Intracellular Antioxidant Capacity Research Articles

Efficient water-soluble copper(II) complexes with SOD/CO-like activities and the relationship between their structure and activity

Superoxide dismutase is a significant member of the body’s natural antioxidant defense system. It can effectively eliminate superoxide anion free radicals from the body. Based on the functionalized ligand-isonicotinic acid, three copper complexes were successfully developed. The metal centers of complexes 1–3 were highly similar to the active sites of natural superoxide dismutase. It was found that complexes 1–3 exhibited both excellent superoxide dismutase (SOD)-like and catecholase (CO)-like activities at the same time. A model of oxidative stress, induced by hydrogen peroxide, was constructed based on the cytotoxicity test. In this model, the mimetic complex proved to have effective intracellular antioxidant capacity. Combined with the experimental data and DFT, the results showed that the electron acceptance ability and the coordination configuration of the active center of copper ions had important effects on the activity of SOD.

Rapid Adaptability and Regulation of Physiological Status of Lager Yeast Ensure Its Activity During Dehydration

Yeast products either active dry yeast or liquid yeast are used for beer brewing. Dry yeast offers advantages over liquid yeast, including longer shelf life, easier storage and transportation, convenient usage. While active dry lager yeast now has gained popularity in Chinese breweries, lacking knowledge about yeast physiology status during drying process hinders the customizing dry yeast for specific requirements. This study delved into the dehydration process of two lager yeasts G03 and M14 used in Chinese breweries and one commercial yeast W34/70. W34/70 with greater desiccation resistance exhibiting rapid adaptation during drying process maintained highest intracellular adenosine-5′-triphosphate (ATP). Improved accumulation of antioxidants including glutathione and trehalose, as well as complementary synthesis of chitin in cell wall attributed to the desiccation resistance of yeasts. Higher intracellular energy supply and anti-oxidative capacity of yeast strain facilitated the balanced reactive oxygen species (ROS) accumulation. Together with an effective DNA synthesis and cell replication, more cells would survive after dehydration and subsequent rehydration. Current study provides new evidence for the theory of yeast desiccation resistance and further help with preparation of active dry yeast products.

N6022 attenuates cerebral ischemia/reperfusion injury-induced microglia ferroptosis by promoting Nrf2 nuclear translocation and inhibiting the GSNOR/GSTP1 axis

Stroke poses a significant risk of mortality, particularly among the elderly population. The pathophysiological process of ischemic stroke is complex, and it is crucial to elucidate its molecular mechanisms and explore potential protective drugs. Ferroptosis, a newly recognized form of programmed cell death distinct from necrosis, apoptosis, and autophagy, is closely associated with the pathophysiology of ischemic stroke. N6022, a selective inhibitor of S-nitrosoglutathione reductase (GSNOR), is a “first-in-class” drug for asthma with potential therapeutic applications. However, it remains unclear whether N6022 exerts protective effects in ischemic stroke, and the precise mechanisms of its action are unknown. This study aimed to investigate whether N6022 mitigates cerebral ischemia/reperfusion (I/R) injury by reducing ferroptosis and to elucidate the underlying mechanisms. Accordingly, we established an oxygen-glucose deprivation/reperfusion (OGD/R) cell model and a middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model to mimic cerebral I/R injury. Our data, both in vitro and in vivo, demonstrated that N6022 effectively protected against I/R-induced brain damage and neurological deficits in mice, as well as OGD/R-induced BV2 cell damage. Mechanistically, N6022 promoted Nrf2 nuclear translocation, enhancing intracellular antioxidant capacity of SLC7A11-GPX4 system. Furthermore, N6022 interfered with the interaction of GSNOR with GSTP1, thereby boosting the antioxidant capacity of GSTP1 and attenuating ferroptosis. These findings provide novel insights, showing that N6022 attenuates microglial ferroptosis induced by cerebral I/R injury through the promotion of Nrf2 nuclear translocation and inhibition of the GSNOR/GSTP1 axis.

Astragalus polysaccharides attenuate rat aortic endothelial senescence via regulation of the SIRT-1/p53 signaling pathway

BackgroundAstragalus polysaccharides (APS) have been verified to have antioxidative and antiaging activities in the mouse liver and brain. However, the effect of APS on aortic endothelial senescence in old rats and its underlying mechanism are currently unclear. Here, we aimed to elucidate the effects of APS on rat aortic endothelial oxidative stress and senescence in vitro and in vivo and investigate the potential molecular targets.MethodsTwenty-month-old natural aging male rats were treated with APS (200 mg/kg, 400 mg/kg, 800 mg/kg daily) for 3 months. Serum parameters were tested using corresponding assay kits. Aortic morphology was observed by staining with hematoxylin and eosin (H&E) and Verhoeff Van Gieson (VVG). Aging-related protein levels were evaluated using immunofluorescence and western blot analysis. Primary rat aortic endothelial cells (RAECs) were isolated by tissue explant method. RAEC mitochondrial function was evaluated by the mitochondrial membrane potential (MMP) measured with the fluorescent lipophilic cationic dye JC‑1. Intracellular total antioxidant capacity (T-AOC) was detected by a commercial kit. Cellular senescence was assessed using senescence-associated-β-galactosidase (SA-β-Gal) staining.ResultsTreatment of APS for three months was found to lessen aortic wall thickness, renovate vascular elastic tissue, improve vascular endothelial function, and reduce oxidative stress levels in 20-month-old rats. Primary mechanism analysis showed that APS treatment enhanced Sirtuin 1 (SIRT-1) protein expression and decreased the levels of the aging marker proteins p53, p21 and p16 in rat aortic tissue. Furthermore, APS abated hydrogen peroxide (H2O2)-induced cell senescence and restored H2O2-induced impairment of the MMP and T-AOC in RAECs. Similarly, APS increased SIRT-1 and decreased p53, p21 and p16 protein levels in senescent RAECs isolated from old rats. Knockdown of SIRT-1 diminished the protective effect of APS against H2O2-induced RAEC senescence and T-AOC loss, increased the levels of the downstream proteins p53 and p21, and abolished the inhibitory effect of APS on the expression of these proteins in RAECs.ConclusionAPS may reduce rat aortic endothelial oxidative stress and senescence via the SIRT-1/p53 signaling pathway.

Whey-pectin microcapsules improve the stability of grape marc phenolics during digestion.

Grape marc (GM) is an agri-food residue from the wine industry valuable for its high content of phenolic compounds. This study aimed to develop an encapsulation system for GM extract (GME) using food-grade biopolymers resistant to gastric conditions for its potential use as a nutraceutical. For this purpose, a hydroalcoholic GME was prepared with a total phenolics content of 219.62±11.50mg gallic acid equivalents (GAE)/g dry extract and 1389.71±97.33µmol Trolox equivalents/g dry extract antioxidant capacity, assessed through ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay. Moreover, the extract effectively neutralized reactive oxygen species in Caco-2 cells, demonstrating an intracellular antioxidant capacity comparable to Trolox. The GME was encapsulated using whey protein isolate and pectin through nano spray drying (73% yield), resulting in spherical microparticles with an average size of 1±0.5µm and a polydispersity of 0.717. The encapsulation system protected the microcapsules from simulated gastrointestinal digestion (GID), where at the end of the intestinal phase, 82% of the initial phenolics were bioaccessible compared to 54% in the free GME. Besides, the encapsulated GME displayed a higher antioxidant activity by the ferric reducing antioxidant power assay than the free extract after GID. These results show the potential of this encapsulation system for applying GME as a nutraceutical with a high antioxidant capacity and protective effect against cellular oxidation.

Natural product piperlongumine inhibits proliferation of oral squamous carcinoma cells by inducing ferroptosis and inhibiting intracellular antioxidant capacity.

As a new form of cell death, ferroptosis has been shown to have inhibitory effects on a variety of tumor cells except oral squamous cell carcinoma (OSCC). There were few investigations on the effects and molecular mechanisms of piperlongumine (PL, a ferroptosis inducer) and CB-839 (a GLS1 inhibitor which promotes ferroptosis) on OSCC cells. This article assesses the anticancer effect and mechanism of PL as well as combined with CB-839. OSCC cells were treated with specified concentration of PL alone or with ferroptosis inhibitor Ferrostatin-1 (Fer-1) and antioxidant N-Acetylcysteine (NAC) to assess their effects on biological characteristics such as cell proliferation, cell death and intracellular ferroptosis related pathways. Also, cells were treated with PL combined with CB-839 to evaluate the synergistic effect of CB-839 on PL's anticancer effects. The results showed that the proliferation rate of PL-treated OSCC cells were decreased in a dose- and time-dependent manner. PL can induce OSCC cells apoptosis. Lipid peroxidation (LPO) and intracellular reactive oxygen species (ROS) were accumulated after PL treatment. We found some protein changes significantly such as the expression of DMT1 increased, and the expression of FTH1, SLC7A11 and GPX4 decreased. In addition, the anti-proliferation effect of PL can be reversed by Fer-1 and NAC and the level of LPO and ROS was decreased accordingly. Importantly, we found that PL and CB-839 in combination could decrease the cell viability and the LPO level synergistically, accompanied by a large consumption of glutathione (GSH). These evidences prove that PL can induce ferroptosis of OSCC cells, which can be enhanced by CB-839. Our study suggested that the nature product PL can induce the ferroptotic death of OSCC cells, which is further enhanced when combined with CB-839. The synergistic anticancer effect of these two may prove new strategy for OSCC treatment.

Effect of pulsed light on curcumin chemical stability and antioxidant capacity.

Curcumin is the major bioactive component in turmeric with potent antioxidant activity. Little is known about how pulsed light (PL) technology (an emerging non-thermal food processing technology relying on high intensity short duration flashes of light) can affect the chemical stability and antioxidant capacity of curcumin. This study found that PL treatment of fluence levels from 0 to 12.75 J/cm2 produced a fluence-dependent reduction in curcumin content. These results paralleled the production of a tentative curcumin dimer, identified as a potential photochemical transformation product. PL-treated curcumin at relatively higher fluence levels decreased chemical-based ORAC and ABTS antioxidant capacity, relative to control (P < 0.05). This contrasted the effect observed to increase coincidently both intracellular antioxidant capacity (e.g., DCFH-DA (P < 0.05)) and GSH/GSSG ratio (P < 0.05), respectively, in cultured differentiated Caco-2 cells. In conclusion, the application of PL on curcumin results in photochemical transformation reactions, such as dimerization, which in turn, can enhance biological antioxidant capacity in differentiated Caco-2 cells.

Influence of sprouting on the bioaccessibility and bioactivity of benzoxazinoids, phenolic acids, and flavonoids of soft and hard wheat cultivars

Grain germination increases the contents of benzoxazinoids and the antioxidant capacity of wheat and differentially affects the phytochemical composition of hard and soft wheat cultivars. It was investigated whether wheat cultivars (sprouted or not) with distinct grain texture characteristics (BRS Guaraim, soft vs. BRS Marcante, hard texture) would have different behavior in relation to changes in phytochemical compounds, bioaccessibility and antioxidant capacity during simulated gastrointestinal digestion of a tabbouleh preparation. Sprouting increased the nominal amount of phytochemicals in tabbouleh resulting in increased release of phenolic acids (up to 7.5-fold) and benzoxazinoids (up to 12.5-fold) during all digestion phases besides higher bioaccessibility (up to 2.8-fold). Sprouting caused greater increase in the bioaccessibility of phenolic acids for the soft wheat cultivar (4.5-fold) than for the hard cultivar (1.9-fold) and it increased the colon available index of phenolic acids only for the soft cultivar (1.8-fold). Flavonoids, mainly represented by apigenin glycosides, were marginally increased after sprouting but underwent relative increase along digestion being the major phytochemicals found in the bioaccessible fraction obtained after intestinal digestion (73–94% of total phytochemicals). The increase in apigenin glycosides was associated to the increase of in vitro and intracellular antioxidant capacity of tabbouleh along digestion. Sprouting increased the peroxyl radical removal capacity of tabbouleh in the gastric phase and in the non-bioaccessible fraction regardless of the cultivar. The highest hydroxyl radical removal capacities were found in non-sprouted cultivars, especially in the soft texture cultivar in the undigested and bioaccessible fractions. The bioaccessible fraction obtained after wheat digestion was more efficient to scavenge intracellular ROS than undigested samples, the highest scavenging potency being observed for the hard texture cultivar with no effect of sprouting. These findings confirm the hypothesis that the phytochemicals of hard and soft wheat cultivars (sprouted or not) have different behavior during digestion in terms of biotransformation, bioaccessibility and ability to remove reactive species and indicate that tabbouleh produced from sprouted wheat results in increased release of bioactive phytochemicals during digestion.

Looking beyond Arthrospira: Comparison of antioxidant and anti-inflammatory properties of ten cyanobacteria strains

Cyanobacteria are a widely distributed group of photosynthetic prokaryotes with potential to produce biologically active compounds. Arthrospira is the most widespread grown cyanobacteria for commercial purposes; however, many other species could be used as sources of a wide variety of bio-products. The pigment phycocyanin explains part of the therapeutic properties attributed to Arthrospira (e.g. antioxidant and anti-inflammatory). Many chronic diseases include, on its etiology, oxidative stress and abnormalities in the inflammatory response; thus, there is great interest in finding new sources of natural antioxidants and anti-inflammatory compounds. The aim of this work was to evaluate growth attributes, chemical composition and antioxidant and anti-inflammatory properties of ten cyanobacteria isolates. After 15 days of culture, biomass production ranged between 0.06 g/L (Anabaena) and 0.27 g/L (Phormidium) in 20 L volume cultures. Protein content of dry biomass ranged between 28.86 % (Nodularia) and 56.39 % (Arthrospira) while phycocyanin content ranged between 0.86 % (Nodularia) and 6.66 % (Spirulina). Aqueous extracts were evaluated regarding total phenolic content (TPC), total flavonoids content (TFC) and antioxidant activities (DPPH, ABTS and FRAP). Spirulina and Desertifilum exhibited the highest phenols and flavonoids contents; as well as the highest ABTS and FRAP antioxidant activities. Five strains were selected for measuring anti-inflammatory activity by the human erythrocyte membrane stabilization assay. All the strains evaluated exhibited a remarkable anti-inflammatory capacity, being the highest value for Desertifilum. Strains of Spirulina and Desertifilum were also tested for their ability to scavenge reactive oxygen species (ROS) on Hek293 cells; both strains showed significant intracellular antioxidant capacity and were not cytotoxic. This research demonstrated that other cyanobacterial strains, besides the popular Arthrospira, might be new sources of antioxidant and anti-inflammatory compounds for biotechnological applications in functional food, nutraceutical or pharmaceutical industries.

The Bioactive Properties of Carotenoids from Lipophilic Sea buckthorn Extract (Hippophae rhamnoides L.) in Breast Cancer Cell Lines.

In women, breast cancer is the most commonly diagnosed cancer (11.7% of total cases) and the leading cause of cancer death (6.9%) worldwide. Bioactive dietary components such as Sea buckthorn berries are known for their high carotenoid content, which has been shown to possess anti-cancer properties. Considering the limited number of studies investigating the bioactive properties of carotenoids in breast cancer, the aim of this study was to investigate the antiproliferative, antioxidant, and proapoptotic properties of saponified lipophilic Sea buckthorn berries extract (LSBE) in two breast cancer cell lines with different phenotypes: T47D (ER+, PR+, HER2-) and BT-549 (ER-, PR-, HER2-). The antiproliferative effects of LSBE were evaluated by an Alamar Blue assay, the extracellular antioxidant capacity was evaluated through DPPH, ABTS, and FRAP assays, the intracellular antioxidant capacity was evaluated through a DCFDA assay, and the apoptosis rate was assessed by flow cytometry. LSBE inhibited the proliferation of breast cancer cells in a concentration-dependent manner, with a mean IC50 of 16 µM. LSBE has proven to be a good antioxidant both at the intracellular level, due to its ability to significantly decrease the ROS levels in both cell lines (p = 0.0279 for T47D, and p = 0.0188 for BT-549), and at the extracellular level, where the ABTS and DPPH inhibition vried between 3.38-56.8%, respectively 5.68-68.65%, and 35.6 mg/L equivalent ascorbic acid/g LSBE were recorded. Based on the results from the antioxidant assays, LSBE was found to have good antioxidant activity due to its rich carotenoid content. The flow cytometry results revealed that LSBE treatment induced significant alterations in late-stage apoptotic cells represented by 80.29% of T47D cells (p = 0.0119), and 40.6% of BT-549 cells (p = 0.0137). Considering the antiproliferative, antioxidant, and proapoptotic properties of the carotenoids from LSBE on breast cancer cells, further studies should investigate whether these bioactive dietary compounds could be used as nutraceuticals in breast cancer therapy.

The Role of G3BP1 Gene Mediates P38 MAPK/JNK Pathway in Testicular Spermatogenic Dysfunction Caused by Cyfluthrin.

In recent years, male infertility has received global attention and seriously affected the quality of human fertility, and pyrethroids (type II pyrethroids), as recognized environmental endocrine disruptors, may threaten male reproductive health. Therefore, in this study, we established an in vivo model for the development of testicular and germ cell toxicity induced by cyfluthrin and explored the role and mechanism of the G3BP1 gene-mediated P38 MAPK/JNK pathway in testicular and germ cell damage caused by cyfluthrin to find early and sensitive indicators and new therapeutic targets for the development of testicular damage. Firstly, 40 male Wistar rats (about 260 g) were divided into a control group (corn oil), low dose group (6.25 mg/kg), middle dose group (12.5 mg/kg) and high dose group (25 mg/kg). The rats were anesthetized and executed after 28 days of poisoning on alternate days. Then, HE staining, transmission electron microscopy, ELISA, q-PCR, Western blot, immunohistochemistry, double-immunofluorescence and TUNEL were used to observe the pathology, androgen levels, oxidative damage and altered expression of the key factors of the G3BP1 and MAPK pathways in rat testes. The results showed that, compared with the control group, the testicular tissue and spermatocytes were superficially damaged with an increasing dose of cyfluthrin; furthermore, it could interfere with the normal secretion of the hypothalamic-pituitary-gonadal axis (serum GnRH, FSH, T and LH levels) and cause hypergonadal dysfunction. A dose-dependent increase in MDA and a dose-dependent decrease in T-AOC indicated that the oxidative-antioxidative homeostatic balance was disrupted. The Western blot and qPCR analysis revealed that G3BP1, p-JNK1/2/3, P38 MAPK, p-ERK, COX1 and COX4 proteins and mRNA expression were decreased, and p-JNK1/2/3, p-P38MAPK, caspase 3/8/9 proteins and mRNA expression were significantly increased. The double-immunofluorescence and immunohistochemistry results showed that the protein expression of G3BP1 decreased with an increasing dose of staining, while the expression of JNK1/2/3 and P38 MAPK were increased significantly. The positive expressions of G3BP1 were mainly located in the testicular germinal epithelium and germ cell layer, and the positive expressions of JNK1/2/3 were mainly located in the testicular germinal epithelium and sperm cells, while the positive expressions of P38 MAPK were located in all levels of the germ cells and spermatozoa. Our results demonstrated that exposure to cyfluthrin caused testicular and spermatocyte damage in rats, which could cause pathomorphology, altered androgen levels and a decreased antioxidant capacity. When the intracellular antioxidant capacity was impaired, G3BP1 expression and activity were inhibited, causing activation of the P38 MAPK/JNK pathway and activation of the intracellular apoptotic pathway, which, in turn, led to germ cell apoptosis.

In vitro antioxidant and lipid-lowering properties of free and bound phenolic compounds from buckwheat hulls

Polyphenols can perform preventive and alleviating functions in many metabolic disorders. Buckwheat hulls, usually discarded as waste, are known to have plenty of phenolic compounds. The present study aimed to explore the composition and in vitro lipid-lowering and antioxidant activities of the free and bound phenolic compounds (FPBH and BPBH) in buckwheat hulls. The contents of phenolic compounds in FPBH and BPBH were 150.25 ± 6.25 mg g−1 and 44.25 mg g−1 respectively, indicating that free phenols were the main form of phenols in buckwheat hulls. Rutin and syringic acid was the most abundant phenolic compounds in FPBH and BPBH respectively. The results of DPPH and ABTS free radical scavenging ability showed that the antioxidant activity of FPBH was significantly higher than that of BPBH. In vitro assessments showed that both FPBH and BPBH could significantly alleviate lipid accumulation and oxidative stress in oleic acid (OA)-treated HepG2 cells with a dose-dependent manner by decreasing the levels of intracellular ROS, mitochondrial membrane potential (MMP), triglyceride (TG), total cholesterol (TC) and low density lipoprotein (LDL-C) as well as significantly improving intracellular total antioxidant capacity (TAC), the levels of high density lipoprotein (HDL-C) and activities of glutathione peroxidase.(GSH-Px), superoxide dismutase (SOD) and catalase (CAT). Meanwhile, FPBH was more effective than BPBH at the same concentration. It was concluded that phenolic compounds in buckwheat hulls might have great potential to develop functional foods in improving oxidative damage and lipid metabolism disorders.

Engineering a synergistic antioxidant inhibition nanoplatform to enhance oxidative damage in tumor treatment.

The antioxidant system of tumor cells severely impairs reactive oxygen species (ROS)-mediated tumor therapy. Despite extensive attempts to attenuate the antioxidant capacity by eliminating ROS scavengers such as glutathione (GSH), nicotinamide adenine dinucleotide phosphate (NADPH) over-expressed in the tumor microenvironment can regenerate GSH from glutathione disulfide (GSSG), hence weakening ROS-induced oxidative damage. Therefore, engineering a nanoplatform capable of depleting both NADPH and GSH is extremely significant for improving ROS-mediated tumor treatment. Herein, a synergetic antioxidant inhibition strategy is proposed to attenuate intracellular antioxidant capacity for hypoxic tumor therapy. In this context, both porous Prussian blue nanoparticles (PPB NPs) and cisplatin prodrug [cis-Pt (IV)] in the nanoplatform can oxidize GSH to directly reduce GSH levels, while PPB NPs also enable NADPH depletion by peroxidase-mimicking to impair GSH regeneration. Furthermore, PPB NPs with catalase-mimicking activity catalyze H2O2 decomposition to alleviate tumor hypoxia, thus reducing the generation of GSH and boosting singlet oxygen (1O2) production by Chlorin e6 (Ce6) for enhancing oxidative damage. Experimental results prove that the nanoplatform, denoted as PPB-Ce6-Pt, can induce remarkable tumor cells apoptosis and ferroptosis. Importantly, a simple loading method and the use of Food Drug Administration (FDA)-approved materials make PPB-Ce6-Pt have great potential for practical applications. STATEMENT OF SIGNIFICANCE: The antioxidant system in tumor cells disables ROS-mediated tumor therapy. Besides, extensive attempts aim at depleting GSH without considering their regeneration. Therefore, we developed a synergetic strategy to attenuate intracellular antioxidant capacity for hypoxic tumor therapy. PPB-Ce6-Pt nanoplatform could not only directly reduce GSH levels but also deplete NADPH by peroxidase-mimicking to impair GSH regeneration. In addition, PPB-Ce6-Pt nanoplatform could catalyze H2O2 decomposition to alleviate tumor hypoxia, thus reducing the generation of GSH and boosting 1O2 production by Chlorin e6 (Ce6) for increasing oxidative damage. Then, intracellular ROS boost and redox dyshomeostasis induced remarkable tumor cells apoptosis and ferroptosis. Importantly, a simple loading method and the use of biosafety materials made the nanoplatform have great potential for practical applications.

Multi-scale analysis of nickel ion tolerance mechanism for thermophilic Sulfobacillus thermosulfidooxidans in bioleaching.

Bioleaching is intensively investigated for recovering valuable metals such as Li, Co, Ni and Cu. Nickel ion stress threatens the health of microorganisms when Ni2+ starts to accumulate in the leachate during the bioleaching of materials that are rich in Ni, such as spent lithium-ion batteries. The possible mechanisms underlying the response of S. thermosulfidooxidans to nickel ion stress were analyzed using a multi-scale approach. Under the condition of nickel ion stress, high concentrations of nickel ions were immobilized by extracellular polymeric substances, while concentrations of nickel ions inside the cells remained low. The intracellular adenosine triphosphate (ATP) concentration and H+-ATPase activity increased to maintain normal cell growth and metabolic activities. Scavenging abilities of S. thermosulfidooxidans for hydrogen peroxide and superoxide anion were enhanced to reduce oxidative damage induced by nickel ion stress. There were 734 differentially expressed genes identified by RNA-seq under nickel ion stress. Most of them were involved in oxidative phosphorylation, glutathione metabolism and genetic information processing, responsible for intracellular energy utilization, intracellular antioxidant capacity and DNA damage repair, respectively. The results of this study are of major significance for in-depth understanding of the mechanisms of acidophilic microorganisms' resistance to metal ions.

A dual-responsive ratiometric indicator designed for in vivo monitoring of oxidative stress and antioxidant capacity.

The imbalance between oxidative stress and antioxidant capacity is strongly associated with the development of numerous degenerative diseases, including cardiovascular diseases, diabetes, neurodegenerative diseases, and cancer. Therefore, monitoring oxidative stress and antioxidant capacity in vivo is crucial for maintaining cellular homeostasis and the stability of the organism's internal environment. Here, we present the findings of our study on DQ1, a dual-responsive indicator designed specifically for imaging H2O2 and NAD(P)H, which are critical indicators of oxidative stress and antioxidant capacity. DQ1 facilitated the colorimetric and fluorescence detection of H2O2 and NAD(P)H in two well-separated channels, exhibiting a detection limit of 1.0 μM for H2O2 and 0.21 nM for NAD(P)H, respectively. Experiments conducted on living cells and zebrafish demonstrated that DQ1 could effectively detect changes in H2O2 and NAD(P)H levels when exposed to exogenous hypoxic conditions and chemical stimuli. Furthermore, the effectiveness of the as-fabricated indicator was investigated in two distinct mouse models: evaluating H2O2 and NAD(P)H levels in myocardial cell dysfunction during acute myocardial infarction and liver tissue damage under trichloroethylene stress conditions. In vivo experiments demonstrated that the levels of the two cardiac biomarkers increase progressively with the development of myocardial infarction, eventually reaching a steady state after 7 days when the damaged cells in the infarcted region become depleted. Moreover, during 14 continuous days of exposure to trichloroethylene, the two biomarkers in liver tissue exhibited a sustained increase, indicating a significant enhancement in intracellular oxidative stress and antioxidant capacity attributed to the mouse liver's robust metabolic capacity. The aforementioned studies underscore the efficacy of DQ1 as a valuable tool for scrutinizing redox states at both the single-cell and biological tissue levels. It presents significant potential for investigating the dynamic alternations in oxidative stress and antioxidant capacity within disease models as the disease progresses, thereby facilitating a more profound comprehension of these processes across various disease models.

Effect of micro-nanobubbles with different gas sources on the growth and metabolism of chemoautotrophic microorganisms

Micro-nanobubbles (MNBs) have excellent stability and a long retention time in water; their properties vary greatly with different gas sources. In this study, the effects of MNBs with different gas sources, including CO2, N2, O2, and air, on growth and metabolism of the chemoautotrophic strain Sinomicrobium oceani WH-15, which fixes CO2 via the Calvin-Benson-Bassham pathway, were evaluated. MNBs at low concentrations (5–20 %) could promote microbial growth with a maximum increase of 55 % (air MNBs) whereas MNBs at high concentrations (20–50 %) inhibited microbial growth with a maximum decrease of − 79 % (CO2 MNBs). The influence of MNBs was achieved through indirect and direct roles. MNBs with different gas sources had different effects on physical and chemical factors (pH, dissolved oxygen, oxidation-reduction potential, and conductivity) of microbial culture medium. MNBs at high concentration led to an increased intracellular antioxidant capacity of microorganisms and a decrease in the intracellular reduced glutathione, which may be due to exogenous reactive oxygen species produced by MNBs. MNBs could bring reducing power to microorganisms and reduce their metabolic output of extracellular polysaccharides. Our results demonstrated that MNBs with different gas sources improved microbial CO2 fixation through different mechanisms.

Enhancing Bioactive Components of Euryale ferox with Lactobacillus curvatus to Reduce H2O2-Induced Oxidative Stress in Human Skin Fibroblasts.

This study investigated the effects of Lactobacillus curvatus fermentation on the oxidative stress attenuating effects of Euryale ferox on H2O2-induced human skin fibroblasts (HSF). The results showed that Lactobacillus curvatus fermentation (i) increases the content of the various bioactive components of Euryale ferox and is found to have smaller molecular weights of polysaccharides and polypeptides; (ii) increases the overall intracellular and extracellular antioxidant capacity of H2O2-induced HSF while reducing reactive oxygen species (ROS) levels. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) all showed simultaneous increases in activity. Aside from that, the Nrf2 and MAPK signaling pathways are activated to regulate downstream-associated proteins such as the Bax/Bcl-2 protein ratio, matrix metalloproteinase 1 (MMP-1) activity, and human type I collagen (COL-1). These results suggested that the fermentation of Euryale ferox with Lactobacillus curvatus enhances its antioxidant capacity and attenuates apoptosis and senescence caused by oxidative stress.

Maternal Dietary Supplementation with γ-Aminobutyric Acid Alleviated Oxidative Stress in Gestating Sows and Their Offspring by Regulating GABRP.

Simple SummarySows usually suffer oxidative stress during gestation, and this limits the growth of fetuses via placenta. Gamma-aminobutyric acid (GABA) is a functional nonessential amino acid engaged in regulating the physiological status of animals. However, the effects of GABA on the oxidative homeostasis of sows and their offspring remain unclear. This study aims to investigate the effects of GABA on the oxidative stress of gestating sows, newborn piglets, as well as the H2O2-treated porcine trophectoderm cell line-2 (pTr-2 cells) and to subsequently provide guidance on the application of GABA in sows’ nutrition during late gestation. We found that GABA alleviated placental oxidative stress in gestating sows by increasing antioxidant-associated enzyme activity and by upregulating the gene expressions of gamma-aminobutyric acid receptor (GABRP) and nuclear factor-erythroid 2-related factor-2 (Nrf2) in pTr-2 cells, as well as enhanced the antioxidant capacity of sows and fetal piglets. This study provides the possibility for future application of GABA in maternal–fetal nutrition.Sows usually suffer oxidative stress during gestation, and this limits the growth of fetuses via placenta. Gamma-aminobutyric acid (GABA) is a functional nonessential amino acid engaged in regulating the physiological status of animals. However, the effects of GABA on the oxidative homeostasis of sows and their offspring remain unclear. Eighteen late gestating sows (85 d) were divided into the CON and GABA groups and fed the basal diet and the GABA diet (200 mg/kg GABA), respectively, until farrowing. At parturition, the sows’ litter characteristics, the plasma antioxidant parameters of sows, and their offspring were evaluated. The results showed that GABA supplementation had no marked effect on the reproductive performance of sows (p > 0.10) but had a trend of reducing the amount of intrauterine growth restriction (IUGR) in piglets (0.05 < p < 0.10). At the same time, the addition of GABA elevated the plasma superoxide dismutase (SOD) level of sows and enhanced the glutathione peroxidase (GSH-Px) activity of newborn piglets (p < 0.05). Based on the H2O2-induced oxidative stress in pTr-2 cells, GABA elevated intracellular GSH-Px, SOD, catalase (CAT), and total antioxidant capacity (T-AOC, p < 0.01) and upregulated the gene expressions of CAT, gamma-aminobutyric acid receptor (GABRP), and nuclear factor-erythroid 2-related factor-2 (Nrf2) in H2O2-treated pTr-2 cells (p < 0.05). Taken together, GABA improved the antioxidant capacity of sows and alleviated the placental oxidative stress by upregulating the GABRP and Nrf2 genes, which have the potential to promote oxidative homeostasis in newborn piglets.

Effect of radio frequency, ultrasound, microwave-assisted papain, and alcalase hydrolysis on the structure, antioxidant activity, and peptidomic profile of Rosa roxburghii Tratt. seed protein.

Traditional enzymatic hydrolysis methods have defects such as low efficiency and poor bioactivity in the production of active peptides. In this study, radio frequency (RF) technology was innovatively used to assist the hydrolysis of Rosa roxburghii Tratt. seed protein (RTSP) by papain and alcalase. RF-assisted hydrolysis was compared with ultrasound-(US) and microwave (MW)-assisted techniques in terms of the degree of hydrolysis, structure, antioxidant properties, and changes in the peptidome of the hydrolysates to clarify the mechanism of functional change of physically-assisted hydrolysate. All three methods improved hydrolysis efficiency. The degree of hydrolysis (DH) of papain group increased from 6.38% to 7.97%, 9.97% and 8.37% after US-, MW- and RF-assisted hydrolysis, respectively, while the DH of alcalase-treated group increased from 21.13% to 25.66%, 26.03%, and 23.01%, respectively. The in vitro antioxidant capacity and intracellular antioxidant capacity of RTSP and its hydrolysates were measured and evaluated by fuzzy statistical evaluation, and MW-assisted alcalase hydrolysis had the highest in vitro and intracellular antioxidant activity scores of 0.713 and 0.820, respectively. Fourier transform infrared and amino acid composition analysis explained the enhanced antioxidant properties of the hydrolysates. Further peptide profiling showed the physical assistance led to an increase in the species and contents of small molecule antioxidant peptides compared to enzyme treatment alone. Pearson's linear correlation analysis showed that AY, LY, IY, PHW, SVL, LHL, YYV, VYY, and NHAV were significantly correlated with the antioxidant properties of hydrolysates. Our data suggested that physical assistance such as US, MW, and RF were effective to improve the efficiency of enzymatic hydrolysis and produce novel antioxidant peptides. PRACTICAL APPLICATION: In this study, it was found that electromagnetic wave-assisted enzymatic hydrolysis could improve the efficiency of hydrolysis and enhance the antioxidant activity of hydrolysates compared to unassisted means. Compared with MW treatment, RF has the comparable hydrolysis effect, but has the advantages of high penetration ability, good uniformity, and low energy consumption and has greater potential for the production of bioactive peptides.

Acacetin antagonized lipotoxicity in pancreatic β-cells via ameliorating oxidative stress and endoplasmic reticulum stress.

During the pathogenesis and progression of diabetes, lipotoxicity is a major threat to the function and survival of pancreatic β-cells. To battle against the lipotoxicity induced cellular damages, the present study investigated the beneficial effects of acacetin, a natural antioxidant, on free fatty acid (FFA) stressed RINm5F cells and the potential mechanism involved. RINm5F cells with or without 1h pretreatment of acacetin were treated with 0.35mM sodium palmitate for 24h. Cell viability, intracellular reactive oxygen species (ROS) level, antioxidant capacity, cellular apoptosis, and endoplasmic reticulum (ER) stress biomarker expression were investigated. Our experiments demonstrated that acacetin treatment significantly scavenged the intracellular ROS, upregulated the endogenous antioxidant enzymes, and diminished the sub-G1 DNA fraction in the cells exposed to FFA, suggesting its efficacy against oxidative stress. Meanwhile, acacetin treatment significantly mitigated the overload of intracellular Ca2+ and reduced the pro-apoptotic protein expression in the FFA stimulated cells, and thereby attenuated the ER stress-mediated cell apoptosis. Furthermore, siRNA interference results confirmed that the suppressing of C/EBP-homologous protein (CHOP) was critical to improve FFA-induced reduction in cell viability and ameliorated the ER stress caused by FFA stimulation. Acacetin may antagonize lipotoxicity in pancreatic cells by attenuating the oxidative stress and ER stress.