Indicator Of Oxidative Damage Research Articles

Nox2 underpins microvascular inflammation and vascular contributions to cognitive decline

Chronic microvascular inflammation and oxidative stress are inter-related mechanisms underpinning white matter disease and vascular cognitive impairment (VCI). A proposed mediator is nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (Nox2), a major source of reactive oxygen species (ROS) in the brain. To assess the role of Nox2 in VCI, we studied a tractable model with white matter pathology and cognitive impairment induced by bilateral carotid artery stenosis (BCAS). Mice with genetic deletion of Nox2 (Nox2 KO) were compared to wild-type (WT) following BCAS. Sustained BCAS over 12 weeks in WT mice induced Nox2 expression, indices of microvascular inflammation and oxidative damage, along with white matter pathology culminating in a marked cognitive impairment, which were all protected by Nox2 genetic deletion. Neurovascular coupling was impaired in WT mice post-BCAS and restored in Nox2 KO mice. Increased vascular expression of chemoattractant mediators, cell-adhesion molecules and endothelial activation factors in WT mice post-BCAS were ameliorated by Nox2 deficiency. The clinical relevance was confirmed by increased vascular Nox2 and indices of microvascular inflammation in human post-mortem subjects with cerebral vascular disease. Our results support Nox2 activity as a critical determinant of VCI, whose targeting may be of therapeutic benefit in cerebral vascular disease.

An exopolysaccharide from Bacillus subtilis alleviates airway inflammatory responses via the NF-κB and STAT6 pathways in asthmatic mice.

Bacillus subtilis is an intestinal probiotic for immune homeostasis and its exopolysaccharide (EPS) is known to possess anti-inflammatory and antioxidant properties. The underlying mechanisms are not yet fully understood. In the present study, we investigated the effects of the EPS (50, 100, 200 mg/kg) on airway inflammation in asthmatic mice. Our results showed that EPS treatment of asthmatic mice significantly alleviated pathological damage in the lungs, remarkably decreased the counts of total inflammatory cells including lymphocytes, and eosinophils in the bronchoalveolar lavage fluid (BALF) and reduced indexes of oxidative damage. Moreover, the expression of type II T-helper cell (Th2) cytokines (interleukin- (IL)4 and -5) subsequent to EPS treatment was found to be dramatically down-regulated in a concentration-dependent manner. Additionally, the EPS treatment reduced JAK1, STAT6 and nuclear factor-κB (NF-κB) expression in the lungs of asthmatic mice. Taken together, these results suggest that the EPS from B. subtilis alleviates asthmatic airway inflammation, which involves the reduction in reactive oxygen species (ROS) and the down-regulation of the STAT6 and NF-κB inflammatory pathways, which can further reduce Th2 cytokine expression and eosinophilic inflammation. Thus, our findings provide a potential mechanism through which the EPS mitigates asthma, suggesting that the EPS could be a potential source of an anti-asthmatic drug.

Synthesis of diaryl urea derivatives and evaluation of their antiproliferative activities in colon adenocarcinoma

Colon cancer is one of the leading causes of cancer-related deaths today. Serious research for ideal chemotherapy continues today. In this context, newly synthesized molecules have an essential role in cancer treatment research. The effects of 5 diaryl urea derivatives synthesized within the scope of this study on the HT-29 colon cancer cell line were investigated for the first time in the literature by in-silico and in-vitro methods. Among the five compounds produced in the first stage of the study, DAU5 was found to have a cytotoxic effect on HT-29. However, it was determined that it did not show a serious cytotoxic effect on L929 (healthy fibroblast cell line) at the same doses. The efficacy of DAU5 was evaluated for expression of LC3B, an indicator of autophagy, and 8-OHdG, an indicator of oxidative stress-DNA damage. LC3B and 8-OHdG expression were lower in the DAU5 treatment group than in the control group. As a result, it was seen that DAU5, a diaryl urea derivative compound we synthesized in this study, has the potential to be a chemotherapeutic agent against colon cancer. However, our study needs to be supported by in vivo and clinical studies.

Determining the in vitro Anti-Aging Effect of the Characteristic Components from Eucommia ulmoides

To evaluate the potential anti-aging ability of Eucommia ulmoides, four characteristic components (chlorogenic acid, geniposidic acid, aucubin, quercetin) were selected to assess their effects on H2O2-induced oxidative damage model of human umbilical vein endothelial cell (HUVEC). Oxidative damage indexes, inflammatory factors, cell cycle, cell apoptosis, cell senescence, and their related proteins were analyzed by methyl thiazolyl tetrazolium (MTT) assay, enzyme-linked immunosorbent assay (ELISA), propidium iodide (PI) staining, annexin V-FITC/ PI double staining, SA β-galactosidase staining, and western blotting (WB). The results showed that H2O2- induced cell growth inhibition rate decreased as supplementation with characteristic components when compared to H2O2 group. Meanwhile, the contents of antioxidant indexes (reactive oxygen species, lactate dehydrogenase, molondialdehyde, superoxide dismutase, glutathione), inflammatory factors (nuclear factor kappa-B, intercellular cell adhesion molecule-1, vascular cell adhesion protein 1), and functional factors (NO, Endothelin-1) in characteristic components treated groups improved if comparison with H2O2 group, suggesting the characteristic components of E. ulmoides could alleviate H2O2-induced oxidative damage. Moreover, cell cycle, cell apoptosis, cell senescence, and their related proteins under characteristic components treatment exhibited a better effect than under H2O2 treatment, implying the characteristic components could participate in anti-aging via multiple pathways. These results manifested that the characteristic components of E. ulmoides posses the capacity of anti-aging, which provided a basis for investigating the anti-aging ability of E. ulmoides itself.

Sulforaphane-Mediated Nrf2 Activation Prevents Radiation-Induced Skin Injury through Inhibiting the Oxidative-Stress-Activated DNA Damage and NLRP3 Inflammasome.

This article mainly observed the protective effect of sulforaphane (SFN) on radiation-induced skin injury (RISI). In addition, we will discuss the mechanism of SFN’s protection on RISI. The RISI model was established by the irradiation of the left thigh under intravenous anesthesia. Thirty-two C57/BL6 mice were randomly divided into control group (CON), SFN group, irradiation (IR) group, and IR plus SFN (IR/SFN) group. At eight weeks after irradiation, the morphological changes of mouse skin tissues were detected by H&E staining. Then, the oxidative stress and inflammatory response indexes in mouse skin tissues, as well as the expression of Nrf2 and its downstream antioxidant genes, were evaluated by ELISA, real-time PCR, and Western blotting. The H&E staining showed the hyperplasia of fibrous tissue in the mouse dermis and hypodermis of the IR group. Western blotting and ELISA results showed that the inflammasome of NLRP3, caspase-1, and IL-1β, as well as oxidative stress damage indicators ROS, 4-HNE, and 3-NT, in the skin tissues of mice in the IR group were significantly higher than those in the control group (p < 0.05). However, the above pathological changes declined sharply after SFN treatment (p < 0.05). In addition, the expressions of Nrf2 and its regulated antioxidant enzymes, including CAT and HO-1, were higher in the skin tissues of SFN and IR/SFN groups, but lower in the control and IR groups (p < 0.05). SFN may be able to suppress the oxidative stress by upregulating the expression and function of Nrf2, and subsequently inhibiting the activation of NLRP3 inflammasome and DNA damage, so as to prevent and alleviate the RISI.

Altered Oxidative Status as a Cost of Reproduction in a Seabird with High Reproductive Costs.

AbstractLife history theory posits that reproduction is constrained by a cost of reproduction such that any increase in breeding effort should reduce subsequent survival. Oxidative stress refers to an imbalance between the prooxidant reactive oxygen species (ROS) and antioxidant defense. If not thwarted, ROS can cause damage to DNA, lipids, and proteins, potentially increasing the rate of senescence and decreasing cellular function. Reproduction is often associated with higher metabolic rates, which could increase production of ROS and lead to oxidative damage if the animal does not increase antioxidant protection. Thus, oxidative stress could be one mechanism creating a cost of reproduction. In this study we explored how reproduction may affect oxidative status differently between male and female thick-billed murres during early and late breeding seasons over three consecutive years. We manipulated breeding efforts by removing an egg from the nest of some individuals, which forced females to relay, and by handicapping other individuals by clipping wings. We measured total antioxidant capacity (TAC), uric acid (UA) concentration, and malondialdehyde (MDA; an index of lipid oxidative damage) concentration in blood plasma as well as activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in red blood cells. Oxidative status was highly variable across years, and year was consistently the most important factor determining oxidative status; inconsistent results in previous field studies may be because reproductive oxidative stress occurs only in some years. Females had lower SOD and GPx and higher MDA and TAC than males immediately after egg laying, suggesting that the cost of egg laying required investment in cheaper nonenzymatic antioxidant defenses that had lower capacity for defending against lipid peroxidation. Delayed birds had lower UA and lower SOD, GPx, and CAT activity compared with control birds. In conclusion, when reproductive costs increase via higher energy costs or longer breeding seasons, the oxidative status of both male and female murres deteriorated as a result of reduced antioxidant defenses.

Inhibitory effect of Idelalisib on selenite-induced cataract in Sprague Dawley rat pups

ABSTRACTPurposeThe aim of this study was to evaluate the therapeutic effect of Idelalisib, Apelisib and Copanlisib on 8-day-old cataract SD rat pups.Materials and methodsThe rat model induced by sodium selenate (Na2SeO3) was used in this study. Experimental animals were randomly divided into five groups with eight animals in each group. They were control group, Na2SeO3 group, Idelalisib group, Apelisib group and Copanlisib group. On days 3, 5 and 7, all rats in Na2SeO3, Idelalisib, Apelisib and Copanlisib groups were given subcutaneous injection into the nape with Na2SeO3 and control group was given the same amount of saline. For days 1-14, Idelalisib, Apelisib and Copanlisib were given by intragastric administration, respectively, and the same amount of saline was given to the control group and Na2SeO3 group. On the 15th day of the experiment, we selected the Idelalisib group with the best effect from all groups, separated their lenses, and further analyzed the crystal proteins, oxidative damage and apoptosis indexes.ResultsThe survival rate of rats in control and Idelalisib groups was 100%, the Na2SeO3 group was 37.5%, the Apelisib group was 25%, and the Copanlisib group was 0. According to the rat survival rate and lens score, we selected Idelalisib for further analysis of the crystallin, oxidative damage and apoptosis indexes. The results suggested that Na2SeO3 leads to cataract formation and crystallin precipitation. The levels of antioxidant enzymes GSH and SOD were decreased in the Na2SeO3 group, Nrf-2 and HO-1 were downregulated, Keap1 upregulated, and cleaved caspase-3 and Bax/Bcl-2 upregulated. Idelalisib significantly improved crystallin insolubility, reduced oxidative damage, and inhibited lens apoptosis.ConclusionIn summary, Idelalisib can significantly improve the progression of Na2SeO3-induced cataract in rats. In the future, it may be a potential effective drug candidate for the clinical treatment of cataract.

Polyphenol consumption and endurance exercise performance: A systematic review and meta-analysis of randomised controlled trials

Introduction: Consumption of polyphenols is proposed to reduce oxidative damage and increase nitric oxide synthesis and availability, thereby inducing physiological effects that enhance endurance exercise performance. Thus, the objective of this study was to systematically review the literature to evaluate the effects of consuming polyphenol-rich foods on endurance exercise performance. Methods: This study comprised a subanalysis of a larger systematic review. Electronic searches of Scopus, Web of Science, Ovid Medline, EMBASE and SportDiscus databases were performed. Studies were included if they involved consumption of foods containing polyphenols, and a measure of endurance exercise performance was reported. Risk of bias was assessed using the Cochrane RoB 2.0 and random effects meta-analysis was conducted, with subgroup analyses performed based on food type, sex, fitness level, and supplementation dosage and duration. Results: Data from 58 studies met eligibility criteria, with 55 studies included in the meta-analysis. Meta-analysis identified trivial but significant benefits for the overall consumption of polyphenol-rich foods on endurance exercise performance (SMD= 0.17, p<0.001). When evaluated by food type, grape, (nitrate-depleted) beetroot, French maritime pine, Montmorency cherry and pomegranate exhibited significant ergogenic effects, while no significant effects were evident for New Zealand blackcurrant, cocoa, ginseng, green tea or raisins. Significant positive effects were evident following both acute and multiple-day supplementation and meta-regression indicated that total polyphenols consumed did not moderate effect size. No effect of polyphenol consumption on exercise performance was found in females (SMD= 0.01, p=0.95), but small to moderate effects (SMD=0.21-0.76, p<0.001) were found in males across a range of fitness levels (V̇O2max 41-71 ml.kg.min-1). Limited evidence supported polyphenol-rich foods enhancing nitric oxide availability, improving vascular function, reducing oxidative damage or other physiological indicators that may explain these ergogenic benefits. Considerable heterogeneity in effect sizes of included studies was present, which may reflect variation in polyphenol bioavailability between foods, differences in study designs and subject characteristics. Discussion: Foods rich in polyphenols provide benefits for endurance exercise performance in males, but not females. These sex-specific benefits were observed across a range of fitness levels, from untrained to well-trained athletes. There is limited direct evidence of the mechanisms underpinning these effects. Other: The review protocol was registered on the Open Science Framework (https://osf.io/u7nsj). My co-authors and I acknowledge that we have no conflict of interest of relevance to the submission of this abstract.

Mechanism of reactive oxygen species generation and oxidative DNA damage induced by acrylohydroxamic acid, a putative metabolite of acrylamide

Acrylamide is formed during the heating of food and is also found in cigarette smoke. It is classified by the International Agency for Research on Cancer as a probable human carcinogen (Group 2A). Glycidamide, an epoxide metabolite of acrylamide, is implicated in the mechanism of acrylamide carcinogenicity. Acrylamide causes oxidative DNA damage in target organs. We sought to clarify the mechanism of acrylamide-induced oxidative DNA damage by investigating site-specific DNA damage and reactive oxygen species (ROS) generation by a putative metabolite of acrylamide, acrylohydroxamic acid (AA). Our results, using 32P-5′-end-labeled DNA fragments, indicated that, although AA alone did not damage DNA, AA treated with amidase induced DNA damage in the presence of Cu(II). DNA cleavage occurred preferentially at T and C, and particularly at T in 5′-TG-3′ sequences, and the DNA cleavage pattern was similar to that of hydroxylamine. The DNA damage was inhibited by methional, catalase, and Cu(I)-chelator bathocuproine, suggesting that H2O2 and Cu(I) are involved in the mechanism of DNA damage induced by AA treated with amidase. In addition, amidase-treated AA increased 8-oxo-7,8-dihydro-2′-deoxyguanosine formation in calf thymus DNA, an indicator of oxidative DNA damage, in a dose-dependent manner. In conclusion, hydroxylamine, possibly produced from AA treated with amidase, was autoxidized via the Cu(II)/Cu(I) redox cycle and H2O2 generation, suggesting that oxidative DNA damage induced by ROS plays an important role in acrylamide-related carcinogenesis.

Sinapic Acid Attenuates Rheumatoid Arthritis Through Reducing Inflammation and Oxidative Stress by Downregulating IκB Kinase.

Sinapic acid (SA) was reported to protect against inflammation in various types of diseases. However, the role of SA in rheumatoid arthritis remains unclear. This study was designed to investigate the role of SA on rheumatoid arthritis. Rheumatoid arthritis mouse model was established by collagen immunization [collagen-induced arthritis (CIA)]. Histological analysis of articular cartilage tissue was carried out by hematoxylin and eosin (H&E) staining. Serum concentrations of tumor necrosis factor alpha and interleukin 6 were determined through enzyme-linked immunosorbent assay (ELISA). Oxidative damage indexes such as superoxide dismutase activity, malondialdehyde detection, glutathione detection, and catalase were determined by biochemical analysis. The protein levels of related genes were determined using Western blot. In CIA model, SA treatment attenuated paw swelling and clinical score of arthritis, attenuated articular cartilage tissues edema and infiltration of inflammatory cells, suppressed inflammatory cytokines release, and attenuated oxidative damage indexes. Mechanically, SA suppressed immune responses through inhibiting the IκB kinase (IKKs). SA attenuates rheumatoid arthritis through reducing inflammation and oxidative stress by downregulating IKKs.

Malondialdehyde (MDA) level in cardiac tissue after induction of quail egg yolk (coturnix coturnix) in rats model (Rattus norvegicus)

Background and Aims: Oxidative stress plays an important role in the onset and progression of atherosclerosis and has impact on the development of cardiovascular diseases. Lipid peroxidation is one of the indicator of oxidative damage in cell membranes resulting from the ROS accumulation. Secondary oxidation products such as malondialdehyde is commonly measured to observe lipid peroxidation. Quail egg yolk (Coturnix coturnix) is known to have the highest cholesterol and lipid content compared to other avian species.

IN VITRO EVALUATION OF THE EFFECT OF GLUTATHIONE ON CASPASE SYSTEM AND OXIDATIVE DNA DAMAGE IN HIGH GLUCOSE CONDITION

This study was planned to investigate the effects of glutathione, known to be a strong antioxidant, on caspase-dependent apoptosis and oxidative DNA damage in the kidney cells (BHK-21) exposed to high glucose. BHK-21 cell line was cultivated by regular passages in vitro conditions. Study groups were set up as control, study and its combinations groups ((Glucose; (285 mM, HG), glutathione (250 µM)). After 24 hours of incubation, trypsinized cells were disrupted by freeze-thaw method and analyte was prepared. Caspase 3, 8, 9, M30 and 8-OHdG (oxidative DNA damage marker) levels were determined using commercial ELISA kits. M30, caspase 3, 8 and 9 which are the parameters of apoptosis, were found highest in the HG group. In GSH-treated groups, these parameters decreased slightly. There was no significant difference in the levels of 8-OHdG which is the indicator of oxidative DNA damage.

Bioaccumulation, Metabolism and the Toxic Effects of Chlorfenapyr in Zebrafish (Danio rerio)

Chlorfenapyr is widely used as an insecticide/miticide. Tralopyril, the active metabolite of chlorfenapyr, is used as an antifouling biocide in antifouling systems, and negatively affects aquatic environments. However, it is unclear whether tralopyril is a metabolite of chlorfenapyr in aquatic vertebrates, and there is little data on the bioaccumulation and toxicity of chlorfenapyr to aquatic vertebrates. In this study, the bioaccumulation and elimination of chlorfenapyr in zebrafish were assessed, and tralopyril, the active metabolite of chlorfenapyr, was determined. The effects of chronic exposure to chlorfenapyr on zebrafish liver and brain oxidative damage, apoptosis, immune response, and metabolome were investigated. These results showed that chlorfenapyr has a high bioaccumulation in zebrafish, with bioaccumulation factors of 864.6 and 1321.9 after exposure to 1.0 and 10 μg/L chlorfenapyr for 21 days, respectively. Chlorfenapyr at these concentrations also rapidly accumulated in zebrafish, reaching 615.5 and 10336 μg/kg on the second and third days of exposure, respectively. Chlorfenapyr was degraded to tralopyril in zebrafish; therefore, both chlorfenapyr and tralopyril should be considered when evaluating the risk of chlorfenapyr to aquatic organisms. In addition, chronic exposure caused oxidative damage, apoptosis, and immune disorders in zebrafish liver. Chronic exposure also altered the levels of endogenous metabolites in liver and brain. After 9 days of depuration, some indicators of oxidative damage, apoptosis, and immunity returned to normal levels, but the concentration of endogenous metabolites in zebrafish liver was still altered. Overall, these results provide useful information for evaluating the toxicity and environmental fate of chlorfenapyr in aquatic vertebrates.

Interactive Impacts of Beneficial Microbes and Si-Zn Nanocomposite on Growth and Productivity of Soybean Subjected to Water Deficit under Salt-Affected Soil Conditions.

Water stress or soil salinity is considered the major environmental factor affecting plant growth. When both challenges are present, the soil becomes infertile, limiting plant productivity. In this work a field experiment was conducted during the summer 2019 and 2020 seasons to evaluate whether plant growth-promoting microbes (PGPMs) and nanoparticles (Si-ZnNPs) have the potential to maintain soybean growth, productivity, and seed quality under different watering intervals (every 11 (IW0), 15 (IW1) and 19 (IW2) days) in salt-affected soil. The most extended watering intervals (IW1 and IW2) caused significant increases in Na+ content, and oxidative damage indicators (malondialdehyde (MDA) and electrolyte leakage (EL%)), which led to significant reductions in soybean relative water content (RWC), stomatal conductance, leaf K+, photosynthetic pigments, soluble protein. Subsequently reduced the vegetative growth (root length, nodules dry weight, and total leaves area) and seeds yield. However, there was an enhancement in the antioxidants defense system (enzymatic and non-enzymatic antioxidant). The individual application of PGPMs or Si-ZnNPs significantly improved leaf K+ content, photosynthetic pigments, RWC, stomatal conductance, total soluble sugars (TSS), CAT, POD, SOD, number of pods plant−1, and seed yield through decreasing the leaf Na+ content, MDA, and EL%. The combined application of PGPMs and Si-ZnNPs minimized the adverse impact of water stress and soil salinity by maximizing the root length, heavier nodules dry weight, leaves area, TSS and the activity of antioxidant enzymes, which resulted in higher soybean growth and productivity, which suggests their use under harsh growing conditions.

Oxidative stress and antioxidant defense in response to Dimethyl Phthalate in Chlorella vulgaris

Dimethyl phthalate (DMP) is a common organic pollutant that can accumulate in aquatic organisms and pose a threat to aquatic ecosystems. Many studies have shown that DMP is toxic to marine organisms, but the toxicity mechanism of DMP in microalgae is still unclear. We measured indicators of cell growth, oxidative damage and antioxidant enzyme activity in Chlorella vulgaris exposed to DMP. The degree of growth inhibition of C. vulgaris cells increased with the increase of DMP concentration. Hydrogen peroxide and malondialdehyde contents increased significantly in C.vulgaris exposed to 800 mg/L DMP. Glutathione peroxidase and Superoxide dismutase activities both decreased in C. vulgaris exposed to 100 mg/L DMP. These data indicated that DMP treatment altered the redox status of C. vulgaris and that oxidative damage may be the main mechanism of DMP toxicity. The results lay a foundation for understanding the impact of DMP on C. vulgaris stress and provide a theoretical basis for the protection of aquatic ecosystems.

Homeostatic alterations related to total antioxidant capacity, elemental concentrations and isotopic compositions in aqueous humor of glaucoma patients

Glaucoma is a multifactorial eye disease, characterized by progressive optic neurodegeneration. Elevation of the intraocular pressure is the main risk factor for glaucoma and is a consequence of an imbalance in the aqueous humor hydrodynamics, the physiology of which is influenced by the homeostatic equilibrium of essential elements, oxidative stress, and antioxidants. The aim of this work was to study local alterations in glaucomatous patients from two different, but connected, points of view: (i) the total antioxidant capacity (as an indicator of oxidative damage) and (ii) the concentration of mineral elements and their isotopic composition. Such objective was pursued using aqueous humor from patients diagnosed with pseudoexfoliation glaucoma (PEXG, n = 17) and primary open-angle glaucoma (POAG, n = 5) and age-matched control subjects (n = 16). The total antioxidant capacity (TAC) was examined in both aqueous humor and 60 serum samples (n = 20 controls, n = 20 for PEXG, and n = 20 for POAG), both showing higher TAC for the glaucoma population. The concentrations of the essential mineral elements (Cu, Fe, Mg, Na, P, and Zn) and the isotopic compositions of Cu and Zn were determined in aqueous humor using single-collector and multi-collector inductively coupled plasma–mass spectrometry, respectively. Significant differences were established for Mg and P levels when comparing the results for glaucomatous patients with those for the control population (p < 0.01 and p < 0.05 for Mg and P respectively, ANOVA and Kruskal-Wallis). The Zn isotopic composition was significantly shifted from that for the control population for PEXG patients. A significant difference in the isotopic composition of Zn was also established between the PEXG and POAG glaucoma cohorts.Graphical abstract

TRESK Regulates Gm11874 to Induce Apoptosis of Spinal Cord Neurons via ATP5i Mediated Oxidative Stress and DNA Damage.

Reportedly, TWIK-related spinal cord K+ (TRESK) deficiency in spinal cord neurons positively correlates with the mechanism underlying neuropathic pain (NP). However, the precise effects of TRESK on neurons of the spinal cord remain elusive. In the present study, we investigated the impact of TRESK silencing on spinal cord neurons to further elucidate the downstream mechanisms of TRESK. Herein, neurons of the dorsal spinal cord were cultured as a cell model for investigations. Apoptosis, oxidative stress, and DNA damage-related proteins were evaluated. Additionally, flow cytometry, microarray profiling, real-time polymerase chain reaction (PCR), western blotting, fluorescence in situ hybridization (FISH), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) were performed. In cultured neurons, the downregulation of TRESK mRNA expression induced apoptosis of dorsal spinal cord neurons. Using real-time PCR and western blotting, the upregulation of LncRNA Gm11874 (Gm11874) and ATP5i, screened from the gene chip, was confirmed. On silencing TRESK, expression levels of γ-H2AX, poly [ADP-ribose] polymerase 1 (PARP-1), FoxO1, FoxO3, MitoSOX, malondialdehyde (MDA), and 8-hydroxy-2' -deoxyguanosine (8-OHdG), which are known indices of oxidative stress and DNA damage, were significantly elevated. Moreover, ATP induced oxidative stress, DNA damage, and apoptosis were reduced by ATP5i siRNA. Finally, Gm11874 and ATP5i were co-expressed in spinal cord neurons in a FISH experiment, and the expression of ATP5i was positively regulated by Gm11874. These results implied that ATP5i induced oxidative stress and DNA damage, resulting in neuronal apoptosis, and Gm11874 was confirmed to act upstream of ATP5i. Our study revealed that TRESK silencing upregulated Gm11874 to induce apoptosis of spinal cord neurons, which resulted in ATP5i promoting oxidative stress and DNA damage. These findings could highlight the TRESK-mediated NP mechanism.

Inhibition of Epigenetic Reader BET Proteins by Apabetalone Counters Inflammation in Activated Innate Immune Cells from Fabry Disease Patients Receiving ERT

Abstract Fabry disease (FD) is a rare genetic disorder with a deficit in the degradation pathway of a glycolipid, globotriaocylceramide (Gb3). Gb3 deposits in various tissues evoke immune-mediated systemic inflammation, driving the progression of FD complications. In particular, cardiac and renal problems, leading causes of mortality in FD patients, are insufficiently managed by enzyme replacement therapy (ERT) in the long term. Here, we examined immune profiles in unstimulated peripheral blood mononuclear cells (PBMCs) from FD patients on ERT relative to untreated patients. The results showed an upregulation of the CCR2/MCP-1 axis, particularly in the presence of renal dysfunction. We also observed an increase of IL12B transcripts in unstimulated PBMCs when tracking the immune status over two years of ERT in patients with early-stage chronic kidney disease (CKD). Interestingly, pro-inflammatory responses in LPS-stimulated PBMCs were countered by apabetalone, a clinical-stage candidate that inhibits Bromodomain and Extra Terminal (BET) proteins, epigenetic readers. Apabetalone treatment dose dependently inhibited MCP-1 and IL-12 production by up to 90%. Apabetalone downregulated the transcription of other pro-inflammatory cytokines including TNF-α, IL-6 and IL-8 by 70%, 48% and 54%, respectively. Reactive oxygen species (ROS) are an indicator of oxidative damage caused by intracellular Gb3 deposits in FD patients. Apabetalone suppressed ROS levels by up to ~70% in LPS-stimulated neutrophils. Hence, apabetalone treatment may reduce pathological inflammation and oxidative stress in FD patients and thus complement ERT to optimize patient outcomes, warranting further investigation of apabetalone as a therapeutic for FD.

Chronic Exposure to Toluene and Heavy Metals and Changes in Indices of Liver Function, Inflammation and Oxidative DNA Damage among Automobile Workers

Background: Oxidative stress (OS), oxidative DNA damage and inflammatory response induced by chronic exposure to volatile organic compounds and heavy metals (HM) have been implicated in multiple organ dysfunction. The liver enzymes (alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT)), biomarkers of OS (nitric oxide (NO), glutathione (GSH), total antioxidant capacity (TAC), total plasma peroxides (TPP), malondialdehyde (MDA)) oxidative stress index (OSI)), oxidative DNA damage (8-hydroxy-2-deoxyguanosine (8-OHdG)), and inflammation marker (tumor necrosis factor alpha (TNF-I±)); heavy metals (cadmium (Cd), lead (Pb)) and urine hippuric acid (uHA) levels were assessed in automobile workers. Methods: Fifty automobile workers and 50 controls aged 18-60 years were enrolled into this study. The MDA, GSH, NO, TAC, TPP, ALT, ALP and GGT were estimated by colorimetry, 8-OHdG and TNF-I± by enzyme linked immunosorbent assay, Cd, Pb by atomic absorption spectrophotometry and uHA by high performance liquid chromatography. Data were analyzed using t-test and correlation analysis at p <0.05. Results: Automobile workers had significantly higher liver enzymes, lipid peroxidation, oxidative stress, oxidative DNA damage, nitric oxide, HM, uHA and lower total antioxidants relative to controls. Heavy metals were positively associated with MDA, TPP and OSI; TPP with duration of exposure; ALP with number of working hours; and liver enzymes with OSI only in automobile workers. Conclusion: Association of exposure to toluene and heavy metals with increased liver enzymes activity, lipid peroxidation, oxidative stress, oxidative DNA damage, and depressed antioxidants in automobile workers suggest increased risk of hepatotoxicity and hepatocellular carcinogenesis.

Assessment spermatogenic cell apoptosis and the transcript levels of metallothionein and p53 in Meretrix meretrix induced by cadmium

Cadmium (Cd) has been widely used in industry and can accumulate in the water, soil, and food. Meretrix meretrix is one of the marine shellfishes cultivated for economic purpose in China. The increasing Cd levels in coastal marine water could adversely affect the economic benefits of shellfish cultivation. In the present study, M. meretrix were exposed to different Cd2+ concentrations (0, 1.5, 3, 6, and 12 mg L-1) for 5 d to evaluate the effects of Cd on spermatogenic cell. The Cd accumulation, survival rate and the indices of oxidative stress and apoptosis were determined in the spermatogenic cells of M. meretrix. The expression levels of p53 and metallothionein (MT) mRNA were also measured in the spermatogenic cells. Cd accumulation and the mortality rate of spermatogenic cells were found to increase in a dose-response manner with Cd2+ concentrations. Histopathology changes, especially the damage of membranous structure, were more severe as the Cd2+ levels in the testis became higher. The indexes of oxidative stress, including reactive oxygen species, malondialdehyde, protein carbonyl derivates and DNA-protein crosslinks all increased after exposure to Cd2+. However, the total antioxidant capacity gradually decreased with the increasing Cd2+ concentration. In addition, exposure to Cd2+ increased the apoptotic rate and caspase-3 and 9 activities but decreased the level of mitochondrial membrane potential and cytochrome C oxidase in the spermatogenic cells. MT mRNA expression increased in lower Cd2+ concentration treated groups whereas decreased in higher groups, while the p53 mRNA expression increased in a dose-response manner with Cd2+ and was positively correlated with the oxidative damage indices. These results indicated that Cd2+ caused oxidative stress and p53 induced apoptosis in the spermatogenic cells, and thus decreased the survival rate of sperm cells. This finding highlights that Cd can reduce the reproductive capacity of M. meretrix, thus threatening to wild shellfish populations and reducing the efficiency of shellfish farming.