Glutathione Levels Research Articles

Antioxidative, anti-androgenic, and inhibitory activities of ethanolic extract of Annona muricata leaf on sex hormones-induced benign prostate hyperplasia through in vivo and in silico studies.

Benign prostate hyperplasia (BPH) remains one of the major age-related urological problems in the world. Annona muricata (soursop) leaf has been reported to exhibit antiproliferative, antioxidant, and anti-inflammatory activities, among others, in the literature. Here, we aimed to unravel the antioxidative, antiandrogenic, and inhibitory activities of the ethanol extract of Annona muricata leaf (EEAML) on sex hormones-induced benign prostate hyperplasia (BPH) through in vivo and in silico studies. Thirty-six male rats were segmented into six groups of six animals each, the control group received water, and the BPH group and the remaining four groups were parentally infused with testosterone (T) and oestradiol (E2) (0.08 and 0.04 mg/Kgbwt) once daily for 28 days to induce BPH. After that, the control and BPH groups received water and normal saline, while the remaining four groups received finasteride (FIN) (0.1 mg/kgbwt) and EEAML (200, 400, and 800 mg/kgbwt) for another 28 days before sacrifice, and serum was collected for biochemical analysis. Additionally, the active ingredients of EEAML were identified using a Gas Chromatography Flame Ionisation Detector (GC-FID) followed by molecular docking (MD) against the human androgen receptor (hAR) target, and ADMET analysis of selected EEAML compounds was carried out. EEAML (200, 400, and 800 mg/kgbwt) restored the T and E2-induced depletion of reduced glutathione level, superoxide dismutase and catalase activities, and elevation of malondialdehyde, prostate-specific antigen, testosterone, and dihydrotestosterone levels in the serum of BPH rats. GC-FID analysis of EEAML showed the presence of 21 compounds from which 15 compounds were subjected to MD revealing that flavone, followed by ribalinidine, flavonone, anthocyanin, and naringenin displayed desirable binding affinities against the hAR target. ADMET analysis of these top-five EEAML compounds revealed that they were excellent oral bioavailable drug candidates with predicted minimal toxicities. In conclusion, EEAML exhibited antioxidative, antiandrogenic, and inhibitory activities owing to its phytoconstituents, which in turn could serve as drug templates for much better efficacy.

Isoorientin Promotes Early Porcine Embryonic Development by Alleviating Oxidative Stress and Improving Lipid Metabolism

Isoorientin (ISO) is a natural lignan glycoside flavonoid found in various plants, including Charcot and Stonecrop. ISO exhibits diverse physiological and pharmacological effects, such as antioxidative, anti-inflammatory, hepatoprotective, antiviral, antianxiety, and anti-myocardial ischaemic properties, as well as lipid metabolism regulation. This study investigated the impact of ISO supplementation on oxidative stress and lipid accumulation in porcine early embryos, along with its underlying mechanisms. Porcine embryos were cultured in vitro under different concentrations of ISO (0, 1, 10, and 100 nM). The results revealed that 10 nM ISO significantly enhanced the blastocyst rate and total embryonic cell count in vitro. ISO-treated embryos exhibited reduced reactive oxygen species levels and elevated glutathione levels compared to the untreated group. In addition, ISO treatment significantly increased the expression of the key antioxidant regulator Nrf2, improved mitochondrial function, and reduced lipid droplet accumulation. Concurrently, early embryo autophagy and apoptosis levels decreased. Furthermore, ISO treatment upregulated antioxidant-related genes (SOD1, SOD2, and CAT) and mitochondrial biogenesis related genes (NRF1, NRF2, and SIRT1), while downregulating lipid synthesis-related genes (SREBP1 and FASN). Additionally, lipid hydrolysis-related genes (ACADS) were elevated. These findings collectively suggest that ISO may facilitate early embryonic development in pigs by ameliorating oxidative stress and lipid metabolism.

Cardioprotective Effects of Ursodeoxycholic Acid in Isoprenaline-Induced Myocardial Injury in Rats.

Patients suffering from cholelithiasis have an increased risk of developing cardiovascular complications, particularly ischemic myocardial disease. Ursodeoxycholic acid (UDCA), already used in clinical practice for the treatment of cholelithiasis and related conditions, has proven antioxidative, anti-inflammatory, and cytoprotective effects. Therefore, the aim of this study was to investigate the cardioprotective effect of UDCA pre-treatment on isoprenaline-induced myocardial injury in rats. Male Wistar albino rats were randomized into four groups. Animals were pre-treated for 10 days with propylene glycol + saline on days 9 and 10 (control), 10 days with propylene glycol + isoprenaline on days 9 and 10 (I group), 10 days with UDCA + saline on days 9 and 10 (UDCA group), and 10 days with UDCA + isoprenaline on days 9 and 10 (UDCA + I group). UDCA pre-treatment significantly reduced values of high-sensitivity troponin I (hsTnI) and aspartate aminotransferase (AST) cardiac markers (p < 0.001 and p < 0.01, respectively). The value of thiobarbituric acid reactive substances (TBARS) was also decreased in the UDCA + I group compared to the I group (p < 0.001). UDCA also significantly increased glutathione (GSH) levels, while showing a tendency to increase levels of superoxide dismutase (SOD) and catalase (CAT). The level of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) expression, a key regulatory gene of inflammation, was diminished when UDCA was administered. A reduction of cardiac damage was also observed in the UDCA pre-treated group. In conclusion, UDCA pre-treatment showed a cardioprotective effect on isoprenaline-induced myocardial injury in rats, primarily by reducing oxidative stress and inflammation.

Disulfiram-Copper Potentiates Anticancer Efficacy of Standard Chemo-therapy Drugs in Bladder Cancer Animal Model through ROS-Autophagy-Ferroptosis Signalling Cascade.

Cost-effective management of Urinary Bladder Cancer (UBC) is an unmet need. Our study aims to demonstrate the efficacy of a drug repurposing strategy by using disulfiram (DSF) and copper gluconate (Cu) as an add-on treatment combination to traditional GC-based chemother-apy against N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced UBC mice (C57J) model. Male C57BL/6J mice were given 0.05% BBN in drinking water ad libitum, and tumour for-mation was verified by histological and physical evaluation. Animals were subsequently divided into eight groups and received treatment with different drug combinations. Control animals received only ve-hicle (DMSO). At the end of the treatment schedule, the bladder tumour was excised and further used to check the expression (mRNA and protein) of ALDH1 isoenzymes using qRT-PCR, western blot, and IHC methods. Autophagy induction was assessed by quantifying the expression of LC3B and SQSTM1/p62 proteins through IHC. Biochemical analysis of superoxide dismutase (SOD), reduced glutathione (GSH), and lipid peroxidation levels in the freshly isolated tumours was performed to check the alterations in the antioxidant system caused by combination treatment. We observed significant induction of an invasive form of bladder cancer in the mice after nine-teen weeks of BBN exposure. The animals began exhibiting early indications of inflammatory alterations as early as the sixth week following BBN treatment. Furthermore, the wet bladder weight and overall tu-mour burden were significantly decreased (p&#60; 0.0001) by DSF-Cu co-treatment in addition to the GC-based chemotherapy. Real-time PCR analysis revealed that treatment with disulfiram and copper glu-conate significantly decreased (p&#60;0.0001) the mRNA expression of ALDH1 isoenzymes. Comparing the triple drug combination group (GC+DSF-Cu) to the untreated mice, a significant rise in LC3B puncta (p&#60;0.0001) and a decrease in P62/SQSTM1 (p=0.0002) were noted, indicating the induction of autophagy flux in the add-on group. When GC+DSF-Cu treated mice were compared to the untreated tumour group, a substantial decrease in ALDH1/2 protein expression was observed (p= 0.0029 in IHC and p&#60;0.0001 in western blot). Lipid peroxidation was significantly higher (p&#60;0.0001) in the triple drug combination group than in untreated mice. There was a simultaneous decrease in reduced glutathione (GSH) and en-zyme superoxide dismutase (SOD) levels (p&#60;0.0001), which strongly suggests the generation of reactive oxygen species and induction of ferroptotic cell death in the add-on therapy group. Additionally, in both IHC and western blot assays, ALDH1A3 expression was found to be significantly increased (p=0.0033, &#60;0.0001 respectively) in GC+DSF-Cu treated mice relative to the untreated group, suggesting a potential connection between the ferroptosis pathway and ALDH1A3 overexpression. It was found that disulfiram with copper treatment inhibits bladder tumour growth through ferroptosis-mediated ROS induction, which further activates the process of autophagy. Our results prove that DSF-Cu can be an effective add-on therapy along with the standard chemotherapy drugs for the treatment of UBC.

Pseudomonas aeruginosa LasR-deficient mutants have increased methylglyoxal and hydrogen peroxide sensitivity due to low intracellular glutathione.

Methylglyoxal is a highly reactive metabolite that is detected in various disease states, including those where Pseudomonas aeruginosa is present and MG resistance requires the glutathione-dependent glyoxalase enzyme GloA3 enzyme. This study reveals that P . aeruginosa strains with LasR mutations, which are commonly found in clinical isolates, are more sensitive to methylglyoxal (MG) and hydrogen peroxide due to lower intracellular glutathione levels and high activity of the CbrAB-Crc regulatory pathway. This could be significant for understanding the selective pressures that drive P. aeruginosa evolution in infection sites, as well as a better understanding of LasR-strain metabolism in infections such as those associated with cystic fibrosis.

Evaluation of the Impact of Pistacia atlantica Desf. Oleo-gum Resin on Complete Freund’s Adjuvant-induced Arthritis in a Wistar Rat Model

Background: Pistacia atlantica Desf., also known as Atlantic pistachio or Mt. Atlas mastic tree, has been used in traditional medicine for its anti-inflammatory properties, helping to reduce inflammation in conditions like arthritis. Additionally, the tree's resin possesses antioxidant properties that can help protect against oxidative stress. The anti-arthritic efficacy of Pistacia atlantica oleo-gum resin extract (PAG) remains unvalidated, as its potential therapeutic benefits have not been substantiated. In the current investigation, we have substantiated the inhibitory properties of the PAG in mitigating the development of rheumatoid arthritis induced by Complete Freund's Adjuvant in rats. Methods: Male Wistar rats weighing 160±10 g were subjected to immunization through intradermal injection of 0.1 mL of Complete Freund's Adjuvant (CFA) into the left hind metatarsal footpad. Following the induction of adjuvant arthritis, oral administration of PAG was initiated at doses of 10, 400, and 800 mg/kg once a day. Prednisolone (10 mg/kg, daily) was employed as a positive control in this study. Various parameters, including paw swelling, arthritic index, body weight loss, spleen index, thymus index, serum cytokines, inflammatory mediators, and histological alterations, as well as nitrite glutathione and catalase level, were measured. Furthermore, the expression and activity of matrix metalloproteinase (MMP)-2 and MMP-9 were evaluated. Results: PAG exhibited noteworthy efficacy in attenuating paw swelling and reducing the arthritic index. Moreover, it resulted in enhanced body weight loss and a consequential reduction in the thymus index. PAG (400mg/kg) significantly decreased the level of serum nitrite in the intervention group, as well as the levels of serum glutathione. On the other hand, catalase activity increased, the activity of MMP-9 decreased, and the level of MMP-2 increased compared to the intervention group. The histopathological results revealed that PAG (at doses of 400 mg/kg) had the best therapeutic effects. Conclusion: In conclusion, the present study confirms the anti-arthritic activity of PAG in rats. The extract of PAG was found to prevent rheumatoid arthritis induced by Complete Freund's Adjuvant, as evidenced by the significant suppression of paw swelling and arthritic index.

Making bloodwork work: the impact of sample collection, processing, and storage on plasma glutathione measurement, and implications for translation

Psychiatry has traditionally focused on the study of neurons and neurotransmitter physiology in the pathophysiology and treatment of psychiatric disorders. A growing literature highlights REDOX imbalance (a state in which demand for antioxidants surpasses their bioavailability) as a common pathophysiology for a diverse array of brain conditions (e.g., trichotillomania, schizophrenia, autism, Parkinson’s disease). REDOX imbalance is typically measured via plasma glutathione, as glutathione is critical to the adaptive antioxidant response in the brain. Accordingly, glutathione, its precursors, and/or metabolites serve as biomarkers of disease risk, therapeutic targets, and measures of treatment response. However, as with any emerging field, there are currently several different methods for collection, processing, storage, and calculation of summary measures of plasma glutathione metabolism, within and between preclinical and clinical research. The lack of evidence-based best-practice methodology hampers reproducibility (preclinical or clinical), and translation (between preclinical and clinical work). To address this methodological need, here we used a repeated measures within-subject design to investigate how sample preparation (type of anticoagulant used during blood collection, deproteinization status, and storage temperature) affects plasma glutathione levels. Accordingly, we collected whole blood from mice (N = 13), and then, using a commercially available kit, quantified glutathione in plasma stored in four different ways. Presuming that these preparation conditions and post-processing calculations are unimportant, we would expect to see no difference in glutathione levels and summary measures from the same sample. However, we found each of these variables to significantly alter quantified glutathione levels. Accordingly, we propose a vital, gold-standard methodology for both sample collection, processing, and storage of plasma used for glutathione quantification and for summary calculations of glutathione that can be used preclinically and clinically, thus yielding more streamlined translation.

α-Arbutin ameliorates UVA-induced photoaging through regulation of the SIRT3/PGC-1α pathway.

Owing to its tyrosinase inhibitory activity, α-arbutin has been added to several skin care products as a skin-lightening agent. However, the protective effect of α-arbutin against ultraviolet A (UVA)-induced photoaging has not been well investigated. The present study was designed to investigate the photoprotective effect and mechanism of α-arbutin against UVA-induced photoaging. In vitro experiments, HaCaT cells were treated with UVA at a dose of 3J/cm2 to evaluate the anti-photoaging effect of α-arbutin. α-Arbutin was found to exhibit a strong antioxidant effect by increasing glutathione (GSH) level and inhibiting reactive oxygen species (ROS) production. Meanwhile, α-arbutin markedly improved the expression of sirtuin 3 (SIRT3) and peroxisome proliferator-activated receptor γ coactivator 1 α (PGC-1α) proteins, initiating downstream signaling to increase mitochondrial membrane potential and mediate mitochondrial biogenesis, and improve mitochondrial structure significantly. In vivo analysis, the mice with shaved back hair were irradiated with a cumulative UVA dose of 10J/cm2 and a cumulative ultraviolet B (UVB) dose of 0.63J/cm2. The animal experiments demonstrated that α-arbutin increased the expression of SIRT3 and PGC-1α proteins in the back skin of mice, thereby reducing UV-induced skin damage. In conclusion, α-arbutin protects HaCaT cells and mice from UVA damage by regulating SIRT3/PGC-1α signaling pathway.

Tire Rubber Based Microplastic Particles Cause Adverse on Quality Parameters of Rainbow Trout Sperm Cells.

In the present study, we aimed to determine the parameters of oxidative stress markers, motility and kinematics of rainbow trout (Oncorhynchus mykiss) sperm cells exposed to different doses (0.001, 0.01, 0.1, 1.0, and 10mg/L, in vitro 4h) of tire rubber based microplastic particles (TRMP-Ps) the leachates procedure of rubber pieces. First of all, TRMP-Ps were prepared by abrasion method in accordance with the literature. Structural and morphological features of TRMP-Ps were determined by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) methods, respectively. Energy dispersive X-ray (EDX) analysis technique was used to characterize the elemental composition of TRMP-Ps. Particle size of microplastic structures was measured hydrodynamically with dynamic light scattering analysis (DLS). After exposure, the effect of TRMP-Ps was defined by the observations of kinematics and antioxidant activities in sperm cells. Our findings showed that the straight line velocity, the curvilinear velocity, the angular path velocity, and the amplitude of lateral displacement of sperm cells decreased. Moreover, while the level of superoxide dismutase decreased dose-dependently against the toxicity of TRMP-Ps, no significant change was observed in the levels of malondialdehyde and total glutathione. The 4-h median effective concentrations (EC50) of TRMP-Ps based on mobility parameters of sperm ranged from 0.31mg/L for reduced straight line velocity of sperm cells to 0.51mg/L for reduced amplitude of lateral displacement of the spermatozoa head. Therefore, we concluded that TRMP-Ps can be a risk for the reproduction cycle of fish in aquatic environments.

Activation of GABABR Attenuates Intestinal Inflammation by Reducing Oxidative Stress through Modulating the TLR4/MyD88/NLRP3 Pathway and Gut Microbiota Abundance.

Oxidative stress emerges as a prominent factor in the onset and progression of intestinal inflammation, primarily due to its critical role in damaging cells and tissues. GABAergic signaling is important in the occurrence and development of various intestinal disorders, yet its effect on oxidative stress remains unclear. We attempted to assess whether GABAergic signaling participated in the regulation of oxidative stress during enteritis. The results showed that lipopolysaccharide (LPS) significantly decreased γ-aminobutyric acid (GABA) levels in the ileal tissues of mice. Interestingly, the application of GABA significantly repressed the shedding of intestinal mucosal epithelial cells and inflammatory cell infiltration, inhibited the expressions of proinflammatory factors, including granulocyte colony-stimulating factor and granulocyte-macrophage colony stimulating factor, and enhanced the levels of anti-inflammatory cytokines interleukin (IL)-4 and IL-10, indicating that GABA could alleviate enteritis in mice. This observation was further supported by transcriptome sequencing, revealing a total of 271 differentially expressed genes, which exhibited a marked enrichment of inflammatory and immune-related pathways, alongside a prominent enhancement of GABA B receptor (GABABR) signaling following GABA administration. Effectively, Baclofen pretreatment alleviated intestinal mucosal damage in LPS-induced mice, suppressed proinflammatory cytokines IL-1β, IL-6, and tumor necrosis factor alpha expressions, and boosted total antioxidant capacity, superoxide dismutase (SOD), and glutathione (GSH) levels. Moreover, Baclofen notably enhanced the viability of LPS-stimulated IPEC-J2 cells, contracted the proinflammatory secretion factors, and reinforced SOD, GSH, and catalase levels, emphasizing the anti-inflammatory and antioxidant effects associated with GABABR activation. Mechanistically, Baclofen restrained the mRNA and protein levels of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3), and inducible nitric oxide synthase, while elevating nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 in both mice and IPEC-J2 cells, indicating that activating GABABR strengthened antioxidant abilities by interrupting the TLR4/MyD88/NLRP3 pathway. Furthermore, 16S rDNA analysis demonstrated that Baclofen increased the relative abundance of probiotic, particularly Lactobacillus, renowned for its antioxidant properties, while reducing the relative richness of harmful bacteria, predominantly Enterobacteriaceae, suggesting that GABABR signaling may have contributed to reversing intestinal flora imbalances to relieve oxidative stress in LPS-induced mice. Our study identified previously unappreciated roles for GABABR signaling in constricting oxidative stress to attenuate enteritis, thus offering novel insights for the treatment of intestinal inflammation.

Potential Protective Effect of Hesperidin (Vitamin P) against Glyphosate-Induced Spermatogenesis Damage in Male Rats: Biochemical and Histopathological Findings on Reproductive Parameters.

The aim of this study was to investigate the effectiveness of hesperidin (HES) on testicular histopathological changes, biochemical changes, and semen characteristics in rats exposed to glyphosate (GLP). The control group was given a normal diet devoid of GLP and HES, the HES group was given 100 mg/kg/day HES with the normal diet, the GLP group was given GLP at the LD50/10 dose of normal feed, which was 787.85 mg/kg/day, and the GLP + HES group was given normal feed containing 787.85 mg/kg/day LD50/10 dose of GLP in addition to 100 mg/kg/day HES. GLP administration reduced sperm motility, sperm plasma membrane integrity, glutathione levels, and total antioxidant levels in the testicular tissues of rats. Moreover, it caused an increase in right testis and left epididymis weights, abnormal sperm counts, malondialdehyde levels, total oxidant status, and DNA damage. The HES treatment showed curative effects on these parameters. Furthermore, HES was effective in lessening the histopathological damage that was caused by GLP. The results showedthat HES protects spermatological parameters and DNA integrity, improves antioxidant defenses, and lowers the damage and lipid peroxidation caused by GLP in testicular tissue.

Studies on the role of moderate doses of ionizing radiation-induced cellular senescence in mouse lung tissue

Purpose To investigate the role of moderate doses of ionizing radiation-induced cellular senescence in mouse lung tissue and whole-body inflammation levels. Material and methods Forty-two C57BL/6J mice were randomly divided into the control group, the 1, 3, and 7 days after 2 Gy irradiation group, and the 1, 3, and 7 days after 4 Gy irradiation group, with six mice in each group. The histopathology, cellular senescence, oxidative-antioxidant, DNA damage repair, and inflammation-related indicators of irradiated mice were examined. Results Compared with the control group, the histopathological scores, the positive area of senescence-associated-β-galactosidase (SA-β-Gal) staining, and the mRNA levels of senescence-related genes in the lung tissues in all dose groups increased on 1, 3, and 7 days after irradiation. In peripheral blood, erythrocytes, leukocytes, platelets, hemoglobin, 8-hydroxydeoxyguanosine (8-OHdG), C-reactive protein, and other indicators showed a different trend in all dose groups. The levels of malondialdehyde(MDA), superoxide dismutase (SOD), glutathione (GSH), and 8-OHdG in the lung tissue showed different trends after 2 Gy and 4 Gy irradiation. The 8-Oxoguanine DNA glycosylase 1 (hOGG1) and O-6-methylguanine-DNA methyltransferase (MGMT) mRNA levels showed a trend of increasing and then decreasing. The levels of whole-body inflammation were significantly correlated with the levels of indicators related to cellular senescence and damage repair in the lung tissue of mice. Conclusions The moderate doses of ionizing radiation induce oxidative stress, and DNA damage and increase DNA repair gene expression in mouse lung tissue. The lung tissue cellular senescence correlates with the level of whole-body inflammation.

Effect of chlorine dioxide (ClO2) combined with different oxygen permeability package treatments on postharvest antioxidant activity and storage life of ‘Liaofeng’ grapes

To enhance the storage quality of ‘Liaofeng’ grapes, this research employed chlorine dioxide in collaboration with different package of preservative bags to explore its preservative effect on ‘Liaofeng’ grapes. After fumigating the grapes with 300 mg L−1 ClO2 solution, the control group was packaged in unpacked and PE bags, while the treatment group was packaged in three kinds of bags with different oxygen permeability, and stored in 4 °C and relative humidity controlled at 90%–95% for 90 days. The results showed that ClO2-GB package treatment significantly delayed the decline of grape hardness (P < 0.05), and decreased the weight loss, threshing, decay and peduncle browning index after storage for 90 days. ClO2-GB package treatment not only diminishes the levels of soluble solids content, titrable acidity, ascorbic acid and glutathione but also effectively suppresses the respiration rate in fruits. The contents of total phenols, flavonoids, anthocyanins and reduced glutathione after ClO2-GB treatment were significantly higher than those in unpackaged group (P < 0.05), and curbed the increase in relative electrical conductivity and malondialdehyde concentration. In comparison to ClO2-YTB and ClO2-YB treatments, ClO2-GB treatment demonstrated superior efficacy in modulating the antioxidant metabolism of grapes. It notably elevated the enzymatic activities of catalase, superoxide dismutase, ascorbate peroxidase, and glutathione reductase, while inhibiting the activities of peroxidase and polyphenol oxidase. The quality assessment of ‘Liaofeng’ grapes after 90 d storage period, conducted through principal component analysis (PCA), yielded the following ranking: ClO2-GB > ClO2-YB > ClO2-YTB > ClO2-PE > ClO2. This suggests that 300 mg L−1 ClO2 fumigation combined with different package treatments was more beneficial to the storage of ‘Liaofeng’ grape. ClO2-GB package treatment emerged as the most advantageous.

Bovine lactoferrin alleviates aflatoxin B1 induced hepatic and renal injury in broilers by mediating Nrf2 signaling pathway

Aflatoxin B1 (AFB1) a mycotoxin found in chicken feed that possess a global hazard to poultry health. However different potent compounds like bovine lactoferrin (bLF) may prove to be protective effects against AFB1. This study aims to explore the protective effect of bLF against AFB1-induced injury in the liver and kidney in broiler. For this purpose, 600 broilers chicks were randomly alienated into five groups (n=120 each): negative control; positive control (3mg/kg AFB1), and bLF high, medium, and low dosage groups (600 mg/kg, 300 mg/kg, and 150 mg/kg, respectively). The results highlight that AFB1 toxicity in birds exhibited low feed intake, reduction in weight gain, and a decrease in FCR while, bLF regulated these adverse effects. Meanwhile, AFB1 group showed higher levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) and lower levels of superoxide dismutase (SOD) and glutathione (GSHpx) in liver, while urea and creatinine were decline in kidney. Supplementation with bLF effectively controlled these biomarkers and control the negative effects of toxicity. Furthermore, hematoxylin and eosin (H&E) staining exhibited normal morphological structures within liver and kidney in the bLF treated groups, while degenerative changes were observed in AFB1 group. Similarly, bLF, decreased oxidative stress and thus prevented apoptosis in the liver and kidney cells of the birds. Whereas, mRNA level of mitochondrial apoptosis related gene including Bcl-2 (Bak and Bax), caspase-3 and caspase-9 was upregulated, while bcl2 gene were downregulated in AFB1 group. Dietary supplementation of bLF effectively normalizes the expression of these genes. AFB1 exposed birds shown to decrease gene expression level of the crucial component of Nrf2 pathway, responsible to regulate antioxidant defense. Interestingly, bLF reverse these detrimental effects of and restore the normal expression levels of Nrf2 pathway. Conclusively, our findings demonstrate that bLF mitigates the detrimental effects of AFB1, besides regulation of the apoptosis-related genes via mitochondrial pathways. These findings validate that the bLF (600 mg/kg) could be used as protective agent against AFB1-induced liver and kidney damage.

An antireductant approach ameliorates misfolded proinsulin-induced hyperglycemia and glucose intolerance in male Akita mice.

Protein folding in the endoplasmic reticulum (ER) requires a high ratio of oxidized to reduced glutathione (GSSG/rGSH). Since the GSSG/rGSH depends on total glutathione (tGSH = GSSG + rGSH) levels, we hypothesized that limiting GSH biosynthesis will ameliorate protein misfolding by enhancing the ER oxidative milieu. As a proof-of-concept, we used DL-buthionine-(S,R)-sulfoximine (BSO) to inhibit GSH biosynthesis in Akita mice, which are prone to proinsulin misfolding. We conducted a 2-week intervention to investigate if BSO was safe and a 6-week intervention to find its effect on glucose intolerance. In both cohorts, male heterozygous Akita (AK) and wild-type (WT) mice were continuously administered 15mM BSO. No adverse effects were observed on body weight, food intake, and water intake in either cohort. Unaltered levels of plasma aspartate and alanine aminotransferases, and cystatin-C, indicate that BSO was safe. BSO-induced decreases in tGSH were tissue-dependent with maximal effects in the kidneys, where it altered the expression of genes associated with GSH biosynthesis, redox status, and proteostasis. BSO treatment decreased random blood glucose levels to 80% and 67%of levels in untreated mice in short-term and long-term cohorts, respectively, and 6-h fasting blood glucose to 82% and 74%of levels in untreated mice, respectively. BSO also improved glucose tolerance by 37% in AK mice in the long-term cohort, without affecting insulin tolerance. Neither glucose tolerance nor insulin tolerance were affected in WT. Data indicate that BSO might treat misfolded proinsulin-induced glucose intolerance. Future studies should investigate the effect of BSO on proinsulin misfolding and if it improves glucose intolerance in individuals with Mutant Insulin Diabetes of Youth.

Intermittent fasting alerts neurotransmitters and oxidant/antioxidant status in the brain of rats

Several recent studies have attempted to understand how fasting has benefits for body health, especially the nervous system. To evaluate the impact of intermittent fasting on body weight, brain neurotransmitters, brain oxidative stress, and brain-derived neurotrophic factor (BDNF) in several areas of the brain, this study was conducted in rats. Thirty male Wistar rats were randomly divided into two groups. Group 1 (15 rats) served as the control and group 2 (15 rats) underwent intermittent fasting (IF; 24 h) for 1, 7, or 15 days. The findings demonstrated that intermittent fasting significantly reduced body weight. In this sense, brain monoamines and amino acids, namely dopamine, glutamate, aspartate, and oxidative stress markers (malondialdehyde and nitric oxide), decreased significantly after 1 day of IF. However, norepinephrine, serotonin, gamma-amino butyric acid, and glycine increased significantly. Additionally, glutathione levels were markedly elevated in IF. Surprisingly, the neuromodulatory effect of intermittent fasting fluctuates depending on the IF period. To support this fluctuation, BDNF levels increased after 1 day in the hippocampus and decreased after 15 days of intermittent fasting in all areas of the brain tested. In conclusion, our results show that intermittent fasting has beneficial influences on the brain; however, prolonged intermittent fasting can also induce some unfavorable physiological outcomes that prevent optimal neurological function.

Etomidate protects retinal ganglion cells from hydrogen peroxide-induced injury via Nrf2/HO-1 pathway.

To determine whether etomidate (ET) has a protective effect on retinal ganglion cells (RGCs) injured with hydrogen peroxide (H2O2) and to explore the potential mechanism underlying the antioxidative stress effect of ET. Cultured RGCs were identified by double immunofluorescent labeling of microtubule-associated protein 2 and Thy1.1. An injury model of H2O2-induced RGCs oxidative stress was established in vitro. Cells were pretreated with different concentrations of ET (1, 5, and 10 µmol/L) for 4h, followed by further exposure to H2O2 at 1000 µmol/L. Cell counting kit 8 and Annexin V/propidium iodide assays were applied to detect the viabilities and apoptosis rates of the RGCs at 12, 24, and 48h after H2O2 stimulation. The levels of nitric oxide, malondialdehyde, and glutathione in culture media were measured at these time points. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to observe the effects of ET on the messenger RNA and protein expression of inducible nitric oxide synthase (iNOS), nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), glutathione peroxidase 1 and the level of conjugated acrolein in RGCs at 12, 24, and 48h after H2O2 stimulation and in the retina at 12h after optic nerve transection (ONT). The applications of 5 and 10 µmol/L of ET significantly increased the viability of RGCs. Results from qRT-PCR indicated a decrease in the expression of iNOS and an increase in the expressions of Nrf2 and HO-1 in ET-pretreated RGCs at 12, 24 and 48h after H2O2 stimulation, as well as in ET-treated retinas at 12h after ONT. Western blot analysis revealed a decrease in the expression of iNOS and levels of conjugated acrolein, along with an increase in the expressions of Nrf2 and HO-1 in ET-pretreated RGCs in vitro and ET-treated retinas in vivo. ET is a neuroprotective agent in primary cultured RGCs injured by H2O2. The effect of ET is dose-dependent with the greatest effect being at 10 µmol/L. ET plays an antioxidant role by inhibiting iNOS, up-regulating Nrf2/HO-1, decreasing the production of acrolein, and increasing the scavenge of acrolein.

Neuroprotective Effect of Codonopsis pilosula Polysaccharide on Aβ25-35-Induced Damage in PC12 Cells via the p38MAPK Signaling Pathways.

Plant polysaccharides have attracted increasing attention due to their high efficiency and low toxicity. Codonopsis pilosula polysaccharide (CPP) is an essential substance extracted from Codonopsis pilosula, known for its excellent antioxidant and neuroprotective effects. However, it is still unclear how CPP improves nerve protection and what its underlying molecular mechanisms are. This study aimed to investigate the neuroprotective effect of CPP on Aβ25-35-induced damage in PC12 cells and its underlying molecular mechanisms. The neuroprotective effect of CPP was evaluated using Aβ25-35-induced damage in pheochFfromocytoma (PC12) cells as an in vitro cell model. The cells were treated with CPP alone or in combination with SB203580 (an inhibitor of p38MAPK) in Aβ25-35 culture. The cell viability was assessed using a 3-(4,5-Dimethylthiazol-2-yl)-2,diphenyltetrazolium (MTT) assay. Furthermore, reactive oxygen species (ROS) were detected using flow cytometry. The production levels of intracellular superoxide dismutase (SOD), dismutase (SOD), glutathione (GSH), catalase (CAT), and malondialdehyFde (MDA) were determined using the colorimetric method. Annexin V-FITC and propidium iodide (PI) staining, as well as 33258 were performed using fluorescence microscopy. Moreover, the effect of adding SB203580 was studied to determine the changes in cell apoptosis induced by CPP treatment and Aβ25-35 induction. The CPP markedly inhibited Aβ25-35-induced reduction in the viability and apoptosis of PC12 cells. CPP also reduced the Aβ25-35-induced increase in the expression of the apoptosis factors and the levels of free radicals (ROS and MDA) and reversed the Aβ25-35-induced suppression of antioxidant activity. Additionally, inhibition of p38MAPK via the addition of their antagonists reversed the observed anti-apoptosis effects of CPP. CPP can efficiently provide neuroprotection against Aβ25-35-induced damage in PC12 cells brought about via oxidation and apoptosis reactions, and the underlying mechanisms involve the p38MAPK pathways. Therefore, CPP could potentially be useful as a neuroprotective agent in natural medicine, pharmacy, and the food industry.

Oxidative Challenges Do Not Impact Pheomelanin-Dependent Coloration in Male Japanese Quails.

Colorful traits play an important role in animal communication. Melanin-based colorations are the most extended color traits in animals and are produced by two types of endogenous melanic pigments: eumelanins and pheomelanins, the last ones being the least studied in the context of communication. The production of pheomelanin requires a semi-essential amino acid, cysteine, which is also used for the synthesis of an important endogenous antioxidant, glutathione. Hence, it has been proposed that the synthesis of pheomelanin and glutathione may compete for the cysteine available in the organism. In that case, pheomelanic colorations are predicted to be less intense when the individual is facing an oxidative challenge, and therefore, they would provide information on the oxidative status of the bearer. Here, we experimentally evaluated this hypothesis using male Japanese quails (Coturnix japonica) as a model of study, a species with pheomelanin-based plumage in the breast and cheeks. During feather growth, individuals were exposed to one of three possible conditions: Control (saline), an endogenous oxidative challenge (Escherichia coli lipopolysaccharide injections), or an exogenous oxidative challenge (paraquat injections). Contrary to predictions, we found that: (1) Birds from the three groups exhibited less intense pheomelanic colorations in feathers after the experimental manipulation, and the magnitude of this change did not differ among groups. (2) There was no effect of the experimental treatments on the proportion reduced/oxidized glutathione, an index of oxidative status. (3) Lipid peroxidation was lower after the experimental manipulation, with birds exposed to the paraquat challenge experiencing a stronger decline than other groups. (4) Cysteine and total glutathione levels decreased after the experimental manipulation, with no differences per group in the magnitude of the decline. Taken together the results do not support the hypothesis that oxidative status plays a key role at determining the variation in the intensity of pheomelanic colorations.

Sinapic-Acid-Loaded Nanoparticles Optimized via Experimental Design Methods: Cytotoxic, Antiapoptotoic, Antiproliferative, and Antioxidant Activity.

Nanoparticles are frequently investigated as carrier systems that increase the biological activities of hydrophobic molecules, especially by providing them with water solubility. Sinapic acid (Sa), commonly found in plants, is a phenolic compound with a wide spectrum of biological activities and extensive pharmacological properties. The aim of this study was the synthesis/characterization of optimized sinapic-acid-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (SaNPs) to improve the solubility of sinapic acid (Sa) that limit its use in the biological system and investigate the biological activities of these nanoparticles in the breast cancer cell line. For this purpose, sinapic-acid-loaded PLGA nanoparticles were obtained and optimized by experimental design methods. Then, cytotoxic (MTT method), antiapoptotic (terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay), antiproliferative (immunocytochemically by PCNA assay), and antioxidant activities (superoxide dismutase (SOD) and catalase activities, glutathione, malondialdehyde (MDA), and caspase-3 levels) of optimized nanoparticles were examined comperatively with free drug on MCF-7 cells. The IC50 values of the SaNPs (170.6 ± 3.6 nm size) in MCF-7 cells were determined at 180, 168, and 145 μg/mL for 24, 48, and 72 h, respectively, and at these doses, the nanoparticles did not show any toxic effect on the MCF10A cell line. Treatment of Sa and SaNPs at doses of 24 and 48 h showed a statistically significant reduction in the PCNA level in MCF-7 cells, with an increase in the number of cells leading to apoptosis. In MCF-7 cells treated with SaNP at concentrations of 150 and 200 μg/mL for 24 h, MDA levels were significantly increased, SOD activities were significantly decreased, and reduced glutathione (GSH) and catalase levels were increased compared with control groups. The findings of this study indicate that polyphenolic compounds can contribute to the design of drugs for treatment by forming nanoparticle formulations. The developed nanoparticle formulation is thought to be a useful model for other hydrophobic biological active substances.