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Eriocheir sinensis, a key species in China's freshwater aquaculture, is threatened by various diseases, which were verified to be closely associated with oxidative stress. This study aimed to investigate the response of E. sinensis to hydrogen peroxide (H2O2)-induced oxidative stress to understand the biological processes behind these diseases. Crabs were exposed to different concentrations of H2O2 and their antioxidant enzyme activities and gene expressions for defense and immunity were measured. Results showed that activities of antioxidant enzymes—specificallysuperoxide dismutase (SOD), catalase (CAT), total antioxidant capacity(T-AOC), glutathione (GSH), and glutathione peroxidase (GSH-Px)—varied with exposure concentration and duration, initially increasing then decreasing. Notably, SOD, GSH-Px, and T-AOC activities dropped below control levels at 96 h. Concurrently, oxidative damage markers, including malondialdehyde (MDA), H2O2, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels, increased with exposure duration. The mRNA expression of SOD, CAT, and GSH-Px also showed an initial increase followed by a decrease, peaking at 72 h. The upregulation of phenoloxidaseloxidase (proPO) and peroxinectin (PX) was also detected, but proPO was suppressed under high levels of H2O2. Heat shock protein 70 (HSP70) expression gradually increased with higher H2O2 concentrations, whereas induced nitrogen monoxide synthase (iNOS) was upregulated but decreased at 96 h. These findings emphasize H2O2's significant impact on the crab's oxidative and immune responses, highlighting the importance of understanding cellular stress responses for disease prevention and therapy development. [ABSTRACT FROM AUTHOR]

Background: The anti-infammatory and cardioprotective effect of acetylcholine (ACh) has been reported; nevertheless, whether and how ACh exhibits an antioxidant property against ischemia/reperfusion (I/R)-induced oxidative stress remains obscure. Methods: In the present study, H9c2 rat cardiomyocytes were exposed to hypoxia/reoxygenation (H/R) to mimic I/R injury. We estimated intracellular different sources of reactive oxygen species (ROS) by measuring mitochondrial ROS (mtROS), mitochondrial DNA (mtDNA) copy number, xanthine oxidase (XO) and NADPH oxidase (NOX) activity and expression of rac 1. Cell injury was determined by lactate dehydrogenase (LDH) release and cleaved caspase-3 expression. The siRNA transfection was performed to knockdown of M2 acetylcholine receptor (M2 AChR) expression. Results: 12-h hypoxia followed by 2-h reoxygenation resulted in an abrupt burst of ROS in H9c2 cells. Administration of ACh reduced the levels of ROS in a concentration-dependent manner. Compared to the H/R group, ACh decreased mtROS, recovered mtDNA copy number, diminished XO and NOX activity, rac 1 expression as well as cell injury. Co- treatment with atropine rather than hexamethonium abolished the antioxidant and cardioprotective effect of ACh. Moreover, knockdown of M2 AChR by siRNA showed the similar trends as atropine co-treatment group. Conclusions: ACh inhibits mitochondria-, XO- and NOX-derived ROS production thus protecting H9c2 cells against H/R-induced oxidative stress, and these benefcial effects are mainly mediated by M2 AChR. Our findings suggested that increasing ACh release could be a potential therapeutic strategy for treatment and prevention of I/R injury. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: The white shrimp Litopenaeus vannamei is one of the most important economic species in shrimp farming and are frequently exposed to multiple stressors (including temperature) in aquaculture. The differential expression of seven genes encoding antioxidant enzymes and stressor biomarkers was investigated by real-time quantitative PCR in haemocytes and hepatopancreas and gill extracted from L. vannamei following acute temperature stress. Temperature stress induced CAT, GST, ferritin and HSP60 gene expression in gills. Western blot results also revealed that HSP60 was induced in the same tissue after acute temperature stress. Heat stress resulted in an increase in most examined genes in haemocytes (excluding HSP60) suggesting haemocytes may be an early response tissue in acute temperature stress. GST was significantly up-regulated in haemocytes (to up to 16.4 fold at 22°C and 71.8 fold at 28°C, respectively) during exposure to heat stress. In addition, MnSOD was more strongly induced in haemocytes and hepatopancreas (to up to 273.8 fold and 115.8 fold, respectively) after exposure to 28°C from 15°C implying their important role in antioxidant protection in response to heat stress. The transcriptional responses of these genes to temperature stress will provide the basis for a multi-biomarker system that could be used for the biomonitoring of aquatic environments. [ABSTRACT FROM AUTHOR]

Abstract: The ROS production, the percentage of dead and damaged haemocytes, the DNA Olive Tail Moment (OTM) value and the gene expression of manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx) and thioredoxin (TRx), were studied in the Pacific white shrimp, Litopenaeus vannamei, when exposed to acute pH stress. The increased ROS production in haemocytes and the increased OTM value in both the haemocytes and the hepatopancreas cells suggest that oxidative damage occurred in shrimp exposed to pH 5.6 and pH 9.3, with apoptosis, mainly being associated with excess Ca2+influx and changes in cell viability. Acid and alkaline pH-induced DNA damage was time dependent in the haemocytes and the hepatopancreas cells. The concentration of intracellular free calcium [Ca2+] i after different pH treatments increased significantly over time, reaching its highest concentration after 12 h, but decreasing gradually to normal levels after 24 h. The [Ca2+] i content in shrimp cells when exposed to pH 9.3 and pH 5.6 for 12 h had increased by 58%–81%, compared with exposure to pH 7.4 (control). In addition, the gene expression of cMnSOD, CAT, GPx and TRx in the hepatopancreas of L. vannamei was induced by acid and alkaline pH stress, although there were differences in the expression response with respect to the duration of induction and the different pH treatments (acid or alkaline). Our results show that acidic or alkaline-induced oxidative stress may cause DNA damage, and cooperatively activate expression of CAT, GPx and TRx mRNA. Calcium ions appear to be important in mediating shrimp responses to pH stress. [Copyright &y& Elsevier]

Celastrus orbiculatus Thunb. is an ancient traditional Chinese herb with a long history of medicinal use. The ethyl acetate extract of Celastrus orbiculatus Thunb. (COE) has been shown to have anti-tumor effects in various preclinical studies. However, the anti-invasive and metastatic efficacy of COE in non-small cell lung cancer (NSCLC) and the mechanism by which COE regulates cellular oxidation levels are yet to be elucidated. To study the anti-dissemination effect of COE on NSCLC and to elucidate the molecular mechanism of COE in regulating cellular oxidation levels and its effect on lung cancer invasion and metastasis. CCK-8 assay was used to detect the toxic effects of COE on NSCLC. Transwell assay and high-content imaging was used to detect the Motility of NSCLC. Transmission electron microscopy and three-dimensional (3D) imaging of mitochondrial fluorescence were employed to detect the number and structure of mitochondria. JC-1 probe was used to detect the level of mitochondrial membrane potential. Firefly luciferase assay was used to detect the level of total intracellular ATP. MitoSox probe and DCFH-DA probe were applied to detect the level of reactive oxygen species (ROS) inside the mitochondria and the total intracellular ROS, respectively. Immunohistochemistry was used to detect protein expression in xenograft tumors. COE inhibited motility and induced DJ-1 downregulation in NSCLC at low toxic concentrations, and the antiseptic effect of COE was reduced significantly after the overexpression of DJ-1. COE induced structural disruption of mitochondria in NSCLC and accumulation of superoxide compounds, decreased the volume of membrane potential depolarization, and impaired energy production, ultimately leading to a large accumulation of ROS at the cellular level. The antioxidant acetylcysteine (NAC) significantly reversed the antiseptic capacity of COE. In a xenograft tumor model, protein expression of DJ-1, E-cadherin, N-cadherin, and MMP-2 in COE group was significantly changed compared to the model group. In the present study, COE inhibited NSCLC invasion and metastasis and was associated with the downregulation of DJ-1 and elevated ROS. COE-mediated downregulation of DJ-1 may be the primary cause of mitochondrial structural and functional dysfunction in NSCLC, eventually leading to ROS accumulation. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract: Methionine sulfoxide reductases A (MsrA) has been postulated to act as a catalytic antioxidant system involved in the protection of oxidative stress-induced cell injury. Recently, attention has turned to MsrA in coupling with the pathology of Parkinson''s disease, which is closely related to neurotoxins that cause dopaminergic neuron degeneration. Here, we firstly provided evidence that pretreatment with a natural polyphenol resveratrol (RSV) up-regulated the expression of MsrA in human neuroblastoma SH-SY5Y cells. It was also observed that the expression and nuclear translocation of forkhead box group O 3a (FOXO3a), a transcription factor that activates the human MsrA promoter, increased after RSV pretreatment. Nicotinamide , an inhibitor of silent information regulator 1 (SIRT1), prevented RSV-induced elevation of FOXO3a and MsrA expression, indicating that the effect of RSV was mediated by a SIRT1-dependent pathway. RSV preconditioning increased methionine sulfoxide(MetO)-reducing activity in SH-SY5Y cells and enhanced their resistance to neurotoxins, including chloramine-T and 1-methyl-4-phenyl-pyridinium. In addition, the enhancement of cell resistance to neurotoxins caused by RSV preconditioning can be largely prevented by MsrA inhibitor dimethyl sulfoxide. Our findings suggest that treatment with polyphenols such as RSV can be used as a potential regulatory strategy for MsrA expression and function. [Copyright &y& Elsevier]