiss albino mice exposed to different doses of arsenic presented marked hepatic injury as evident from histological and electron microscopic studies. Increased activities of enzymes alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase corroborated extensive liver damage. It was further noted that arsenic exposure initiated reactive oxygen species (ROS)-dependent apoptosis in the hepatocytes involving loss of mitochondrial membrane potential. Arsenic significantly increased nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear factor-κB (NF-κB), coupled with increase in phosphorylated Iκ-B, possibly as adaptive cellular survival strategies. Arsenic-induced oxidative DNA damage to liver cells culminated in p53 activation and increased expression of p53 targets like miR-34a and Bax. Pomegranate polyphenols are known to possess remarkable antioxidant properties and are capable of protecting normal cells from various stimuli-induced oxidative stress and toxicities. We explored the protective role of PFE in ameliorating arsenic-induced hepatic damage. PFE was shown to reduce ROS generation in hepatocytes, thereby reducing arsenic-induced Nrf2 activation. PFE also inhibited arsenic-induced NF-κB-inflammatory pathway. Data revealed that PFE reversed arsenic-induced hepatotoxicity and apoptosis by modulating the ROS/Nrf2/p53–miR-34a axis. For the first time, we have mapped the possible signaling pathways associated with arsenic-induced hepatotoxicity and its rescue by pomegranate polyphenols.

Promising anti-oxidative therapeutic potentials of edible freshwater snail Bellamya bengalensis extract against arsenic-induced rat hepatic tissue and DNA damage

Epidemiological data suggest that arsenic ultimately results in cancer in different parts of the body. Several synthetic therapeutic agents manifest inadequate potency with severe side effects against arsenic toxicity. The flesh of B. bengalensis , has long been used as an ethno-medicine in case of arthritis, blood-impurities, impaired immune system, conjunctivitis and liver anomalies. This potent organism might be a natural choice against arsenic and several other toxicities. Our earlier studies on arsenic-exposed human can correlate carcinogenesis with DNA-damage. In an attempt to investigate the possible protective and therapeutic effect against arsenic induced hepatotoxicity, the extract of B. bengalensis was tested in arsenic intoxicated rat model. The time- and dose-dependent effect of arsenic toxicity was also tested in B. bengalensis . Sodium-meta-arsenite NaAsO 2 (0.6 ppm/100g bw/day for 28 days, as earlier reported) was treated alone or in combination with the B. bengalensis water extract (BBE, 100 mg/100g bw) to rat and compared with vehicle treated control. In a separate experiment, the B. bengalensis was exposed to high concentration of NaAsO 2 contaminated water (5 to 20 ppm for 1 to 9 days) in laboratory condition and their DNA quality was evaluated in relation to its possible oxidative threat. Any concentration of arsenic was incapable to initiate a significant DNA damage in B. bengalensis . Lipid peroxidation was increased in arsenic exposed B. bengalensis after longer duration of its exposure. Increase in reduced antioxidant like non-protein-soluble thiol (NPSH) is concordant with the decrease in lipid peroxidation and DNA stability in this organism. In rat experiment, the BBE supplementation strongly prevented arsenic-induced oxidative, necrotic and apoptotic damages to liver tissue/DNA by strengthening antioxidant systems, which has been shown in hepatic DNA-fragmentation, comet-assay, histo-architecture (hematoxylin/eosin), alkaline-phosphatase, serum-glutamate-pyruvate-transaminase and lactate-dehydrogenase (tissue-degeneration-marker) results. Only arsenic exposure decreased hepatic superoxide-dismutase (SOD) in-vivo and in-vitro (H 2 O 2 /arsenite redox-stress to dialyzed and concentrated, 6-8kd cutoff-Millipore liver cytosolic SOD), catalase, xanthine-oxidase, lactoperoxidase activities and the level of NPSH with a concomitant increase in malondialdehyde resulting in mutagenic DNA-breakage and apoptotic liver damage which has been decisively restrained in B. bengalensis extract. The present investigation offers strong evidence on the hepato-protective and medicinal efficiencies of BBE against oxidative stress induced by arsenic.