Antioxidant Gene Expression Research Articles

Molecular characterization of a novel thioredoxin-related transmembrane protein gene AcTMX3 that plays important roles in antioxidant defence in Arma chinensis diapause.

Protein disulphide isomerase (PDI) possesses disulphide isomerase, oxidoreductase and molecular chaperone activities, and is involved in regulating various physiological processes. However, there are few studies on the function in insect diapause. In this study, we cloned one novel member PDI family (TMX3, thioredoxin-related transmembrane protein 3) in Arma chinensis. The AcTMX3 encodes 426 amino acids that contains a predicted N-terminal signal sequence, a thioredoxin-like domain with the CXXC active site and a potential transmembrane region, which are typical sequence features of TMX3. RT-qPCR results showed that AcTMX3 was mainly expressed in the head under non-diapause conditions, while AcTMX3 was highly expressed in the fat body (central metabolic organ) under diapause conditions. Moreover, temporal expression profile showed that compared with non-diapause conditions, diapause conditions significantly induced AcTMX3 expression, and the expression of AcTMX3 was enhanced at 15°C. Silencing AcTMX3 in A. chinensis significantly inhibited the expression of antioxidant genes (AcTrx2 and AcTrx-like), increased the content of H2O2 and ascorbate and reduced the survival rate of A. chinensis under diapause conditions. Our results suggested that AcTMX3 played an important role in the resistance of A. chinensis to oxidative stress under diapause conditions.

Investigating antioxidant effects of hamamelitannin-conjugated zinc oxide nanoparticles on oxidative stress-Induced neurotoxicity in zebrafish larvae model.

An excessive amount of reactive oxygen species triggers oxidative stress, leading to an imbalance in cellular homeostasis. Antioxidant therapy is an effective tool for lowering the oxidative stress and associated ailments. Recently, green nano-based drug formulations have demonstrated promising antioxidant activity and neutralizing oxidative stress. In this study, a tannin molecule Hamamelitannin (HAM), was utilized to synthesize zinc oxide nanoparticles HAM-ZnO NPs, to mitigate oxidative stress and associated ailments . The HAM-ZnO NPs were synthesized and characterized by XRD, SEM, and FTIR. The antioxidant potentials of HAM-ZnO NPs were analyzed by in vitro antioxidant assays. Zebrafish embryos and larvae were used as in-vivo models to assess the toxicity and antioxidant protective mechanism. Hydrogen peroxide (1mM) was employed to induce oxidative stress and treated with HAM-ZnO NPs to study the cognitive impairment and antioxidant enzyme levels. Levels of reactive oxygen species and cell death due to oxidative stress induction were studied by 2',7'-dichlorodihydrofluorescein diacetate and Acridine orange staining methods. Additionally, expression of Antioxidant genes such as SOD, CAT, GPx, and GSR were studied. . HAM-ZnO NPs exhibited a spherical morphology and size ranges between 48 and 53nm. In vitro antioxidant studies revealed the antioxidant properties of HAM-ZnO NPs. Furthermore, in vivo studies indicated that HAM-ZnO NPs don't possess any cytotoxic effects in zebrafish larvae at concentrations between (5-25µg/ml), The study also observed that HAM-ZnO NPs significantly reduced Hydrogen Peroxide-induced stress and increased antioxidant activity in zebrafish larvae. Also, the antioxidant gene expression was upregulated in the HAM-ZnO NPs zebrafish larvae. Findings in this study showed that HAM-ZnO NPs might be a potential intervention for diseases linked to oxidative stress.

Benzo (A) pyrene exposure alters alveolar epithelial and macrophage cells diversity and induces antioxidant responses in lungs

This study was designed to investigate the toxic effects of benzo (a) pyrene (BaP) in the lungs. Mice were repeatedly treated orally with BaP (50 mg/kg body weight, twice a week for four weeks) to induce a tumour. After 4 months of BaP administration, tumours were visible beneath the skin. The histopathological section of the lungs shows congestion of pulmonary blood vessels, alveolar hyperplasia, and concurrent epithelial hyperplasia with infiltrates of inflammatory cells also seen. Thereafter, a single-cell suspension of lung tissues was stained with fluorescently conjugated antibodies for the demarcation of alveolar epithelial (anti-mouse CD74 and podoplanin) and macrophage (F4/80 and CD11b) cells and measured by flow cytometry. The expression of antioxidant genes was assessed by qRT–PCR. The number of alveolar epithelial cells 1 (AEC1) increased, but the number of alveolar epithelial cells 2 (AEC2) and transitional alveolar epithelial cells (TAEC) was significantly decreased in tumour-bearing mice. The proportion of CD11b+ alveolar macrophages (AM) and interstitial macrophages (IM) was increased, but the proportion of F4/80+ AM cells was reduced. The BaP administration significantly increased the ROS production in alveolar cells. The relative expression levels of antioxidant genes (SOD1, catalase, GPX1, and HIF-1α) were increased, but NRF2 expression was decreased in BaP-treated alveolar cells. The expression of anti-inflammatory (NF-κB) was also significantly increased. In conclusion, BaP exposure induced an inflammatory response, altered alveolar epithelial cell and macrophage diversity, and increased antioxidant responses in the lungs.

Effects of dietary rosemary (Rosmarinus officinalis) leaf powder and Bacillus subtilis on the growth performance, digestive enzymes, antioxidant and immune-related gene expression, and resistance to Aeromonas hydriphila in common carp, Cyprinus carpio

Abstract Due to increasing demand about the application of natural feed additives in aquaculture, the efficacy of separate and the combination of rosemary (Rosmarinus officinalis) and B. subtilis on the growth performance, blood indices, innate immunity, antioxidant defense, digestive enzymes activity, the expression levels of genes with immunity modulation, as well as the act of resisting against Aeromonas hydriphila, in common carp, C. carpio is reported. Fish (44.63 ± 0.04 g) were divided into four treatments including the T0 (control treatment) fed with basal diet, T1 fed with rosemary at the rate of 3 %, T2 fed with 1.5×107 CFU g-1 B. subtilis, and T3 fed with a combination of rosemary at the rate of 3 % and 1.5×107 CFU g-1 B. subtilis. After 56 days enriched feed with B. subtilis and combination of rosemary and B. subtilis caused significant improvement in weight gain (WG %), feed conversion ratio (FCR), hematological and biochemical indices, the innate immune responses including in total Ig level, lysozyme and ACH50 activities as well as the chymotrypsin, pepsin, protease, and amylase activities correlate to the control group (P<0.05). Additionally, the serum CAT and GPX activity were significantly enhanced in the T3 group (P<0.05). The mRNA expression of CAT and LYS genes significantly increased in T2 and T3 contrary to T0 (p<0.05). Fish challenged with A. hydrophila had variable survival and the best survival was in the T3. In summary, combination the rosemary and B. subtilis enhanced immune parameters and the ability of fish to cope with A. hydrophila as a fish pathogen. These findings indicated the potential of these substances as a promising feed additive.

Calcium oxide nanoparticles ameliorate cadmium toxicity in alfalfa seedlings by depriving its bioaccumulation, enhancing photosystem II functionality and antioxidant gene expression

Cadmium (Cd) is a highly toxic and carcinogenic pollutant that poses significant risks to living organisms and the environment, as it is absorbed by the plant roots and accumulates in different parts of crop during its production. A promising sustainable strategy to counteract these threats to use calcium oxide nanoparticles (CaO-NPs) as soil supplements in fodder crops. This approach has shown notable morpho-physiological and biochemical improvements under metal toxicity conditions. However, the specific mechanisms driving Cd tolerance, particularly at physio-biochemical level and antioxidant related genes expression in fodder crops including alfalfa remain unexplored. CaO-NPs supplementation can trigger various signaling pathways that lead to enhance the photosynthetic pigments formation, stomatal conductance, CO2 assimilation rate and quantum yield of photosystem II. In this study, we evaluated various doses of CaO-NPs (0, 25, 50, and 100 mg kg−1) for their efficacy in reducing Cd bioavailability and toxicity in alfalfa plants. Our results demonstrated that Ca2+ and Cd2+, which share the same ionic radius, compete for ion transport through channels. The small size and high availability of CaO-NPs facilitate their rapid translocation within plant tissues, reducing metal uptake by 61 % in shoots and 30 % in roots. Notably, application of CaO-NPs at 100 mg kg−1 significantly increased shoot length (44 %) and root lengths (35 %) as compared to Cd-treated control plants. The highest dose of CaO-NPs also improved photosynthetic efficiency and gas exchange attributes including (gs, Tr, Pn and Ci) by 66 %, 27 %, 33 % and Ci 21 %, respectively, compared with the Cd treated control. Moreover, CaO-NPs (at 100 mg kg−1) alleviated metal-induced oxidative stress by boosting antioxidant enzyme activity like SOD (25 %) POD (42 %), CAT (72 %) and APX (87 %) and diminishing reactive oxygen species (ROS) production when compared with sole Cd treatment. Scanning and transmission electron microscopy revealed that CaO-NPs positively impacted stomatal conductance and mitigated Cd toxicity in leaf ultrastructure. Additionally, the highest dose of CaO-NPs markedly upregulated the expression of antioxidant-related genes, MsCu/Zn SOD, MtPOD, MtCAT, and MtAPX in roots and shoots by 0.67 and 1.03 fold-change (FC), 0.61 and 0.53 FC, 0.54 and 0.88 FC, and 0.46 and 0.66 FC, respectively. In conclusion, CaO-NPs demonstrate significant potential for environmentally friendly mitigation of Cd stress in alfalfa by reducing its uptake, thereby supporting sustainable agriculture.

Isoliquiritigenin as a modulator of the Nrf2 signaling pathway: potential therapeutic implications.

Nuclear factor erythroid-2-related factor 2 (Nrf2), a transcription factor responsible for cytoprotection, plays a crucial role in regulating the expression of numerous antioxidant genes, thereby reducing reactive oxygen species (ROS) levels and safeguarding cells against oxidative stress. Extensive research has demonstrated the involvement of Nrf2 in various diseases, prompting the exploration of Nrf2 activation as a potential therapeutic approach for a variety of diseases. Consequently, there has been a surge of interest in investigating the Nrf2 signaling pathway and developing compounds that can modulate its activity. Isoliquiritigenin (ISL) (PubChem CID:638278) exhibits a diverse range of pharmacological activities, including antioxidant, anticancer, and anti-tumor properties. Notably, its robust antioxidant activity has garnered significant attention. Furthermore, ISL has been found to possess therapeutic effects on various diseases, such as diabetes, cardiovascular diseases, kidney diseases, and cancer, through the activation of the Nrf2 pathway. This review aims to evaluate the potential of ISL in modulating the Nrf2 signaling pathway and summarize the role of ISL in diverse diseases prevention and treatment through modulating the Nrf2 signaling pathway.

Immunomodulatory and growth promotors effects of rosemary (Rosmarinus officinalis) and/or Spirulina with respect to some gene expression on Nile tilapia (Oreochromis niloticus) in Aswan Governorate

The purpose of this work trial was to determine the true impact of spirulina and rosemary in a combination treatment on Nile tilapia performance before experimental infection. Moreover, the effects of these treatments on the liver and kidney functions, the histology of certain organs, and the expression of certain antioxidant and immune-related genes in the liver tissue both before and after Aeromonas hydrophila (A. hydrophila) infection are also included. To achieve this purpose, four diets were prepared: the first was free of additives as a control group (Con), the second contained (SP) spirulina at 7.5 g/kg of diet, the third contained (RM) rosemary at a rate of 10 g/kg of diet, and the fourth one contained rosemary and spirulina as a SP + RM group. The results of the different parameters indicated the growth-stimulant effects of rosemary and spirulina either alone or in a combination when compared with the control group (P ≤ 0.05), with a superior effect to combination treatment. In addition, improved hematological parameters, immunological assays, phagocytic activity and index, and expression of antioxidant and immune-related genes occurred before and after infection. Moreover, there were positive effects of spirulina and rosemary without pathological lesions in the different organs before infection and fewer lesions after infection.

Synthesis, characterization and MAFLD prevention potential of Ganoderma lucidum spore polysaccharide-stabilized selenium nanoparticles

The unstability of selenium nanoparticles (SeNPs) results in decreased activity which limits its therapeutic potential. In this study, we utilized Ganoderma lucidum spore polysaccharide (GLP, Mw = 983.96 kDa) as a novel stabilizer to synthesize GLP-SeNPs. GLP-SeNPs (Se/GLP = 1/3) with an average diameter of 149 nm were successfully prepared and it was stable for at least 30 days at 4 °C. It exhibited an orange-red color, zero valence state, amorphous structure, selenium uniform distribution, a zeta potential of −29.73 mV, selenium content of 16.04 %. GLP-SeNPs pretreatment decreased lipid accumulation, reduced ROS content and enhanced SOD and CAT activity in HepG2 cells. Fe2+ and MDA contents were decreased, while GPX4 and GSH activities were increased. All these ameliorated effects could be abolished by NRF2 antagonist ML385. The expression of anti-oxidant genes and iron exporter was up-regulated, while that of pro-oxidant and lipid biosynthesis gene was down-regulated. The GPX4 activity could be reduced by ML385 addition. In conclusion, GLP-SeNPs was successfully constructed at the ratio of 1/3 (Se/GLP). It prevents MAFLD by targeting ferroptosis, including lowering iron overload, inhibiting lipid accumulation and attenuating oxidative stress. The improvement was conducted via activating SLC40A1-mediated iron pathway, ACSL4-mediated lipid metabolism and NRF2-mediated GSH-GPX4 pathway. Therefore, GLP-SeNPs can be used as potential selenium nutritional supplements or adjuvants for MAFLD prevention.

Pearl Millet (Pennisetum glaucum): A Climate Resilient and Nutritionally Significant Crop for Global Food Security

The environmental balance needed for agriculture is severely strained due to changes in rainfall patterns and climate change, as well as the growing global need for food. As global temperatures rise and precipitation patterns become more erratic, millets offer an efficient solution to the trade-off between food security and water availability. Millets are rich in nutrients and possess a special ability to endure extreme conditions such as heat, drought, and various other abiotic and biotic stresses. Pearl millet is a highly climate-resilient crop capable of mitigating the adverse impacts of climate change. Antioxidant gene expression, presence of a waxy outer layer and heat shock proteins/chaperons contribute to its abiotic stress tolerance. By adjusting its flowering phenology and establishing fast-growing roots, pearl millet enhances its water and nutrient use efficiency. Stable sources of germplasm in pearl millet that exhibit resistance to diseases such as downy mildew, blast, rust, smut, and ergot have also been incorporated into disease resistance breeding programs. Pearl millet plays a crucial role in nourishing the poverty-stricken populations due to its high palatability and rich nutritional profile. Various bioactive compounds pertain to hypoglycemic, anticancerous and probiotic properties that contribute to its health benefits. Biofortification in pearl millet offers a solution to address issues of micronutrient deficiency in vulnerable populations. Continued research and investment in pearl millet production not only bolster food security but also contribute to the resilience of agricultural systems in the face of increasingly unpredictable environmental conditions. Thus, embracing pearl millet represents a forward-thinking approach to ensuring food security and promoting nutritional diversity on a global scale.

Assessing the impact of weathered polystyrene collected from the marine environment on oxidative stress responses in Zophobas morio larvae: A preliminary study

Foamed polystyrene (PS) is a prevalent material in consumer products and thus represents one of the largest constituents of marine litter. PS may interact with and be ingested by various organisms, including insects, which are starting to be used in biodegradation of plastic wastes. This study examines the physiological performances and the potential oxidative stress related to dietary environmental PS exposure of Z. morio larvae. Experimental groups were fed different diets: bran and oatmeal (control), weathered PS collected from the marine environment, and virgin PS. Over a 30-day feeding period, larvae growth, survival, and PS consumption were measured, together with antioxidant enzymatic activities and gene expression profiles. Results showed that PS ingestion supports larval growth similarly to bran, but the two PS-fed groups consumed a different amount of plastic, suggesting that weathered PS might affect the patterns of PS consumption by larvae. We denoted that PS consumption was associated with a decline in total antioxidant capacity, which showed the highest decrease in the environmental PS group. Marked increases in oxidative stress biomarkers such as superoxide dismutase (SOD) activity/mRNA levels and malondialdehyde content were also observed in the environmental PS-fed group, indicating an elevated oxidative stress response, probably due to pollutants adsorbed/absorbed on PS. Differently to SOD, catalase at both mRNA and enzyme activity levels was found to be significantly reduced by PS digestion, regardless of PS type. The mRNA levels of glutathione s-transferase (GST) were significantly higher in the environmental PS-fed larvae compared to the virgin PS-fed group. In addition, Principal Component Analysis clearly distinguished between larvae fed different PS types, highlighting the enhanced oxidative stress caused by the ingestion of PS collected from marine environments. These results support the potential of insects in plastic biodegradation processes but also demonstrate the health hazards of using environmental PS as principal dietary component in Z. morio larvae, thus questioning their use in downstream productions.

Dietary Artichoke (Cynara scolymus) Extract Ameliorated the Growth Performance, Humoral Immune Parameters and Resistance Against Aeromonas hydrophila in Goldfish (Carassius auratus)

Abstract This trial investigated the efficacy of artichoke (Cynara scolymus) extract (AE) on the growth performance, immunity, antioxidant parameters, and resistance against Aeromonas hydrophila of goldfish (Carassius auratus). For this purpose, a total number of 470 goldfish with initial weight 5.70±0.2 g were fed with four experimental diets including 0 (T0), 100 (T1), 150 (T2), and 200 (T3) mg kg−1 diet AE for 8 weeks. At the end of feeding trial, growth performances, serum immune parameters, and mucus antioxidant enzymes were measured. Fish were challenged with A. hydrophila, and the antioxidant and immunity-related gene expression were investigated. Based on the results, the highest final weight (FW) and weight gain (WG) were attained in T2 and T3 (P<0.05). Immune factors including ACH50, lysozyme, and total immunoglobulin in T2 and T3 showed the highest values (P<0.05). The expression of GR, IL1 β, TNF α, HSC70, HSP70, and HSP90 β genes in T1, T2, and T3 were higher than the control (P<0.05). The GST expression was significantly enhanced in T2 (P<0.05). The present study demonstrated that the administration of AE, especially at doses of 150 mg kg−1, could improve the growth, immunity, and antioxidant parameters, as well as enhance disease resistance against A. hydrophila in goldfish.

Whole-transcriptome analysis reveals the effect of retinoic acid on small intestinal mucosal injury in cage-stressed young laying ducks

Retinoic acid (RA) is an active derivative of vitamin A and is involved in a variety of physiological processes, including cell growth, antioxidant, and inflammation. However, the role of RA in intestinal oxidative stress injury in caged-stressed laying ducks is unknown. In this study, we analyzed the effect and underlying mechanism of RA supplementation on intestinal damage in cage-stressed young laying ducks. One hundred and sixty laying ducks were divided into 5 treatment groups, including a control group (CR) and 4 treatment groups exposed to different RA concentrations (2,500, 5,000, 7,500 and 10,000 IU/kg, TG1 to TG4). The experimental period comprised a 7-d prefeeding period and a 10-d experimental feeding period, for a total of 17 d. Phenotypic analysis revealed that compared with the control group, RA addition increased the intestinal villus height and the villus-to-crypt ratio; decreased the crypt depth (P < 0.01); decreased the serum diamine oxidase and D-lactate concentrations (P < 0.05); increased the serum antioxidant capacity and intestinal antioxidant gene expression levels (P < 0.05); and increased the expression levels of tight junction-related genes, with the greatest effect observed in TG2 group. Our further whole-transcriptome analysis of duodenum tissues from CR and TG2 ducks revealed 706 differentially expressed mRNAs (DEmRNAs), 357 differentially expressed lncRNAs (DElncRNAs), 14 differentially expressed circRNAs (DEcircRNAs), and 4 differentially expressed miRNAs (DEmiRNAs). These DEGs are involved in calcium signaling, NOD-like receptor signaling, pyruvate metabolism, Jak-STAT signaling, Wnt signaling, riboflavin metabolism, and the adherens junction and tight junction pathways. The results of omics and marker gene expression analysis suggested that RA treatment may play a role in endoplasmic reticulum stress (ERS) and apoptosis. In conclusion, the addition of RA to the diet improved intestinal injury by improving the redox homeostasis of intestinal cells associated with ERS, enhancing the intestinal tight junction structure and alleviating the apoptosis of intestinal epithelial cells; moreover, 5,000 IU/kg RA was determined to be the most appropriate concentration for supplementation.

Bisphenol S decreased lifespan and healthspan via insulin/IGF-1-like signaling-against mitochondrial stress in Caenorhabditis elegans

Bisphenol S (BPS) is widely presented and affects aging with unclear mechanisms. Here, we applied C. elegans to evaluate the effects of BPS on lifespan and healthspan and to investigate the underlying mechanisms. Both early-life and whole-life exposure to BPS at environmentally relevant doses (0.6, 6, 60 μg/L) significantly decreased lifespan, and healthspan (body bend, pharyngeal pumping, and lipofuscin accumulation). BPS exposure impaired mitochondrial structure and function, which promoted ROS production to induce oxidative stress. Furthermore, BPS increased expressions of the insulin/IGF-like signaling (IIS). Also, BPS inhibited expression of the IIS transcription factor daf-16 and its downstream anti-oxidative genes. Quercetin effectively improved BPS-induced oxidative stress byreversing BPS-regulated IIS/daf-16 pathway and anti-oxidative gene expressions. In daf-2 and daf-16 mutants, the effects of BPS and quercetin on lifespan, healthspan, oxidative stress, and anti-oxidative genes expressions were reversed, demonstrating the requirement of IIS/daf-16 for aging regulation. Molecular docking and molecular dynamics simulations confirmed the stable interaction between DAF-2 and BPS mainly via three residues (VAL1260, GLU1329, and MET1395), which was attenuated by quercetin. Our results highlighted that adverse effects of BPS on impairing lifespan and healthspan by affecting IIS/daf-16 function against mitochondrial stress, which could be inhibited by quercetin treatment. Thus, we first revealed the underlying mechanisms of BPS-induced aging and the potential treatment.

Effects of passion fruit peel (Passiflora edulis) pectin and red yeast (Sporodiobolus pararoseus) cells on growth, immunity, intestinal morphology, gene expression, and gut microbiota in Nile tilapia (Oreochromis niloticus)

This study explores the effects of dietary supplementation with passion fruit peel pectin (Passiflora edulis) and red yeast cell walls (Sporidiobolus pararoseus) on growth performance, immunity, intestinal morphology, gene expression, and gut microbiota of Nile tilapia (Oreochromis niloticus). Nile tilapia with an initial body weight of approximately 15 ± 0.06 g were fed four isonitrogenous (29.09–29.94%), isolipidic (3.01–4.28%), and isoenergetic (4119–4214 Cal/g) diets containing 0 g kg−1 pectin or red yeast cell walls (T1 - Control), 10 g kg−1 pectin (T2), 10 g kg−1 red yeast (T3), and a combination of 10 g kg−1 pectin and 10 g kg−1 red yeast (T4) for 8 weeks. Growth rates and immune responses were assessed at 4 and 8 weeks, while histology, relative immune and antioxidant gene expression, and gut microbiota analysis were conducted after 8 weeks of feeding. The results showed that the combined supplementation (T4) significantly enhanced growth performance metrics, including final weight, weight gain, specific growth rate, and feed conversion ratio, particularly by week 8, compared to T1, T2, and T3 (P < 0.05). Immunological assessments revealed increased lysozyme and peroxidase activities in both skin mucus and serum, with the T4 group showing the most pronounced improvements. Additionally, antioxidant and immune-related gene expression, including glutathione peroxidase (GPX), glutathione reductase (GSR), and interleukin-1 (IL1), were upregulated in the gut, while intestinal morphology exhibited improved villus height and width. Gut microbiota analysis indicated increased alpha and beta diversity, with a notable rise in beneficial phyla such as Actinobacteriota and Firmicutes in the supplemented groups. These findings suggest that the combined use of pectin and red yeast cell walls as prebiotics in aquaculture can enhance the health and growth of Nile tilapia, offering a promising alternative to traditional practices. Further research is needed to determine optimal dosages for maximizing these benefits.

Ameliorative Effect of Hedysarum polybotrys Polysaccharide on Neural Tissue Fibrosis in Diabetic Peripheral Neuropathy Mice and its Mechanisms

Objective: This study aimed to investigate the role of Hedysarum polybotrys polysaccharide (HPS) in ameliorating neural tissue fibrosis in diabetic peripheral neuropathy (DPN) mice. Methods: Fifty DPN mice were selected and randomly divided into five groups (n = 10), which were the model group, positive control group (receiving only 4 mg/(kg-d) of α-lipoic acid), high-dose HPS group (200 mg/(kg-d) of HPS was given per day), medium-dose HPS group (100 mg/(kg-d) of HPS was given per day), and low-dose HPS group (50 mg/(kg-d) of HPS given daily). In addition, non-diabetic C57BL/6 wild-type mice were selected as the normal group (n = 10). The expression levels of Keap1 and Nrf2 proteins and their mRNAs in the sciatic nerve tissues of mice in each group were analyzed by Western blot technique and real-time fluorescence quantitative PCR. Results: Compared with the normal group, the expression of Keap1 protein and mRNA was increased, while the expression of Nrf2 protein and mRNA was decreased in the sciatic nerve of mice in the model group (P &lt; 0.05). Compared with the model group, Keap1 protein and mRNA expression decreased, while Nrf2 protein and mRNA expression increased in the control and high and medium dose HPS groups of mice (P &lt; 0.05). Conclusion: HPS may inhibit fibrosis of neural tissue and ameliorate nerve injury in DPN mice by regulating the Keap1/Nrf2 signaling pathway. This effect was associated with enhanced antioxidant capacity, promotion of Nrf2 activation, and increased antioxidant gene expression by HPS. Therefore, HPS has a potential therapeutic value to ameliorate DPN-associated nerve injury.

Integrating network pharmacology and experimental verification to study the mechanism of Polygonum hydropiper total flavonoids against stress-induced gastric mucosal damage

ContextPolygonum hydropiper L. (P. hydropiper) has outstanding clinical efficacy in treating both acute and chronic gastroenteritis. However, the definite mechanism remains unclear. ObjectiveThis study aimed to explore the potential mechanisms of the total flavonoid of P. hydropiper (FPH) in stress-induced gastric mucosal damage (SGMD) rats through a combination of network pharmacology, molecular docking, and animal experiments. MethodsNetwork pharmacology and molecular docking were utilized to predict the potential mechanisms of FPH against SGMD. In experimental studies, SGMD rat models were established using water-immersion restraint stress (WIRS). FPH was administered at doses of 140, 70, and 35 mg/kg, with ranitidine serving as a positive control, through gavage once daily for 6 consecutive days after model establishment. Stomach and serum specimens were analyzed using HE staining, Western blotting, qPCR, and ELISA to investigate the protective mechanism of FPH in SGMD. ResultsThe network pharmacology analysis identified 16 active ingredients and 183 common targets, with potential pathways including PI3K/Akt, MAPK and Keap1/Nrf2. In vivo experiments demonstrated that FPH intervention alleviated SGMD pathological changes, reduced elevated serum IL-6 and TNF-α levels, and enhanced SOD and GSH activity in rats. Additionally, FPH increased the protein expression of p62, Nrf2, HO-1, PI3K, and p-Akt, along with mRNA levels of Nrf2, p62, and HO-1. ConclusionsFPH exerts a gastric mucosal protective effect by upregulating antioxidant gene expression through the PI3K/Akt and Keap1/Nrf2 pathways. This study provides an experimental basis for the potential clinical treatment of SGMD with the traditional Chinese medicine P. hydropiper.

NQO1 from Litopenaeus vannamei involved in the regulation of antioxidant capacity, inflammation, and apoptosis

NAD(P)H:quinone oxidoreductase 1 (NQO1) is a versatile FAD-dependent flavoprotein and a key regulator in the Nrf2/ARE signaling pathway. In this study, we cloned the NQO1 gene of Litopenaeus vannamei (LvNQO1) and investigated its role in immune regulation. The results showed that the LvNQO1 gene has an ORF of 987 bp, encoding 328 amino acids. The protein contains an NQO family domain. Homology and phylogenetic analysis revealed that LvNQO1 shares similarities with the NQO1 of Marsupenaeus japonicus, clustering into the same clade. LvNQO1 was expressed in 10 tissues of L. vannamei, with the highest expression in the hepatopancreas and the lowest in the intestine. After RNA interference (RNAi) of LvNQO1, the expression of antioxidant genes (SOD and GST) initially increased but then decreased after 24 h. The expression levels of Gpx and CAT decreased early on but gradually recovered. Immune-related factors (Toll4, JNK, and Hsp90) and apoptosis-related factors (Caspase3, P53, and bcl2) were upregulated. Following RNAi of LvNQO1, Vibrio harveyi challenge significantly increased the expression of immune factors (JNK and HIF1α) and apoptosis factors (Caspase3, P53, and bcl2) in the hepatopancreas. Toll 4 receptors recognized the infection, upregulated their expression, and activated an immune response. RNAi of LvNQO1 led to noticeable histological lesions and apoptosis. These findings suggest that LvNQO1 may play a role in regulating antioxidant capacity, inflammation, and apoptosis in response to Vibrio infection.

Self-cascade ROS-trapping bioreaction system reverses stem cell oxidative stress fate for osteogenesis

eactive oxygen species (ROS) scavenging is essential for periodontal regeneration. However, the dynamic change of the applied materials within the ROS-rich environment and the residual oxidation products in the host highly impact periodontal regeneration. This study successfully constructs a bioreaction system via thiol-ene click chemistry, leveraging the high affinity of glutathione (GSH) for ROS to attract excess ROS to the crosslinking points. Two minutes after hydrogen peroxide (H2O2) treatment, the ROS level in the G8-0 hydrogel acutely decreases, reaching a 4.4% reduction within 10minutes, confirming the ROS-trapping efficacy. Through a ‘bait switch-on’ mechanism, hexagonal boron nitride (hBN) takes over the captured ROS and the oxidation products of pectin further drive the reduction reaction, ultimately restoring the extracellular environment. The self-cascade products, oxidized hBN, reshape the intracellular oxidative stress (OS) environment, achieving a synergistic extra- and intra-cellular treatment. The significantly high reduced to oxidized glutathione (GSH/GSSG) ratio in G8-10 hydrogel (~80%) illustrates a reversal of oxidative stress in bone marrow stem cells (BMSCs). On a molecular level, the bioreaction system inhibits the NF-κB pathway, promoting the expression of key antioxidant genes (Nqo1 and Nrf2) and osteogenic molecules (ALP and OCN), thereby reversing the detrimental effects of OS on BMSCs. In vivo application demonstrated the system’s strong redox-balancing and osteogenic capabilities in the periodontal inflammation environment. This novel antioxidant bioreaction system, characterized by self-cascade ROS-trapping and product utilization, offers innovative treatment strategies for tissue regeneration under conditions of excessive OS.

NRF2 protects lung epithelial cells from wood smoke particle toxicity

Wildfire smoke is a potential source of oxidative stress in lung epithelial tissue. The response to oxidative stress is controlled by the transcription factor NRF2, which is the central regulator of antioxidant gene expression. If wood smoke particle (WSP) exposure induces reactive oxygen species (ROS) in epithelial cells, then NRF2 may protect against pathological conditions resulting from increased oxidative stress through changes in gene expression. We used two lung epithelial cell lines to test this hypothesis in vitro: A549, which harbor a mutation resulting in constitutive activation of NRF2, and BEAS-2B, which show limited NRF2 activity under basal conditions, but high inducibility during oxidative stress. In BEAS-2B cells, WSP exposure leads to increased cellular ROS, activation of NRF2, and upregulation of the NRF2 target genes NQO1, GCLM, and SRXN1. WSP exposure also increased ROS in A549 cells, although NRF2 activation and antioxidant gene upregulation were less robust as both were basally high in this cell line. Overall, the degree of ROS induction by WSP across cell lines is dependent upon NRF2 activity, and a similar pattern was observed for WSP cytotoxicity. WSP also sensitized both cell lines to the ferroptosis inducer erastin in a manner that is correlated with NRF2 activity. Knockout of NRF2 in A549 resulted in higher WSP-induced ROS generation, cytotoxicity, and erastin sensitivity. Taken together, these results suggest that NRF2 serves as a protective factor against wood smoke induced ROS and oxidative stress in lung epithelial cells.

Specific cancer types and prognosis in patients with variations in the KEAP1-NRF2 system: A retrospective cohort study.

The KEAP1-NRF2 system induces the expression of antioxidant genes in response to various types of oxidative stress. Some cancer cells activate this system, which increases their malignancy through genetic mutations. We performed a retrospective cohort study using the C-CAT database, which contains the gene-panel sequence data from 60,056 cases of diagnosed solid tumors. We analyzed somatic mutations in NRF2 and KEAP1 genes and their associations with clinical outcomes. Variants in the NRF2 gene were clustered in exon 2, which encodes the DLG and ETGE motifs essential for KEAP1 interaction. The NRF2 variants were frequently observed in esophageal and lung squamous cell carcinoma with frequencies of 35.9% and 19.6%, respectively. Among these mutations, the NRF2 variants in the ETGE motif were indicators of a worse prognosis. KEAP1 variants were found in 2.5% of all cases. The variants were frequent in lung cancer and showed a worse prognosis in lung and other types of adenocarcinomas. We then conducted gene expression analysis using TCGA data. While cancers with DLG and ETGE variants were similar in terms of gene expression profiles, there were significant differences between cancers with KEAP1 and NRF2 variants. Our results indicate that genetic alteration of the KEAP1-NRF2 pathway is a major factor in patient prognosis for each cancer type and its genetic variant. Variants in NRF2 and KEAP1 genes can characterize the biological basis of each cancer type and are involved in carcinogenesis, resistance to therapy, and other biological differences.