Inhibition Of Superoxide Dismutase Activity Research Articles

A self-assembled theranostic nanoplatform for efficient tumor sonodynamic-gas therapy

The powerful antioxidant system in tumor tissues limits the efficacy of sonodynamic therapy (SDT) by reducing the availability of the produced reactive oxygen species (ROS). To address this problem, a pH/ultrasound (US) responsive theranostic nanoplatform is elaborately constructed by self-assembly of nitrogen-doped graphene quantum dots (NGQDs), benzothiazole sulfinate (BTS), Gd3+ ions and poly(lactide-co-glycolide)-b-poly(ethylene glycol) (PLGA-PEG). The NGQD component with excellent sonodynamic activity wrapped in the resulting Gd-NGQDs/BTS@PLGA-PEG can efficaciously respond to ultrasonic irradiation and cause abundant ROS generation. Importantly, in response to acidic tumor microenvironment, the loaded BTS sustainably releases SO2, which induces the increase of intracellular •O2− level by inhibiting superoxide dismutase activity and further promotes the enrichment of ROS by reducing intracellular glutathione level and suppressing glutathione peroxidase 4 activity. The well-designed SO2 gas therapy-enhanced SDT strategy leads to high oxidative stress in tumor cells and effective inhibition of the growth of bilateral tumors with only a single intravenous injection of the nanomedicine and a subsequent single US irradiation. Gd-NGQDs/BTS@PLGA-PEG can also be employed as a good T1-weighted magnetic resonance imaging contrast agent to guide tumor therapy. This innovative therapeutic strategy developed in our contribution may provide a new paradigm for the exploration of deep tumor therapy.

Nitrate reduces copper toxicity by preventing oxidative stress and inhibiting copper translocation from roots to shoots in Liriodendron Chinense.

Nitrogen forms can affect metal accumulation in plants and tolerance to metals, but a few published studies on the effects on Cu toxicity and Cu accumulation in plants are scarce. Thus, the objective of this study was to evaluate the responses of Liriodendron chinense to different nitrogen forms, by the oxidative stress, antioxidant enzymes system, GSH-AsA cycle, Cu uptake, translocation, and accumulation under Cu stress. We found that Cu-induced growth inhibiting was alleviated by added exclusive NO3--N. Adding N as NH4+-N with or without NO3--N was aggravated as evidenced by significantly elevated malonaldehyde (MDA) and hydrogen peroxide (H2O2) compared to N-Null. Cu exposure and adding NH4+-N inhibited superoxide dismutase activity, but remarkably stimulated the activities of catalase and peroxidase, the efficiency of glutathione-ascorbate (GSH-AsA) cycle, and the activity of glutathione reductase and nitrate reductase, with respect to the control. However, adding exclusive NO3--N progressively restored the alteration of antioxidant to prevent Cu-induced oxidative stress. Additionally, adding exclusive NO3--N significantly promoted the Cu uptake and accumulation in roots, but reduced Cu concentration in leaves, accompanied by the inhibited Cu translocation factor from roots to shoots by 36.7%, when compared with N-Null. Overall, adding NO3--N alleviated its Cu toxicity by preventing Cu-induced oxidative stress and inhibiting Cu translocation from roots to shoots, which provides an effective strategy for phytostabilization in Cu-contaminated lands.

The safety evaluation of Shenze Shugan capsule and mechanism of apoptosis induced by five potentially nephrotoxic components

Ethnopharmacological relevanceShenze Shugan capsule is a prescription of traditional Chinese medicine for nonalcoholic steatohepatitis treatment. It includes Rhei Radix et Rhizoma (RR), Cassiae Semen (CS) and Alismatis Rhizoma(AR), which widely contains rhein, emodin, aurantio-obtusin, alisol A and alisol B 23-monoacetate. Aim of the studyIn this study, we aimed to explore the safety of the medicine, and further elucidate the mechanism of apoptosis induction in HK-2 cells by five components, including rhein, emodin, aurantio-obtusin, alisol A and alisol B 23-monoacetate. Materials and methodsWe investigated the nephrotoxicity of Shenze Shugan capsule, including RR, CS, AR and mixed herbs given for two months in rats. Superoxide dismutase (SOD) in kidney tissues, urea nitrogen (BUN) and creatinine (CRE) in serum were detected, and renal pathology analysis was performed. In cell experiments, the apoptotic rate and cell cycle distribution of HK-2 cells were tested by flow cytometry. The levels of mitochondrial membrane potential (ΔΨm) and related protein expression in mitochondrial pathway were measured as well. ResultsWe confirmed that two months of administering high doses(60 times the dose for clinical use in adults) of RR, CS or mixed herbs upregulated the levels of CRE and RUN, inhibited SOD activity, and increased the degree of tubular degeneration and glomerular dilatation, but Shenze Shugan capsule has no significant differences in renal structure or renal function. In addition, we found that five components all concentration-dependently inhibited HK-2 cells proliferation and induced apoptosis, especially aurantio-obtusin as the novel nephrotoxic component. Rhein and emodin significantly induced S/M accumulation, but aurantio-obtusin, alisol A and alisol B 23-monoacetate significantly induced G1/M accumulation in HK-2 cells. Similarly, they could induce Caspase3 activation, loss of mitochondrial membrane potential (ΔΨm), and down-regulation of Bcl-2 and up-regulation of Bax. ConclusionsThrough a two-month subchronic toxicity study in rats, our preliminary determination is that this formulation is safe and reliable for long-term use. Interestingly, the potentially toxic herbs such as RR, CS, AR can reduce toxicity by drug compatibility. When further exploring the mechanism of action of toxic herbs, we found that mitochondrial pathway is involved in the apoptosis of HK -2 cells induced by rhein, emodin, aurantio-obtusin, alisol A and alisol B 23-monoacetate. Our findings provide new ideas for safety studies of Shenze Shugan capsule.

Mechanistic Aspects of Biphenyl Urea-Based Analogues in Triple-Negative Breast Cancer Cell Lines.

Triple-negative breast cancer (TNBC) poses significant challenges due to its aggressive nature and limited treatment options. In this study, we investigated the impact of urea-based compounds on TNBC cells to uncover their mechanisms of action and therapeutic potential. Notably, polypharmacology urea analogues were found to work via p53-related pathways, and their cytotoxic effects were amplified by the modulation of oxidative phosphorylation pathways in the mitochondria of cancer cells. Specifically, compound 1 demonstrated an uncoupling effect on adenosine triphosphate (ATP) synthesis, leading to a time- and concentration-dependent shift toward glycolysis-based ATP production in MDA-MB-231 cells. At the same time, no significant changes in ATP synthesis were observed in noncancerous MCF10A cells. Moreover, the unique combination of mitochondrial- and p53-related effects leads to a higher cytotoxicity of urea analogues in cancer cells. Notably, the majority of tested clinical agents, but sorafenib, showed significantly higher toxicity in MCF10A cells. To test our hypothesis of sensitizing cancer cells to the treatment via modulation of mitochondrial health, we explored the combinatorial effects of urea-based analogues with established chemotherapeutic agents commonly used in TNBC treatment. Synergistic effects were evident in most tested combinations in TNBC cell lines, while noncancerous MCF10A cells exhibited higher resistance to these combination treatments. The combination of compound 1 with SN38 displayed nearly 60-fold selectivity toward TNBC cells over MCF10A cells. Encouragingly, combinations involving compound 1 restored the sensitivity of TNBC cells to cisplatin. In conclusion, our study provides valuable insights into the mechanisms of action of urea-based compounds in TNBC cells. The observed induction of mitochondrial membrane depolarization, inhibition of superoxide dismutase activity, disruption of ATP synthesis, and cell-line-specific responses contribute to their cytotoxic effects. Additionally, we demonstrated the synergistic potential of compound 1 to enhance the efficacy of existing TNBC treatments. However, the therapeutic potential and underlying molecular mechanisms of urea-based analogues in TNBC cell lines require further exploration.

Combined application of biochemical and chemical parameters in the liver of round goby Neogobius melanostomus (Pallas, 1814) for assessing the quality of water bodies

Bioindication assessment of three areas in the Sea of Azov (Pre-strait (PS), Open Sea (OS) and Taganrog Bay (TB)) by using some biochemical (superoxide dismutase (SOD), catalase (CAT), aminotransferase, cholinesterase (ChE) activities, albumin (Alb) content, levels of oxidized proteins (OP) and lipid peroxidation (LPO) as well as chemical (mercury (Hg), lead (Pb), cadmium (Cd), arsenic (As), iron (Fe), copper (Cu), zinc (Zn)) parameters in the liver of round goby Neogobius melanostomus (Pallas, 1814) was carried out. The results of the CE analysis in the liver of fish from three locations did not exceed the maximum permissible concentrations (MPC), with the exception of Cu. Cu content amounted 1.4, 1.7 and 9.5 MPC in the liver of fish from PS, OS and TB regions respectively. Concentrations of Hg, Pb, Fe and Cu were higher in the liver of specimens from TB and decreased in a number of locations: TB → OS → PS, that is in line with the data on biochemical analyses. Inhibition of SOD activity, imbalance in the processes of peroxide neutralization as well as the shift of prooxidant–antioxidant reactions towards LPO in the liver of gobies from TB region indicate toxic chemical impacts and less favorable ecological state of this location compared to PS and OS areas. High informativeness of the combined use of the biochemical and chemical parameters in the liver of round goby for assessing the quality of marine environment with different levels of chemical pollution was shown.

Microplastics Pollution and Their Potential Impact in Marine Systems: A Case Study in Shandong Peninsula, China

Shandong Province is among China’s major marine provinces, and the marine environment in Shandong supports important economic activities, such as fishing, aquaculture, and tourism. The presence of microplastics in the marine environment can impact these activities and the livelihoods of people who depend on them. Therefore, studying microplastics in Shandong Peninsula is crucial for understanding and mitigating these impacts. In this research, the microplastics were detected with abundance ranging from 50.67 to 315.2 particles/kg dry weight, and polyethylene and polypropylene have dominant proportions in beach sand and sediments. Microplastics were also detected in holothurians and oysters with detection rates at 16.7% and 38.9%, respectively. This study highlighted that microplastics pollution in the Shandong Peninsula was closely related to anthropogenic activities. Moreover, microplastics have inhibited superoxide dismutase activity and significantly increased malondialdehyde during zebrafish growth, which provided a potential effect between microplastics and marine organism. Human activities contribute to microplastics pollution in the environment, while at the same time, the presence of microplastics can impact marine organism and human activities. To mitigate the impacts of microplastics on the Shandong Peninsula, it is important to reduce the sources of microplastics and to develop effective management strategies for controlling their spread and impact.

Cadmium (Cd) exposure through Hyphantria cunea pupae reduces the parasitic fitness of Chouioia cunea: A potential risk to its biocontrol efficiency

Heavy metal contamination has been regarded as an environmental variable that affects the efficiency of pest biological control, but the parasitic fitness of parasitoids under heavy metal stress is poorly understood. Herein, the effect of Cd exposure through the host pupa of Hyphantria cunea on the parasitic fitness of Chouioia cunea was investigated, and the mechanism by which Cd exposure affects the interaction between H. cunea and C. cunea from the perspective of innate immunity in host insect and the oxidative status in the parasitoid offspring was explored. Our results indicated that Cd can be transferred from the H. cunea pupae to the parasitoid offspring, and the transfer coefficient reflected biological amplification. There were no significant differences in the rates of parasitism success and offspring emergence between the untreated and Cd-treated groups. However, after parasitizing Cd-accumulated pupae, the parasitic fitness of offspring wasps (e.g., the number, individual size and life span) decreased significantly. Under Cd exposure, the cellular and humoral immunity of H. cunea pupae decreased significantly. Compared with the untreated group, the H2O2 content of parasitoid offspring in the Cd-treated group was significantly increased. Cd exposure significantly inhibited superoxide dismutase activity in parasitoid offspring, but the contents of ascorbic acid and glutathione were significantly increased by Cd stress. Taken together, these results indicate that Cd exposure reduces the cyclic utilization efficiency of C. cunea on H. cunea pupae. The oxidative status of parasitoid offspring triggered by Cd exposure could be responsible for the reduced parasitic fitness of C. cunea on Cd-accumulated H. cunea pupae.

Chenodeoxycholic Acid-Amikacin Combination Enhances Eradication of Staphylococcus aureus.

The rise of antibiotic resistance and dearth of novel antibiotics have posed a serious health crisis worldwide. In this study, we screened a combination of antibiotics and nonantibiotics providing a viable strategy to solve this issue by broadening the antimicrobial spectrum. We found that chenodeoxycholic acid (CDCA), a cholic acid derivative of the traditional Chinese medicine (TCM) Tanreqing (TRQ), synergizes with amikacin against Staphylococcus aureus in vitro, and this synergistic killing was effective against diverse methicillin-resistant S. aureus (MRSA) variants, including small-colony variants (SCVs), biofilm strains, and persisters. The CDCA-amikacin combination protects a mouse model from S. aureus infections. Mechanistically, CDCA increases the uptake of aminoglycosides in a proton motive force-dependent manner by dissipating the chemical potential and potentiates reactive oxygen species (ROS) generation by inhibiting superoxide dismutase activity. This work highlights the potential use of TCM components in treating S. aureus-associated infections and extend the use of aminoglycosides in eradicating Gram-positive pathogens. IMPORTANCE Multidrug resistance (MDR) is spreading globally with increasing speed. The search for new antibiotics is one of the key strategies in the fight against MDR. Antibiotic resistance breakers that may or may not have direct antibacterial action and can either be coadministered or conjugated with other antibiotics are being studied. To better expand the antibacterial spectrum of certain antibiotics, we identified one component from a traditional Chinese medicine, Tanreqing (TRQ), that increased the activity of aminoglycosides. We found that this so-called agent, chenodeoxycholic acid (CDCA), sensitizes Staphylococcus aureus to aminoglycoside killing and protects a mouse model from S. aureus infections. CDCA increases the uptake of aminoglycosides in a proton motive force-dependent manner by dissipating the chemical potential and potentiates ROS generation by inhibiting superoxide dismutase activity in S. aureus. Our work highlights the potential use of TCM or its effective components, such as CDCA, in treating antibiotic resistance-associated infections.

Aerobic Treadmill Exercise Upregulates Epidermal Growth Factor Levels and Improves Learning and Memory in d-galactose-Induced Aging in a Mouse Model.

Previous studies have demonstrated that exercise improves cognitive function in Alzheimer's disease mice but the exact mechanism needs further studies. This research aimed to study the effects of aerobic treadmill exercise on epidermal growth factor (EGF) levels and learning and memory in d-galactose-induced aging in a mouse model. Forty male Kunming mice were analyzed in this study and randomly divided into 4 groups: control (C group), aerobic exercise (AE group), d-galactose (D-gal group), and d-galactose + aerobic exercise (D-gal + AE group). The C and AE groups received a daily mid-scapular subcutaneous injection of .9% saline for 40days. Mice in the D-gal and D-gal + AE groups were subcutaneously injected with d-galactose (1.25mg/kg) once daily for 40days. The mice in the AE group and D-gal + AE group completed 40days of aerobic treadmill exercise. Learning and memory were evaluated by step-down tests. Specifically, 24h after the behavioral test, blood was collected and brain tissue was extracted, and superoxide dismutase (SOD) and acetylcholinesterase activities were detected. The neurons in the CA1 and CA3 regions of the hippocampus were counted by Nissl staining. The number of EGF-positive cells was observed by immunohistochemical methods. In the learning test, the reaction time in the D-gal group increased significantly (P < .05), while the error numbers in the D-gal group tended to decrease compared with AE, D-gal + AE, and C groups. In the memory test, the latency of mice in the D-gal group was lower, while the error in this group was higher than in the other groups (P < .05). The activities of SOD and acetylcholinesterase were lower in the D-gal group than in the other groups (P < .05). The number of EGF-positive cells and neurons in the hippocampal CA1 and CA3 regions in the D-gal + AE group was higher compared to those in the D-gal group (P < .05), and lower in groups with mice that were not injected with d-galactose. Aerobic treadmill exercise inhibited SOD activity, increased EGF-positive cells, and decreased neuronal death and apoptosis, thereby improving learning and memory in the mouse model of d-galactose-induced aging.

Mitochondrial dysfunction due to in vitro exposure to atrazine and its metabolite in striatum.

Atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) has beendescribed as a potential toxic for dopaminergic metabolism both in vivo and in vitro. Its main metabolite diamino-chloro triazine (DACT) has been shown to achieve higher levels in brain tissue than atrazine. The aim of this study was to evaluate the in vitro effects of atrazine and DACT on striatal mitochondrial function, active oxygen species generation, and nitric oxide (NO) content. Incubation of mitochondria with atrazine (10 µM) was not able to modify oxygen consumption. However, a 50% increase in malate-glutamate state 4 respiratory rates was observed after DACT treatment (100 µM) without changes in respiratory state 3. Atrazine was able to inhibit complex I-III activity by 30% and DACT induced a tendency to decrease by 17% in the striatum. Regardingreactive oxygen species(ROS), DACT increased H2 O2 production by 43%. Also, superoxide anion levels were higher (14%) after atrazine exposure than in control mitochondria. Incubation of striatal mitochondria with atrazine and DACT induced membrane depolarization by 15% and 19%, respectively. Also, atrazine increased NO content by 10% but no significant changes were observed after exposure of mitochondria to DACT. Glutathione peroxidase activity was inhibited (56%) by DACT and atrazine inhibited superoxide dismutase activity by 60%. Also, cardiolipin oxidation (15%) was observed after atrazine treatment. Summing up, the obtained results suggest that in vitro atrazine and DACT induce ROS production affecting striatal mitochondrial function. The atrazine effects would be attributed to a direct effect on the mitochondrialrespiratory chain and superoxide dismutase activity while DACT appears to disturb glutathione-related enzyme system.

Response pathways of superoxide dismutase and catalase under the regulation of triclocarban-triggered oxidative stress in Eisenia foetida: Comprehensive mechanism analysis based on cytotoxicity and binding model

Triclocarban (TCC) is an emerging environmental contaminant, posing potential ecological risks. Displaying a high accumulation effect and 120-day half-life in the soil environment, the toxic effects of TCC to soil organisms have been widely reported. Previous studies have confirmed that TCC can induce the oxidative stress and changes in superoxide dismutase (SOD) and catalase (CAT) activities in earthworms, but the underlying mechanisms of oxidative stress and disorder in antioxidant enzyme activities induced by TCC have not yet been elucidated. Here, we explored the multiple response mechanisms of SOD and CAT under the regulation of oxidative stress induced by TCC. Results indicated that higher-dose (0–2.0 mg/L) TCC exposure triggered the overproduction of ROS in Eisenia foetida coelomocytes, causing oxidative damage and a decrease in cell viability that was response to ROS accumulation. The TCC-induced inhibition of intracellular SOD/CAT activity was found under the regulation of oxidative stress (SOD: 29.2 %; CAT: 18.5 %), and this effect was blunted by antioxidant melatonin. At the same time, the interaction between antioxidative enzymes and TCC driven by various forces (SOD: electrostatic interactions; CAT: van der Waals forces and hydrogen bonding) led to inhibited SOD activity (9.84 %) and enhanced CAT activity (17.5 %). Then, to elucidate the binding mode of TCC, we explored the changes in SOD and CAT structure (protein backbone and secondary structure), the microenvironment of aromatic amino acids, and aggregation behavior through multispectral techniques. Molecular docking results showed that TCC inhibited SOD activity in a substrate competitive manner and enhanced CAT activity by the stabilizing effects of TCC on the heme groups. Collectively, this study reveals the response mechanisms of SOD/CAT under the regulation of TCC-triggered oxidative stress and shed a new light on revealing the toxic pathways of exogenous pollutants on antioxidant-related proteins function.

Copper Chaperone for Superoxide Dismutase FoCCS1 in Frankliniella occidentalis May Be Associated with Feeding Adaptation after Host Shifting.

Simple SummaryWestern flower thrips (Frankliniella occidentalis) have a wide range of hosts. Therefore, they can colonize new host plants with each seasonal change. This study examined whether the superoxide dismutase (SOD) gene regulates the feeding adaptation of F. occidentalis after host shifting. The coding sequences for CCS1 and MnSOD2 in F. occidentalis were cloned and the corresponding amino acid sequence was predicted, and the mRNA expression levels of these two genes at different developmental stages were determined. Further, the mRNA expression levels of FoCCS1 and FoMnSOD2 in second-instar larvae and adult females transferred to kidney bean and broad bean plants for rearing were analyzed. Decreasing the mRNA levels of FoCCS1 and SOD activity by RNA interference significantly reduced the survival rate and fecundity of adult F. occidentalis females. These findings provide a reference for analyzing the adaptive mechanism of F. occidentalis after host shifting.Western flower thrips (Frankliniella occidentalis) pose a serious threat to the global vegetable and flower crop production. The regulatory mechanism for superoxide dismutase (SOD) in the feeding adaptation of F. occidentalis after host shifting remains unclear. In this study, the copper chaperone for SOD (CCS) and manganese SOD (MnSOD) genes in F. occidentalis were cloned, and their expression levels at different developmental stages was determined. The mRNA expression of FoCCS1 and FoMnSOD2 in F. occidentalis second-instar larvae and adult females of F1, F2, and F3 generations was analyzed after shifting the thrips to kidney bean and broad bean plants, respectively. The F2 and F3 second-instar larvae and F2 adult females showed significantly upregulated FoCCS1 mRNA expression after shifting to kidney bean plants. The F1 second-instar larvae and F2 adult females showed significantly upregulated FoCCS1 mRNA expression after shifting to broad bean plants. The RNA interference significantly downregulated the FoCCS1 mRNA expression levels and adult females showed significantly inhibited SOD activity after shifting to kidney bean and broad bean plants. F. occidentalis adult females subjected to RNA interference and released on kidney bean and broad bean leaves for rearing, respectively, significantly reduced the survival rate and fecundity. These findings suggest that FoCCS1 plays an active role in regulating the feeding adaptation ability of F. occidentalis after host shifting.

The repositioned drugs disulfiram/diethyldithiocarbamate combined to benznidazole: Searching for Chagas disease selective therapy, preventing toxicity and drug resistance.

Chagas disease (CD) affects at least 6 million people in 21 South American countries besides several thousand in other nations all over the world. It is estimated that at least 14,000 people die every year of CD. Since vaccines are not available, chemotherapy remains of pivotal relevance. About 30% of the treated patients cannot complete the therapy because of severe adverse reactions. Thus, the search for novel drugs is required. Here we tested the benznidazole (BZ) combination with the repositioned drug disulfiram (DSF) and its derivative diethyldithiocarbamate (DETC) upon Trypanosoma cruzi in vitro and in vivo. DETC-BZ combination was synergistic diminishing epimastigote proliferation and enhancing selective indexes up to over 10-fold. DETC was effective upon amastigotes of the BZ- partially resistant Y and the BZ-resistant Colombiana strains. The combination reduced proliferation even using low concentrations (e.g., 2.5 µM). Scanning electron microscopy revealed membrane discontinuities and cell body volume reduction. Transmission electron microscopy revealed remarkable enlargement of endoplasmic reticulum cisternae besides, dilated mitochondria with decreased electron density and disorganized kinetoplast DNA. At advanced stages, the cytoplasm vacuolation apparently impaired compartmentation. The fluorescent probe H2-DCFDA indicates the increased production of reactive oxygen species associated with enhanced lipid peroxidation in parasites incubated with DETC. The biochemical measurement indicates the downmodulation of thiol expression. DETC inhibited superoxide dismutase activity on parasites was more pronounced than in infected mice. In order to approach the DETC effects on intracellular infection, peritoneal macrophages were infected with Colombiana trypomastigotes. DETC addition diminished parasite numbers and the DETC-BZ combination was effective, despite the low concentrations used. In the murine infection, the combination significantly enhanced animal survival, decreasing parasitemia over BZ. Histopathology revealed that low doses of BZ-treated animals presented myocardial amastigote, not observed in combination-treated animals. The picrosirius collagen staining showed reduced myocardial fibrosis. Aminotransferase de aspartate, Aminotransferase de alanine, Creatine kinase, and urea plasma levels demonstrated that the combination was non-toxic. As DSF and DETC can reduce the toxicity of other drugs and resistance phenotypes, such a combination may be safe and effective.

Albiflorin ameliorates inflammation and oxidative stress by regulating the NF-κB/NLRP3 pathway in Methotrexate-induced enteritis

Methotrexate (MTX) treats various diseases but also damages intestinal barrier and leads to enteritis. Albiflorin (ALB) has a variety of pharmacological effects, including antioxidant, anti-inflammation and anti-apoptosis. In the present study, we evaluated the therapeutic effect of ALB on MTX-induced enteritis and investigated the possible mechanisms involved. Male SD rats were intraperitoneally injected with 7 mg/kg MTX for three consecutive days to establish the enteritis model. ALB (20 or 40 mg/kg/day) was intragastrically administrated since two days prior MTX treatment and lasted for six days. We found that ALB treatment increased body weight and intestinal weight of rats with MTX injection. The disease activity index (DAI) score was also decreased after ALB administration. In histological examination, ALB treatment attenuated inflammatory cells infiltration and promoted survival of goblet cells. In detection of inflammatory-associated factors, ALB treatment decreased CD68+ cells infiltration, inhibited myeloperoxidase activity, and suppressed intercellular cell adhesion molecule-1 and cyclooxygenase-2 expression. Additionally, ALB reduced malondialdehyde, glutathione levels, inhibited superoxide dismutase activity and suppressed reactive oxygen species production. Moreover, ALB treatment effectively inhibited NLRP3, as well as caspase 1 p20 and interleukin (IL)-1β and 18 expression. Finally, nuclear factor-κB (NF-κB) p65 phosphorylation and nuclear translocation were also demonstrated to be blocked upon ALB treatment. In conclusion, our findings indicated that ALB alleviated MTX-induced enteritis via inhibiting the NF-κB/NLRP3 pathway.

Free radical processes in rat liver in experimental diabetes, malignant growth and their combination

Purpose of the study. To study the intensity of lipid peroxidation (LPO) and indicators of antioxidant protection in the liver of rats of different sexes with combined pathology – the growth of Guerin's carcinoma in presence of experimental type 1 diabetes mellitus.Materials and methods. The levels of malondialdehyde (MDA) and diene conjugates (DC), the activities of superoxide dismutase (SOD) and catalase, and the levels of vitamins A and D were studied by conventional spectrophotometric methods in liver tissue samples from males and females with alloxan‑induced diabetes mellitus (DM), transplanted Guerin’s carcinoma (GC) and in animals with combined pathologies (GC growth in presence of diabetes mellitus), as well as in intact rats.Results. The levels of both LPO markers in intact females were significantly higher than in males: MDA by 3 times, DC by 2 times. The development of DM led to an increase in the content of MDA and DC in the liver of males compared with intact animals, which indicated an intensification of LPO. At the same time, there was an almost twofold increase in SOD activity, a decrease in catalase activity, and an increase in the ratio of enzymes by 2.8 times. In females, there was no increase in LPO in the liver; there was only a less pronounced increase in SOD activity than in males. At the same time, SOD activity in males was 37 % higher than in females, and catalase activity, on the contrary, was lower by 33 %, which caused a twofold excess of the ratio of enzymes in males relative to females. Among animals with GC, only males had an increase in SOD by 2.5 times, a decrease in catalase activity, and an increase in the SOD/Catalase ratio by 3 times relative to intact animals, which may be due to pronounced sex differences the SOD activity and SOD/Catalase ratio in males were 3–4 times higher than that of females. The levels of MDA and DC in males with GC growing in presence of DM remained elevated, although to a somewhat lesser extent than in DM alone, while in females both indicators were reduced. The ratio of vitamins E and A increased only in males. SOD activity and SOD/Catalase coefficient in animals of both sexes with combined pathologies were significantly lower than in groups with only DM or GC.Conclusions. The data showed that diabetes mellitus changes the specificity of oncogenesis depending on the sex of the animals. Males with the growth of Guerin's carcinoma in presence of diabetes in conditions of inhibition of SOD activity and a decrease in the ratio of conjugated enzymes of the first line of antioxidant defense demonstrate an intensification of LPO accompanied by an increase in the non‑enzymatic unit of the antioxidant system, while there is no increase in LPO in females, and the indicators of antioxidant activity change to a lesser extent.

Investigation of nickel sulfate-induced cytotoxicity and underlying toxicological mechanisms in human umbilical vein endothelial cells through oxidative stress, inflammation, apoptosis, and MAPK signaling pathways.

Growing evidence indicates that nickle and its compounds have adverse effects on the cardiovascular system. In this study, the cytotoxic insults caused by nickel sulfate (NiSO4 ) in human umbilical vein endothelial cells (HUVECs) were explored by examining cell viability, oxidative stress, inflammation, apoptosis, and MAPK signaling pathway activity. Cultured HUVECs were treated with varying concentrations of NiSO4 (0, 62.5, 250, and 1000 μM) for 24 h. Subsequently, markers of oxidative stress, inflammation, apoptosis, and MAPK signaling pathways were analyzed using biochemical assays, real-time quantitative polymerase chain reaction, and western blot. Rates of apoptosis were evaluated using flow cytometry. The results showed that NiSO4 exerted dose- and time-dependent inhibitory effects on cell growth. It induced oxidative stress and lipid peroxidation by increasing the generation of reactive oxygen species, the oxidized glutathione to reduced glutathione ratio (GSSG/GSH ratio), and malondialdehyde levels. Further, it inhibited superoxide dismutase activity in HUVECs. Flow cytometry analysis results revealed that NiSO4 (62.5-1000 μM) could induce apoptosis in HUVECs. The protein and gene expressions of cleaved Caspase 3 and Bax were elevated, and those of Bcl-2 and Bcl-XL were reduced after NiSO4 treatment. Additionally, NiSO4 triggered inflammation in HUVECs, increasing the protein and mRNA levels of IL-6 and TNF-α and reducing those of TGF-β. Furthermore, western blot findings revealed that NiSO4 could activate MAPK signaling pathways, upregulating p38, JNK, and ERK1/2 in HUVECs by increasing the levels of p-P38，p-JNK, and p-ERK1/2 in a dose-dependent manner. MAPK pathway inhibitors (10μM SB203580 and 10μM SP600125) could attenuate the NiSO4 -induced increase in apoptosis and inflammation in HUVECs. They could also attenuate the dysregulation of inflammatory factors and related proteins caused by high-dose NiSO4 exposure. Interestingly, while the MEK inhibitor U0126 (10μM) enhanced NiSO4 -induced apoptosis in HUVECs, it reduced cell inflammation. Taken together, these experimental results suggest that NiSO4 can inhibit cell growth, induce oxidative stress, and trigger subsequent inflammatory responses and apoptosis in HUVECs. These effects may be mediated by the P38 and JNK MAPK stress response pathways.

Disrupting circ-GNB4 mitigates high glucose-induced human mesangial cells injury by regulating the proliferation, ECM accumulation, inflammation and oxidative stress through circ-GNB4/miR-23c/EGR1 pathway [RETRACTED

Diabetic nephropathy (DN) is the most common complication of diabetes mellitus. Although G protein subunit beta 4 (GNB4)-derived circular RNA (circ-GNB4; hsa_circ_0068087) is a promising candidate biomarker in diabetes mellitus, whether circ-GNB4 participates in DN occurrence and development remains unknown. Herein, we focused on DN-associated human renal mesangial cells (HRMCs) injury, and HRMCs were exposed in high glucose (HG) condition. Using quantitative polymerase chain reaction and western blotting, we found that circ-GNB4 and early growth response factor 1 (EGR1) were upregulated, whereas microRNA (miR)-23c was downregulated in DN patients' sera and HG-stimulated HRMCs. HG-induced injuries were measured by MTS method, western blotting, enzyme-linked immunosorbent assay and other special assay kits. Consequently, HG could inhibit superoxide dismutase activity, but induce cell proliferation and levels of malondialdehyde, Fibronectin, Collagen I, Collagen IV, interleukin-6, interleukin-1β, and tumor necrosis factor-α. However, HG-induced these injuries were overall suppressed by silencing circ-GNB4 or overexpressing miR-23c. Moreover, miR-23c knockdown could counteract the effect of circ-GNB4 deficiency, and EGR1 restoration abrogated miR-23c overexpression role in HG-stimulated HRMCs. Notably, circ-GNB4 could target miR-23c and EGR1 was targeted by miR-23c, as confirmed by dual-luciferase reporter assay and RNA immunoprecipitation. Moreover, EGR1 expression was positively modulated by circ-GNB4 via miR-23c. Collectively, circ-GNB4 might be a novel mechanism of DN-induced HRMCs injury, and there was a circ-GNB4/miR-23c/EGR1 pathway underlying the proliferation, extracellular matrix accumulation, inflammation and oxidative stress. This study suggested circ-GNB4 as a potential target to interfere the development of DN.

Стан системи антиоксидантного захисту у риб Danio rerio за інтоксикації хлорпірифосом

The article presents the results of studies of acute intoxication in Danio Rerio fish by chlorpyrifos at concentrations 0.5 mg/L; 0.75 mg/L and 1 mg/L of aquarium water for 24 h and its influence on the content of TBK-active products, catalase activity (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPO), glutathione reductase (GR) in their tissues. After 24 h in the fish staying in water with chloropyrifos added in concentrations 0.75 mg/L and 1 mg/L we have revealed an increase of the content of TBK-active products in the studied tissues compared to the control values. The linear nature of the growth of the CAT enzymatic activity in the tissues of head and torso due to the effects of chloropyrifos in doses of 0.5 mg/L; 0.75 mg/L and 1 mg/L. was shown. On the background of the catalase activity growth in Danio rerio, in 24 h after the action of the toxicant, there was an inversely proportional dependence between the dose of chlorpyrifos and the activity of GPOs in both investigated parts of the fish body. In the tissues of the head and torso, inhibition of SOD activity for the action of chlorpyrifos has been established. The activity of this enzyme was the lowest at the chlorpyrifos concentration 0.75 mg/L. The obtained results complement the data on the toxic effect of chlorpyrifos on ichthyofauna and on the course of chlorpyrifos-induced oxidative stress in Danio rerio fish. The obtained data can be used in the development of additional biochemical markers of chlorpyrifos and other phosphor organic compounds intoxication and in toxicological and environmental experiments, taking into account the growing intensity of the use of insecticidal preparations in agricultural production and everyday life.

DIP2A is involved in SOD-mediated antioxidative reactions in murine brain

Autism spectrum disorders (ASDs) are highly associated with oxidative stress. We have recently shown that Disconnected-interacting protein homolog 2 A (DIP2A) functions in ASD pathophysiology by regulating cortactin acetylation for spine development and synaptic transmission. However, its role is not fully understood in the context of its abundant expression in mitochondria. In this paper, we found that DIP2A was involved in superoxide dismutase (SOD)-mediated antioxidative reactions. In mice, DIP2A knockout inhibited SOD activity and increased reactive oxygen species (ROS) levels in the cerebral cortex. In vitro gain-of-function experiments further confirmed the positive role of DIP2A in scavenging ROS upon oxidative stress. Moreover, DIP2A knockout caused irregular mitochondrial morphology in the cerebral cortex and impaired mitochondrial metabolism with an over consumption of lipids for energy supply. Taken together, these results revealed unrecognized functions of DIP2A in antioxidative protection, providing another possible explanation for DIP2A-mediated ASD pathophysiology.

EFFECT OF COMPOSITE DRUG ETHYLTHIOSULPHONATE WITH SURFACTANT RAMNOLIPID BIOCOMPLEX ON SH-GROUP CONTENT IN DAPHNIA MAGNA

One of the main obstacles to the use of thiosulfonates in aquaculture is their insolubility in water. This problem is eliminated by using a composition based on ethylthiosulfanilate (ETS) and surface-active rhamnolipid biocomplex (RBC) of bacteria of the genus Pseudomonas, which provides increased bioavailability of ethylthiosulfanilate and activation of biocidal properties of the drug. However, this may increase the toxic effects of the created composition. Given the possibility of using Daphnia magna Straus in the procedure of bioencapsulation as a vector organism for the delivery of a composite drug, a study of the effect of ETS: RBC in nontoxic concentrations (2.5·10-3 mg / ml and 5·10-3 mg / ml (by ETS) on the content of protein and non-protein thiols in Daphnia magna, as well as the activity of enzymes inhibited by a natural analogue of thiosulfonates - allicin. A decrease in the content of protein and non-protein thiols in the tissues of Daphnia magna was found under the action of both studied concentrations of the drug. A significant reduction in non-protein SH groups compared to protein-bound ones may indicate the active and efficient involvement of glutathione ethylthiosulfanilate, which is the predominant non-protein thiol in the cell and provides protection against oxidation of functional groups of proteins and enzymes. The established inhibition of superoxide dismutase activity may be due to S-thioallylation of ethylthiosulfanilate SH-containing amino acids in the structure of the enzyme. No significant changes in glutathione-S-transferase activity under the action of the study drug were observed. Only when using a higher concentration of the composite drug ETS: RBC begins to show a tendency to decrease activity, most likely due to a decrease in the content of reduced glutathione under these conditions. Given the survival rates of Daphnia magna under the conditions of using the composite drug ETS: RBC at concentrations of 2.·10-3 mg / ml and 5·10-3 mg / ml, it is likely that its effect focuses on reactions involving sulfhydryl groups and does not cause significant changes in the detoxification system of the cell.