Expression Of Antioxidant-related Genes Research Articles

A blend of chitosan-vitamin C and vitamin E nanoparticles robust the immunosuppressed- status in Nile tilapia treated with salt

In aquaculture, fish are exposed to many stressors, such as climate changes and infectious diseases that affect their performance, immunity, and welfare. Freshwater fish subjected to salt bath become exhausted and stressed. In this experiment, Nile tilapia were exposed to a salt bath at a dose of 30 ppt for 30 min a day. Vitamin C and vitamin E are well-known antioxidants that are used in aquaculture. Fish received dietary nanoparticles of chitosan-vitamin C and chitosan-vitamin E (CCE-NPs) for different periods (7 and 14 days) pre- (G2) and post-salt treatment (G3). In the control fish (G1), cortisol 5.44 µg/dL and glucose 91.67 mg/dL were significantly up-regulated post-salt treatment by 1 h and 24 h, respectively, whereas those (G2) fed CCE-NPs diet had significantly lower values of 4.72 and 3.25 µg/dL; 86.3 and 84.3 mg/dL, respectively. A rapid decrease of glucose 68.3 and 66.3 mg/dL was noticed in those (G2) fed CCE-NPs diet compared to the control 84.67 mg/dL at 48 h post-stress. Regardless of the supplementation period, fish (G2) could partially restore normal food reflex at 48 h (post-salt bath) and fully restored at 72 h compared to 7 days in the control (G1). After 48 h, fish that received dietary CCE-NPs (G2 and G3) restored normal mucus lysozyme levels, whereas the control did not restore pre-treatment values till the seventh day. Mucus antibacterial activity, fish received rapid dietary CCE-NPs (G2) and partially restored average values (pre-salt bath) at 96 h. The salt treatment could provoke gene expression of pro-inflammatory cytokines interleukin (IL-1β) and tumor necrosis (TNF)-α in the head kidney of fish at 24 h post-salt bath to 5.9–8.35 fold-change, respectively, with a rapid decline in fish (G2) the gene expression. Post-salt bath (24 h), the gene expression of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) was higher in fish (G2) than in the control group (G1) regardless of the supplementation period (7 and 14 days). Bacterial infection S. agalactiae (OL471408), a significantly lower MR was recorded in G2 at 40% and 33.3% compared to the control G1 MR (53.3%), with an RPL of 24.95% and 37.5%. In conclusion, Nile tilapia treated with a 30 ppt salt became more vulnerable to S. agalactiae. Adding CCE-NPs to the Nile tilapia diet for 7- and 14-day pre-salt bath could increase immune and antioxidant-related gene expression to counteract S. agalactiae infection.

Effects of Different Salinity Stress on the Transcriptomic Responses of Freshwater Crayfish (Procambarus clarkii, Girard, 1852)

Salinization of freshwater ecosystems is a pressing global issue. Changes in salinity can exert severe pressure on aquatic animals and jeopardize their survival. Procambarus clarkii is a valuable freshwater aquaculture species that exhibits some degree of salinity tolerance, making it an excellent research model for freshwater aquaculture species facing salinity stress. In the present study, crayfish were exposed to acute low salt (6 ppt) and high salt (18 ppt) conditions. The organisms were continuously monitored at 6, 24, and 72 h using RNA-Seq to investigate the mechanisms of salt stress resistance. Transcriptome analysis revealed that the crayfish responded to salinity stress with numerous differentially expressed genes, and most of different expression genes was observed in high salinity group for 24h. GO and KEGG enrichment analyses indicated that metabolic pathways were the primary response pathways in crayfish under salinity stress. This suggests that crayfish may use metabolic pathways to compensate for energy loss caused by osmotic stress. Furthermore, gene expression analysis revealed the differential expression of immune and antioxidant-related pathway genes under salinity stress, implying that salinity stress induces immune disorders in crayfish. More genes related to cell proliferation, differentiation, and apoptosis, such as the Foxo, Wnt, Hippo, and Notch signaling pathways, responded to high-salinity stress. This suggests that regulating the cellular replication cycle and accelerating apoptosis may be necessary for crayfish to cope with high-salinity stress. Additionally, we identified 36 solute carrier family (SLC) genes related to ion transport, depicting possible ion exchange mechanisms in crayfish under salinity stress. These findings aimed to establish a foundation for understanding crustacean responses to salinity stress and their osmoregulatory mechanisms.

PrG Protects Postovulatory Oocytes Aging in Mice through the Putrescine Pathway

Postovulatory aging of oocytes involves a series of deleterious molecular and cellular changes, which adversely affect oocyte maturation, fertilization, and early embryonic development. Petunidin-3-O-(6-O-pcoumaroyl)-rutinoside-5-O-glucoside (PrG), the main active ingredient of anthocyanin, exerts antioxidant effects. This study investigated whether PrG supplementation could delay postovulatory oocyte aging by alleviating oxidative stress. Our results showed that PrG supplementation decreased the number of abnormal morphology oocytes and improved the oxidative stress of aged oocytes by facilitating the reduction of the reactive oxygen species, the increase in glutathione content, and the recovery of expression of antioxidant-related gene expression. In addition, PrG treatment recovered mitochondrial dysfunction, including mitochondrial distribution, mitochondrial membrane potential and adenosine triphosphate in aged oocytes. PrG-treated oocytes returned to normal levels of cytoplasmic and mitochondrial calcium. Notably, PrG inhibited early apoptosis in aged oocytes. RNA-seq and qRT-PCR results revealed that PrG ameliorated oxidative stress injury in postovulatory aging oocytes of mice via the putrescine pathway. In conclusion, in vitro PrG supplementation is a potential therapy for delaying postovulatory oocyte aging.

GABA attenuates neurotoxicity of zinc oxide nanoparticles due to oxidative stress via DAF-16/FoxO and SKN-1/Nrf2 pathways

Zinc oxide nanoparticles (ZnO-NPs) are one of the most widely used metal oxide nanomaterials. The increased use of ZnO-NPs has exacerbated environmental pollution and raised the risk of neurological disorders in organisms through food chains, and it is urgent to look for detoxification strategies. γ-Aminobutyric acid (GABA) is an inhibitory neurotransmitter that has been shown to have anxiolytic, anti-aging and inhibitory effects on nervous system excitability. However, there are few reports on the prevention and control of the toxicity of nano-metal ions by GABA. In zebrafish, ZnO-NPs exposure led to increased mortality and behavioral abnormalities of larva, which could be moderated by GABA intervention. Similar results were investigated in Caenorhabditis elegans, showing lifespan extension, abnormal locomotor frequency and behavior recovery when worms fed with GABA under ZnO-NPs exposure. Moreover, GABA enhanced antioxidant enzyme activities by upregulating the expression of antioxidant-related genes and thus scavenged excessive O2−. In the case of ZnO-NPs exposure, inhibition of nuclear translocation of DAF-16 and SKN-1 was restored by GABA. Meanwhile, the protective effect of GABA was blocked in daf-16 (−) and skn-1 (−) mutant, suggesting that DAF-16/FoxO and SKN-1/Nrf2 pathways is the key targets of GABA. This study provides a new solution for the application of GABA and mitigation of metal nanoparticle neurotoxicity.

Zinc oxide application alleviates arsenic-mediated oxidative stress via physio-biochemical mechanism in rice.

Arsenic (As) pollution in cultivated soils poses a significant risk to the sustainable growth of agriculture and jeopardizes food security. However, the mechanisms underlying how zinc (Zn) regulates the toxic effects induced by As in plants remain poorly understood. Hence, this study aimed to explore the potential of ZnO as an effective and environmentally friendly amendment to alleviate As toxicity in rice, thereby addressing the significant risk posed by As pollution in cultivated soils. Through a hydroponic experiment, the study assessed the mitigating effects of different ZnO dosages (Zn5, 5mg L-1; Zn15, 15mg L-1; Zn30, 30mg L-1) on rice seedlings exposed to varying levels of As stress (As0, 0µM L-1; As25, 25µM L-1). The findings of the study demonstrate significant improvements in plant height and biomass (shoot and root), with a notable increase of 16-40% observed in the Zn15 treatment, and an even more substantial enhancement of 29-53% observed in the Zn30 treatment under As stress, compared to respective control treatment. Furthermore, in the Zn30 treatment, the shoot and root As contents substantially reduced by 47% and 63%, respectively, relative to the control treatment. The elevated Zn contents in shoots and roots enhanced antioxidant enzyme activities (POD, SOD, and CAT), and decreased MDA contents (13-25%) and H2O2 contents (11-27%), indicating the mitigation of oxidative stress. Moreover, the expression of antioxidant-related genes, OsSOD-Cu/Zn, OsCATA, OsCATB, and OsAPX1 was reduced when rice seedlings were exposed to As stress and significantly enhanced after Zn addition. Overall, the research suggests that ZnO application could effectively mitigate As uptake and toxicity in rice plants cultivated in As-contaminated soils, offering potential solutions for sustainable agriculture and food security.

In vivo dynamics of hydrogen peroxide and regulation of antioxidant gene expression in mouse skeletal muscle under cooling stimulus

Introduction: The dynamics of reactive oxygen species (ROS) in the cytosol and mitochondria function as signaling factors that regulates diverse cellular adaptations. Hydrogen peroxide (H2O2) is considered one of the most influential ROS in regulating physiological function; being a ubiquitous molecule with a long half-life and high diffusing capacity compared to other ROS (e.g., superoxide and hydroxyl radicals). Temperature changes within skeletal muscle myocytes constitute an environmental factor that modulates the expression of antioxidant-related genes. Although muscle temperature reduction by icing is a typical physical therapy modality, the relationship between cooling stimuli and in vivo H2O2 dynamics remains unclear. The purpose of this investigation was to elucidate the H2O2 dynamics in the cytosol and mitochondria induced by cooling stimuli and to determine the relationship between these dynamics and the expression of antioxidant-related genes. Specifically, we tested the hypothesis that cooling would reduce [H2O2] in intracellular compartments (cytosol and mitochondria) and suppress antioxidant-related gene expression levels. Methods: Experiment 1. in vivo H2O2 dynamics observation: Cytosolic and mitochondrial H2O2-sensitive fluorescent proteins (HyPer7, n = 6 and MLS-HyPer7, n = 9) were expressed by electroporation in the anterior tibialis muscle (TA) of male C57/BL6 mice (9-14 week of age). The TA was exposed under anesthesia, and cytosolic and mitochondrial [H2O2] were measured when the muscle temperature was reduced from 35 °C to 0 °C using in vivo bioimaging techniques. Experiment 2. Mice (9-12 week of age) were divided into an icing (ICE) group that received cooling stimulation (muscle temperature reduced from 35 °C to 13 °C and back to 35°C, total 6 sets for 60 min, n = 6) and a control (CONT) group (muscle temperature 35 °C for 60 min, n = 6). At 3 hours after temperature intervention, the TA was removed and mRNA levels were measured by Real-time PCR. Results: Experiment 1. In vivo [H2O2] in each compartment were significantly lower in the cytosol (over the range 28-6°C) and mitochondria (over the range 29-1°C) compared to 35 °C conditions. A decreasing phase of [H2O2] in both compartments was observed from 35 °C to around 13 °C, but, subsequently increasing [H2O2] were observed in the lower temperature range (i.e., 12 to 0 °C). Experiment 2: The ICE group significantly increased mRNA levels of antioxidant enzyme-related genes Nfe2l2 and HSF1 compared to CONT (1.2 ± 0.1-fold and 1.1 ± 0.1-fold, respectively, P < 0.01). On the other hand, mRNA levels of mitochondria-related genes such as PGC1α and COX4 were unchanged. Conclusions: This investigation is the first to reveal the H2O2 and related gene expression impact of cooling stimuli in an in vivo model. Cooling mouse skeletal muscle produced temperature-dependent decreases in [H2O2] that reversed with more severe cooling. In both cytosolic and mitochondrial compartments, the lowest point for [H2O2] was found to be 13°C. Repeated cooling cycles (between 35 and 13 °C) with their attendant decreasing [H2O2] elevated antioxidant-related mRNA levels. These results support that icing therapy may improve the antioxidant capacity of muscle despite decreasing muscle ROS levels. This study was supported in part by a Grant-in-Aid for Japan Society for the Promotion of Science (JSPS) for Grant-in-Aid for JSPS Fellows (no. JP23KJ0967) and JSPS KAKENHI Grant (No. JP20H04074). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Differential effects of oxytetracycline on detoxification and antioxidant defense in the hepatopancreas and intestine of Chinese mitten crab under cadmium stress

This study aims to evaluate the effects of oxytetracycline (OTC) on detoxification and oxidative defense in the hepatopancreas and intestine of Chinese mitten crab (Eriocheir sinensis) under cadmium (Cd) stress. The crab was exposed to 0.6 μM Cd, 0.6 μM OTC, and 0.6 μM Cd plus 0.6 μM OTC for 42 days. Our results showed that in the intestine, OTC alone enhanced protein carboxylation (PC) and malondialdehyde (MDA) contents, which was associated with the increased OTC accumulation. Compared to Cd alone, Cd plus OTC increased Cd and OTC contents, and reduced detoxification (i.e., glutathione (GSH) content, gene expressions of cytochrome P450 (CYP) isoforms, 7-ethoxyresorufin O-deethylase (EROD) activity, mRNA levels and activities of glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST)), and antioxidant defense (i.e., gene expressions and activities of catalase (CAT) and superoxide dismutase (SOD)) in the intestine, leading to the increased in PC and MDA contents, suggesting that OTC had a synergistic effect on Cd-induced oxidative damage. In the hepatopancreas, although OTC alone increased OTC accumulation, it did not affect PC and MDA contents. Compared to Cd alone, Cd plus OTC reduced MDA content, which was closely related to the improvement of detoxification (i.e., GSH content, mRNA levels of CYP isoforms, EROD activity, gene expressions and activities of GPx, GR and GST), and antioxidant defense (gene expressions and activities of CAT and SOD, metallothionein content). Aryl hydrocarbon receptor (AhR) and nuclear factor E2-related factor 2 (Nrf2) transcriptional expressions were positively correlated with most detoxification- and antioxidant-related gene expressions, respectively, indicating that AhR and Nrf2 were involved in the regulation of these gene expressions. Our results unambiguously demonstrated that OTC had tissue-specific effects on Cd-induced toxicological effect in E. sinensis, which contributed to accurately evaluating Cd toxicity modulated by TCs in crab.

Impact of a varied combinatorial mixture of β-1, 3 glucan and fructooligosaccharides on growth performance, metabolism, intestinal morphometry, expression of antioxidant-related genes, immunity, and protection against Vibrio alginolyticus in Red tilapia (Oreochromis niloticus x O. mossambicus)

Impact of a varied combinatorial mixture of β-1, 3 glucan and fructooligosaccharides on growth performance, metabolism, intestinal morphometry, expression of antioxidant-related genes, immunity, and protection against Vibrio alginolyticus in Red tilapia (Oreochromis niloticus x O. mossambicus)

In ovo feeding of α-ketoglutaric acid improves hepatic antioxidant-gene expression, plasma antioxidant activities and decreases body temperature without affecting broiler body weight under cyclic heat stress

The broiler industry is adversely affected by the rise in global temperature. This study investigated the effects of in ovo feeding of α-ketoglutaric acid (AKG) on growth performance, organ weight, plasma metabolite, plasma oxidative stress, rectal temperature (RT), and hepatic mRNA expression of antioxidant-related genes in Arbor Acres broilers subjected to cyclic heat stress (HS). Three hundred fifty fertile eggs during incubation were divided into 5 groups according to AKG concentrations and temperature conditions. After dissolving AKG in distilled water at 0, 0.5, 1.0, and 1.5, 0% AKG was in ovo administered to 2 of the 5 groups whereas the remaining 3 groups received 0.5, 1.0, and 1.5%, respectively. From d 29 to 34 of age, 4 groups of birds received heat stress (HS) at 31°C ± 1°C for 6 h per day while the other group was kept at room temperature (21°C ± 1°C; NT). So, the 5 treatment groups were: 1) 0AKG-NT, where chicks hatched from eggs receiving 0% AKG were reared under thermoneutral conditions. 2) 0AKG-HS, where chicks hatched from eggs receiving 0% AKG were reared under cyclic HS conditions. 3) 0.5AKG-HS, where chicks hatched from eggs receiving 0.5% AKG were reared under cyclic HS conditions. 4) 1.0AKG-HS, where chicks hatched from eggs receiving 1.0% AKG were reared under cyclic HS conditions. 5) 1.5AKG-HS, where chicks hatched from eggs receiving 1.5% AKG were reared under cyclic HS conditions. HS significantly reduced body weight change (ΔBW %) and average daily gain (ADG) without affecting average daily feed intake (ADFI). Feed conversion ratio (FCR) was significantly increased (P = 0.003) in all HS-treated groups. A significant linear decrease in the final RT (P = 0.005) and a change in RT (P = 0.003) were detected with increasing AKG concentration. Total antioxidant capacity (P = 0.029) and antioxidant balance (P = 0.001) in plasma increased linearly with increasing AKG concentration whereas malondialdehyde concentrations were linearly decreased (P = 0.001). Hepatic gene expression of CAT (P = 0.026) and GPX1 (P = 0.001) were dose-dependently upregulated while nicotinamide adenine dinucleotide phosphate oxidase (NOX)1, NOX4, and heat shock protein (HSP)70 were linearly downregulated (P < 0.05). Hence, in ovo injection of AKG was effective in mitigating HS-induced oxidative stress without attenuating the adverse effects on broiler growth.

Antioxidant capacity in broiler chicken enhanced by nucleotides and/or beta-glucan, resulting in increased antioxidant-related gene expression plus enzymes and improved meat quality

Antioxidant capacity in broiler chicken enhanced by nucleotides and/or beta-glucan, resulting in increased antioxidant-related gene expression plus enzymes and improved meat quality

Berberine inhibits SGIV replication by suppressing inflammatory response and oxidative stress

Singapore grouper iridovirus (SGIV) is one of the major infectious diseases responsible for high mortality and huge economic losses in the grouper aquaculture industry. Berberine (BBR), a naturally occurring plant alkaloid, is a phytochemical having a variety of biological properties, such as antiviral, antioxidant, and anti-inflammatory effects. In this work, we used an in vitro model based on Western blot, ROS fluorescence probe, and real-time quantitative PCR (qRT-PCR) to examine the antiviral qualities of BBR against SGIV. The outcomes demonstrated that varying BBR concentrations could significantly inhibit the replication of SGIV. In addition, BBR greatly inhibited the production of genes associated with pro-inflammatory cytokines in SGIV-infected or SGIV-uninfected GS cells based on qRT-PCR data. Subsequent investigations demonstrated that BBR suppressed the expression of the promoter activity of NF-κB and NF-κB-p65 protein. Additionally, BBR reduced the phosphorylation of ERK 1/2, JNK, and p38. Furthermore, BBR also inhibits SGIV-induced ROS production by upregulating the expression of antioxidant-related genes. In conclusion, BBR is a viable therapy option for SGIV infection due to its antiviral properties.

Effects of nuciferine on growth performance, body composition, lipid metabolism and antioxidant capacity of juvenile large yellow croaker (Larmichthys crocea) fed diets with high palm oil level

This study was conducted to explore the impacts of nuciferine (NF) on growth performance, body composition, lipid metabolism and antioxidant capacity of juvenile large yellow croaker (Larmichthys crocea) fed diets with high palm oil level. Five diets were formulated with fish oil (FO), palm oil (PO) and PO supplemented with 0.01%NF, 0.05%NF and 0.25%NF. After feeding for 10 weeks, the results showed that the growth performance (final body weight and specific growth rate) exhibited a significantly quadratic trend with the increase of NF addition, and the maximums were observed in PO supplemented with 0.05%NF group. Moreover, dietary PO diet with NF addition relieved the abnormal lipid accumulation (hepatosomatic index, crude lipid and triglyceride contents, total cholesterol contents, low-density lipoprotein-cholesterol contents in liver and serum) induced by PO diet. Simultaneously, NF addition inhibited PO diet-induced increase in the mRNA expressions of lipid synthesis-related genes and PO diet-induced activation of SREBP1. In addition, dietary PO diet with NF addition impeded PO diet-induced increase of hepatic MDA contents and recovered PO diet-induced inhibition of SOD and CAT activities. Similarly, NF addition alleviated PO diet-induced decrease in the mRNA expressions of antioxidant-related genes and PO diet-induced inhibition of SOD protein expression. Thus, addition of NF in PO diet might effectively ameliorate PO diet-induced growth retardation, lipid metabolism disorder and oxidative stress in croaker, and the optimal addition dose of NF in PO diet was approximately 0.05%.

Effects of antibacterial peptide microcin J25 on growth performance, antioxidant capacity, intestinal barrier function and intestinal microbiota in pigeon squabs

The present study aims to evaluate the effects of the antibacterial peptide Microcin J25 (ABP MccJ25) on growth performance, intestinal barrier function, antioxidant capacity and ileal microbiota in pigeon squabs. The 80 pairs of American Silver King pigeon parents and 240 healthy day 1 squabs were randomly divided into four groups, including 4 parent pairs and 12 pigeon squabs per group with 5 replicates. Parent pigeons were fed a basal diet (CON) or the addition of 100, 200, 300 mg/kg of ABP MccJ25 (ABP100, ABP200, and ABP300, respectively) for 4 weeks. Results showed that the group ABP200 remarkably reduced the feed conversion ratio and increased average daily gain in pigeon squabs from 7 to 21 d (p < 0.05), and improved serum antioxidant capacity in pigeon squabs from 14 to 28 d (p < 0.05). Villus height and the ratio of villus height to crypt depth in the duodenum, jejunum, and ileum were significantly increased in groups ABP200 and ABP300 (p < 0.05). In addition, group ABP200 obviously up-regulated the relative mRNA expression of intestinal antioxidant-related genes and barrier-related genes (p < 0.05). Moreover, the proportion of beneficial bacteria including Firmicutes and Lactobacillales, which were favourable to the intestinal health of pigeon squabs increased noticeably (p < 0.05). Taken together, these results indicated that group ABP200 enhanced antioxidant capacity, strengthened intestinal barrier function, and facilitated the regulation of intestinal microbiota, leading to the improvement in growth performance of pigeon squabs. Therefore, this study provides support for the application of 200 mg/kg ABP MccJ25 in pigeon production.

Study of the effect of calcium signal participating in the antioxidant mechanism of yeast under high-sugar environment.

Saccharomyces cerevisiae is susceptible to high-sugar stress in the production of bioethanol, wine and bread. Calcium signal is widely involved in various physiological and metabolic activities of cells. The present study aimed to explore the effects of Ca2+ signal on the antioxidant mechanism of yeast during high-sugar fermentation. Compared to yeast without available Ca2+, yeast in the high glucose with Ca2+ group had higher dry weight, higher ethanol output at 12 and 24 h and higher glycerol output at 24 and 36 h. During the whole growth process, the trehalose synthesis capacity of yeast in the high glucose with Ca2+ group was lower and intracellular reactive oxygen species content was higher compared to yeast without available Ca2+. Intracellular malondialdehyde content of yeast under high glucose with Ca2+ was significantly lower than yeast under high glucose without available Ca2+ except for 6 h. The superoxide dismutase and catalase activities of yeast and glutathione content were higher in the high glucose with Ca2+ group compared to yeast in high glucose without available Ca2+. The expression levels of SOD1, GSH1, GPX2 genes were higher for high glucose without available Ca2+ at 6 h, while yeast in the high glucose with Ca2+ group had a higher expression of antioxidant-related genes except SOD1 and CTT1 at 12 h. The expression levels of antioxidant-related genes of yeast for high glucose with Ca2+ were higher at 24 h, and those of genes except SOD1 of yeast in the high glucose with Ca2+ group were higher at 36 h. High-glucose stress limited the growth of yeast, while a moderate extracellular Ca2+ signal could improve the antioxidant capacity of yeast in a high-glucose environment by regulating protectant metabolism and enhancing the antioxidant enzyme activity and expression of antioxidant genes in a high-sugar environment. © 2024 Society of Chemical Industry.

Elderberry diet enhances motor performance and reduces neuroinflammation-induced cell death in cerebellar ataxia rat models

Cerebellar ataxia (CA) is a condition in which cerebellar dysfunction results in movement disorders such as dysmetria, synergy and dysdiadochokinesia. This study investigates the therapeutic effects of elderberry (EB) diet on the 3-acetylpyridine-induced (3-AP) CA rat model. First, CA rat models were generated by 3-AP administration followed by elderberry diet treatment containing 2 % EB for 8 consecutive weeks. Motor performance, electromyographic activity and gene expression were then evaluated. The number of Purkinje neurons were evaluated by stereological methods. Immunohistochemistry for the microgliosis, astrogliosis and apoptosis marker caspase-3 was also performed. In addition, the morphology of microglia and astrocytes was assessed using the Sholl analysis method. The results showed that EB diet administration in a 3-AP ataxia model improved motor coordination, locomotor activity and neuro-muscular function, prevented Purkinje neurons degeneration, increased microglia and astrocyte complexity and reduced cell soma size. Moreover, EB diet administration decreased apoptosis in cerebellum of 3-AP ataxic model. In addition, elderberry diet treatment decreased the expression of inflammatory, apoptotic and necroptotic genes and increased the expression of antioxidant-related genes. The results suggest that the EB diet attenuates 3-AP-induced neuroinflammation leading to cell death and improves motor performance. Thus, the EB diet could be used as a therapeutic procedure for CA due to its neuroprotective effects.

Effects of Dietary Aflatoxin B1 on Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) Growth, Intestinal Health, and Muscle Quality.

This study investigated the effects of varying doses of dietary aflatoxin B1 (AFB1) on the growth, intestinal health, and muscle quality of hybrid grouper. Four diets with varying AFB1 concentrations (0, 30, 445, and 2,230 μg kg-1) were used. Elevating AFB1 concentrations led to a decline in growth indexes, specifically the weight gain rate and the specific growth rate, although the survival rate remained unchanged. Morphological indicators showed a dose-dependent decline with AFB1 exposure. Intestinal MDA content and hindgut reactive oxygen species (ROS) levels increased, while antioxidant indexes and digestive enzymes decreased with higher AFB1 levels. AFB1 negatively influenced hindgut tight junction protein and antioxidant-related gene expression while promoting inflammation-related gene expression. The presence of AFB1 in the experiment led to a decrease in beneficial intestinal bacteria, such as Prevotella, and an increase in harmful intestinal bacteria, such as Prevotellaceae_NK3B31_group. Muscle lipid and unsaturated fatty acid content significantly decreased, while muscle protein and liver AFB1 content increased dramatically with higher AFB1 concentrations. AFB1 caused myofibrillar cleavage and myofilament damage, leading to increased spaces between muscle fibers. In conclusion, diets with AFB1 levels exceeding 30 μg kg-1 inhibited hybrid grouper growth, while levels surpassing 445 μg kg-1 resulted in hindgut ROS accumulation, inflammation, elevated intestinal permeability, reduced digestive enzyme activity, and compromised muscle quality.

The Antioxidant Salidroside Ameliorates the Quality of Postovulatory Aged Oocyte and Embryo Development in Mice.

Postovulatory aging is known to impair the oocyte quality and embryo development due to oxidative stress in many different animal models, which reduces the success rate or pregnancy rate in human assisted reproductive technology (ART) and livestock timed artificial insemination (TAI), respectively. Salidroside (SAL), a phenylpropanoid glycoside, has been shown to exert antioxidant and antitumor effects. This study aimed to investigate whether SAL supplementation could delay the postovulatory oocyte aging process by alleviating oxidative stress. Here, we show that SAL supplementation decreases the malformation rate and recovers mitochondrial dysfunction including mitochondrial distribution, mitochondrial membrane potential (ΔΨ) and ATP content in aged oocytes. In addition, SAL treatment alleviates postovulatory aging-caused oxidative stress such as higher reactive oxygen species (ROS) level, lower glutathione (GSH) content and a reduced expression of antioxidant-related genes. Moreover, the cytoplasmic calcium ([Ca2+]c) and mitochondrial calcium ([Ca2+]mt) of SAL-treated oocytes return to normal levels. Notably, SAL suppresses the aging-induced DNA damage, early apoptosis and improves spindle assembly in aged oocytes, ultimately elevating the embryo developmental rates and embryo quality. Finally, the RNA-seq and confirmatory experience showed that SAL promotes protective autophagy in aged oocytes by activating the MAPK pathway. Taken together, our research suggests that supplementing SAL is an effective and feasible method for preventing postovulatory aging and preserving the oocyte quality, which potentially contributes to improving the successful rate of ART or TAI.

MiR-15b-5p promotes HgCl2-induced chicken embryo kidney cells ferroptosis by targeting β-TrCP-mediated ATF4 ubiquitin degradation

Mercuric chloride (HgCl2), a widespread environmental pollutant, induces ferroptosis in chicken embryonic kidney (CEK) cells. Whereas activating transcription factor 4 (ATF4), a critical mediator of oxidative homeostasis, plays a dual role in ferroptosis, but its precise mechanisms in HgCl2-induced ferroptosis remain elusive. This study aims to investigate the function and molecular mechanism of ATF4 in HgCl2-induced ferroptosis. Our results revealed that ATF4 was downregulated during HgCl2-induced ferroptosis in CEK cells. Surprisingly, HgCl2 exposure has no significant impact on ATF4 mRNA level. Further investigation indicated that HgCl2 enhanced the expression of the E3 ligase beta-transducin repeat-containing protein (β-TrCP) and increased ATF4 ubiquitination. Subsequent findings identified that miR-15b-5p as an upstream modulator of β-TrCP, with miR-15b-5p downregulation observed in HgCl2-exposed CEK cells. Importantly, miR-15b-5p mimics suppressed β-TrCP expression and reversed HgCl2-induced cellular ferroptosis. Mechanistically, HgCl2 inhibited miR-15b-5p, and promoted β-TrCP-mediated ubiquitin degradation of ATF4, thereby inhibited the expression of antioxidant-related target genes and promoted ferroptosis. In conclusion, our study highlighted the crucial role of the miR-15b-5p/β-TrCP/ATF4 axis in HgCl2-induced nephrotoxicity, offering a new therapeutic target for understanding the mechanism of HgCl2 nephrotoxicity.

Pre-hatch thermal manipulation of embryos and post-hatch baicalein supplementation mitigated heat stress in broiler chickens

BackgroundHigh environmental temperatures induce heat stress in broiler chickens, affecting their health and production performance. Several dietary, managerial, and genetics strategies have been tested with some success in mitigating heat stress (HS) in broilers. Developing novel HS mitigation strategies for sustaining broiler production is critically needed. This study investigated the effects of pre-hatch thermal manipulation (TM) and post-hatch baicalein supplementation on growth performance and health parameters in heat-stressed broilers.ResultsSix hundred fertile Cobb 500 eggs were incubated for 21 d. After candling on embryonic day (ED) 10, 238 eggs were thermally manipulated at 38.5 °C with 55% relative humidity (RH) from ED 12 to 18, then transferred to the hatcher (ED 19 to 21, standard temperature) and 236 eggs were incubated at a controlled temperature (37.5 °C) till hatch. After hatch, 180-day-old chicks from both groups were raised in 36 pens (n = 10 birds/pen, 6 replicates per treatment). The treatments were: 1) Control, 2) TM, 3) control heat stress (CHS), 4) thermal manipulation heat stress (TMHS), 5) control heat stress supplement (CHSS), and 6) thermal manipulation heat stress supplement (TMHSS). All birds were raised under the standard environment for 21 d, followed by chronic heat stress from d 22 to 35 (32–33 °C for 8 h) in the CHS, TMHS, CHSS, and TMHSS groups. A thermoneutral (22–24 °C) environment was maintained in the Control and TM groups. RH was constant (50% ± 5%) throughout the trial. All the data were analyzed using one-way ANOVA in R and GraphPad software at P < 0.05 and are presented as mean ± SEM. Heat stress significantly decreased (P < 0.05) the final body weight and ADG in CHS and TMHS groups compared to the other groups. Embryonic TM significantly increased (P < 0.05) the expression of heat shock protein-related genes (HSP70, HSP90, and HSPH1) and antioxidant-related genes (GPX1 and TXN). TMHS birds showed a significant increment (P < 0.05) in total cecal volatile fatty acid (VFA) concentration compared to the CHS birds. The cecal microbial analysis showed significant enrichment (P < 0.05) in alpha and beta diversity and Coprococcus in the TMHSS group.ConclusionsPre-hatch TM and post-hatch baicalein supplementation in heat-stressed birds mitigate the detrimental effects of heat stress on chickens' growth performance, upregulate favorable gene expression, increase VFA production, and promote gut health by increasing beneficial microbial communities.

Combined exposure of nano-titanium dioxide and polystyrene nanoplastics exacerbate oxidative stress-induced liver injury in mice by regulating the Keap-1/Nrf2/ARE pathway.

It is well known that polystyrene nanoplastics (PS-NaP) and nano-titanium dioxide (TiO2 NPs) are frequently co-appeared in daily life and can cause liver injury when they accumulate in the liver. Nonetheless, the combined toxicological impacts and potential molecular mechanisms of PS-NaP and TiO2 NPs in the hepatic system have not been revealed. Thus, we conducted experiments on C57BL/6 mice exposed to PS-NaP or/and TiO2 NPs for 4 weeks. The findings suggested that PS-NaP and TiO2 NPs co-exposed significantly altered the hepatic function parameters, levels of antioxidant-related enzymes and genes expression of Keap-1/Nrf2/ARE signaling pathway, as well as significantly increased the hepatic Ti contents, aggravated hepatic pathological and oxidative stress (OS) damage compared with individual exposure to PS-NaP or TiO2 NPs. Using N-Acetyl-L-cysteine (NAC), an OS inhibitor, we further demonstrated that OS played a pivotal role in coexposure-induced liver injury. NAC reduced the levels of OS in mice, which mitigated co-exposure-induced liver injury. Taken together, we proposed that PS-NaP and TiO2 NPs co-exposed activated the Keap-1, then inhibited the recognition of Nrf2 and ARE, consequently exacerbated liver injury. These findings shed light on the co-toxicity and potential mechanism of nanoplastics and nanoparticles, which informed the risk assessment of human exposure to environmental pollutants.