Lipid Peroxidation Products Research Articles

The toxicity of superparamagnetic iron oxide nanoparticles induced on the testicular cells: In vitro study

Superparamagnetic iron oxide nanoparticles (SPIONs) have gained significant attention in biomedical research due to their potential applications. However, little is known about their impact and toxicity on testicular cells. To address this issue, we conducted an in vitro study using primary mouse testicular cells, testis fragments, and sperm to investigate the cytotoxic effects of sodium citrate-coated SPIONs (Cit_SPIONs). Herein, we synthesized and physiochemically characterized the Cit_SPIONs and observed that the sodium citrate diminished the size and improved the stability of nanoparticles in solution during the experimental time. The sodium citrate (measured by thermogravimetry) was biocompatible with testicular cells at the used concentration (3%). Despite these favorable physicochemical properties, the in vitro experiments demonstrated the cytotoxicity of Cit_SPIONs, particularly towards testicular somatic cells and sperm cells. Transmission electron microscopy analysis confirmed that Leydig cells preferentially internalized Cit_SPIONs in the organotypic culture system, which resulted in alterations in their cytoplasmic size. Additionally, we found that Cit_SPIONs exposure had detrimental effects on various parameters of sperm cells, including motility, viability, DNA integrity, mitochondrial activity, lipid peroxidation (LPO), and ROS production. Our findings suggest that testicular somatic cells and sperm cells are highly sensitive and vulnerable to Cit_SPIONs and induced oxidative stress. This study emphasizes the potential toxicity of SPIONs, indicating significant threats to the male reproductive system. Our findings highlight the need for detailed development of iron oxide nanoparticles to enhance reproductive nanosafety.

Plant acclimation to ionising radiation requires activation of a detoxification pathway against carbonyl-containing lipid oxidation products.

Ionising γ radiation produces reactive oxygen species by water radiolysis, providing an interesting model approach for studying oxidative stress in plants. Three-week old plants of Arabidopsis thaliana were exposed to a low dose rate (25 mGy h-1) of γ radiation for up to 21 days. This treatment had no effect on plant growth and morphology, but it induced chronic oxidation of lipids which was associated with an accumulation of reactive carbonyl species (RCS). However, contrary to lipid peroxidation, lipid RCS accumulation was transient only, being maximal after 1 day of irradiation and decreasing back to the initial level during the subsequent days of continuous irradiation. This indicates the induction of a carbonyl-metabolising process during chronic ionising radiation. Accordingly, the γ-radiation treatment induced the expression of xenobiotic detoxification-related genes (AER, SDR1, SDR3, ALDH4, and ANAC102). The transcriptomic response of some of those genes (AER, SDR1, and ANAC102) was deregulated in the tga256 mutant affected in three TGAII transcription factors, leading to enhanced and/or prolonged accumulation of RCS and to a marked inhibition of plant growth during irradiation compared to the wild type. These results show that Arabidopsis is able to acclimate to chronic oxidative stress and that this phenomenon requires activation of a carbonyl detoxification mechanism controlled by TGAII. This acclimation did not occur when plants were exposed to an acute γ radiation stress (100 Gy) which led to persistent accumulation of RCS and marked inhibition of plant growth. This study shows the role of secondary products of lipid peroxidation in the detrimental effects of reactive oxygen species.

Ferroptosis inhibitor improves outcome after early and delayed treatment in mild spinal cord injury

We show that redox active iron can induce a regulated form of non-apoptotic cell death and tissue damage called ferroptosis that can contribute to secondary damage and functional loss in the acute and chronic periods after spinal cord injury (SCI) in young, adult, female mice. Phagocytosis of red blood cells at sites of hemorrhage is the main source of iron derived from hemoglobin after SCI. Expression of hemeoxygenase-1 that induces release of iron from heme, is increased in spinal cord macrophages 7 days after injury. While iron is stored safely in ferritin in the injured spinal cord, it can, however, be released by NCOA4-mediated shuttling of ferritin to autophagosomes for degradation (ferritinophagy). This leads to the release of redox active iron that can cause free radical damage. Expression of NCOA4 is increased after SCI, mainly in macrophages. Increase in the ratio of redox active ferrous (Fe2+) to ferric iron (Fe3+) is also detected after SCI by capillary electrophoresis inductively coupled mass spectrometry. These changes are accompanied by other hallmarks of ferroptosis, i.e., deficiency in various elements of the antioxidant glutathione (GSH) pathway. We also detect increases in enzymes that repair membrane lipids (ACSL4 and LPCAT3) and thus promote on-going ferroptosis. These changes are associated with increased levels of 4-hydroxynonenal (4-HNE), a toxic lipid peroxidation product. Mice with mild SCI (30 kdyne force) treated with the ferroptosis inhibitor (UAMC-3203-HCL) either early or delayed times after injury showed improvement in locomotor recovery and secondary damage. Cerebrospinal fluid and serum samples from human SCI cases show evidence of increased iron storage (ferritin), and other iron related molecules, and reduction in GSH. Collectively, these data suggest that ferroptosis contributes to secondary damage after SCI and highlights the possible use of ferroptosis inhibitors to treat SCI.

Green tea supplementation prevented oxidative stress, fibrosis, and myocardial damage in isoproterenol-induced Swiss albino mice

IntroductionOne of the main causes of death and morbidity in people with cardiovascular disorders is myocardial infarction. This study investigated the impact of green tea (GT) leaf powder on isoproterenol (ISO)-induced myocardial infarction in Swiss albino mice. Methods and materials4 to 5 weeks male Swiss albino mice were divided into four groups with 6 mice in every group: Control, ISO, Control + GT and ISO+ GT leaves powder. All of the mice were sacrificed at the end of the investigation. Heart organ and blood plasma samples were taken, and they were tested for oxidative stress indicators and several biochemical parameters. To investigate the formation of mononuclear cells and fibrosis in the heart histopathological staining of tissue sections was also carried out. ResultsIn this experiment, supplementation with green tea leaf powder in ISO-administered mice prevented the increased activity of the AST, ALT, and ALP enzymes. Additionally, three more tests were performed such as CK-MB, Creatinine, and Uric acid whose levels were increased by isoproterenol, and treatment of green tea reduced these elevated levels. Due to the supplementation of green tea leaf powder, ISO-administered mice had lower levels of lipid peroxidation product, nitric oxide, advanced protein oxidation product, and myeloperoxidase. Mice exposed to ISO had considerably higher levels of oxidative stress markers and lower levels of cellular antioxidants including catalase activity, SOD activity, and reduced glutathione concentration. Additionally, the heart of ISO-induced mice revealed substantial fibrosis and inflammatory cell infiltration. Furthermore, green tea leaf powder supplementation prevented inflammatory cell infiltration and fibrosis in ISO-administered mice. ConclusionIn conclusion, the findings imply that supplementing with green tea leaf powder might lower the risk of myocardial infarction in mice given ISO, potentially by reducing oxidative stress, inflammation, and fibrosis.

The Role of Lipid Peroxidation Products in the Development of Oral Sub Mucous Fibrosis- A Cross-Sectional Survey

The Role of Lipid Peroxidation Products in the Development of Oral Sub Mucous Fibrosis- A Cross-Sectional Survey

Supplementation of sperm cryopreservation media with H2S donors enhances sperm quality, reduces oxidative stress, and improves in vitro fertilization outcomes

Cryopreservation of sperm can cause oxidative stress and damage, leading to decreased different functional parameters and fertilization potential. In this study, we evaluated two types of H2S donors: NaHS, a fast-releasing donor, and GYY4137, a slow-releasing donor during cryopreservation of goat sperm. Initially, we determined that 1.5 and 3 μM NaHS, and 15 and 30 μM GYY4137 are optimal concentrations that improved different sperm functional parameters including motility, viability, membrane integrity, lipid peroxidation, and ROS production during incubation at 38.5 °C for 90 min. We subsequently evaluated the impact of the optimal concentration of NaHS and GYY4137 supplementation on various functional parameters following thawing during cryopreservation. Our data revealed that supplementation of extender improved different parameters including post-thaw sperm motility, viability, membrane integrity, and reduced DNA damage compared to the frozen-thawed control group. The supplementation also restored the redox state, decreased lipid peroxidation, and improved mitochondrial membrane potential in the thawed sperm. Finally, we found that supplementation of the extender with NaHS and GYY4137 enhanced IVF outcomes in terms of blastocyst rate and quality of blastocysts. Our results suggest that both donors can be applied for cryopreservation as antioxidants to improve sperm quality and IVF outcomes of frozen-thawed goat sperm.

Regulation of Betaine Homocysteine Methyltransferase by Liver Receptor Homolog-1 in the Methionine Cycle.

Betaine-homocysteine S-methyltransferase (BHMT) is one of the most abundant proteins in the liver and regulates homocysteine metabolism. However, the molecular mechanisms underlying Bhmt transcription have not yet been elucidated. This study aimed to assess the molecular mechanisms underlying Bhmt transcription and the effect of BHMT deficiency on metabolic functions in the liver mediated by liver receptor homolog-1 (LRH-1). During fasting, both Bhmt and Lrh-1 expression increased in the liver of Lrh-1f/f mice; however, Bhmt expression was decreased in LRH-1 liver specific knockout mice. Promoter activity analysis confirmed that LRH-1 binds to a specific site in the Bhmt promoter region. LRH-1 deficiency was associated with elevated production of reactive oxygen species (ROS), lipid peroxidation, and mitochondrial stress in hepatocytes, contributing to hepatic triglyceride (TG) accumulation. In conclusion, this study suggests that the absence of an LRH-1-mediated decrease in Bhmt expression promotes TG accumulation by increasing ROS levels and inducing mitochondrial stress. Therefore, LRH-1 deficiency not only leads to excess ROS production and mitochondrial stress in hepatocytes, but also disrupts the methionine cycle. Understanding these regulatory pathways may pave the way for novel therapeutic interventions against metabolic disorders associated with hepatic lipid accumulation.

#1738 Renal endogenous hepcidin alleviates ferroptosis of collecting duct in ischemia/reperfusion induced AKI

Abstract Background and Aims Recent studies have demonstrated that urinary and serum hepcidin predicted the incidence of acute kidney injury (AKI) among critically ill patients. As an endogenous acute phase hormone, hepcidin is synthesized not only by the liver but also secreted by distal nephron. However, the autocrine impact of hepcidin secreted by distal nephron on collecting duct cells in AKI remains poorly understood. Method We determined the expression and location of hepcidin in murine kidney in different severity of bilateral ischemia/reperfusion (bIRI) induced AKI model with ischemic duration for 20 min and 32 min. The effect of renal endogenous hepcidin was explored in cultured M-1 (Murine Cortical collecting duct) exposed to hypoxia/reoxygenation (H/R), as well as in bIRI mice with collecting duct-specific overexpression of hepcidin. Transgenic mice were generated by genetic cross between Hepcidin floxed mice which knocked the gene fragment of CAG-LSL-mHamp-flag-WPRE-PolyA into H11 locus and Cdh16-cre mice. Mechanistically, RNA-seq was performed in M-1 cells under H/R condition. Results The expression of renal endogenous hepcidin was significantly elevated in bIRI32min mice compared to both bIRI20min and sham group, suggesting that augmented secretion of hepcidin was induced by more severe AKI. Interestingly, renal endogenous hepcidin was mainly localized to distal convoluted tubule and collecting tubule as indicated by its co-staining with AQP2 and Calbindin-D28k in the kidney. To investigate the role of hepcidin in collecting duct, M-1 cells were cultured under hypoxia condition. We observed remarkable ferroptosis in collecting duct cells other than apoptosis nor necroptosis. Subsequently, hepcidin knockdown further exacerbated ferroptosis under H/R condition, as evidenced by an upregulation of PTGS2 expression and downregulation of GXP4 expression, lower proportion of GSH/GSSG ratio as well as enhanced lipid peroxide accumulation and production of Fe2+. Then, RNA-seq data revealed that hepcidin knockdown promoted the enrichment of genes related to oxidative stress and glutathione metabolic pathway which may be responsible for the deterioration of ferroptosis. To investigate the role of hepcidin in vivo, bIRI induced AKI was established in Cdh16-cre+Hepcidinflox/+ mice. We observed remarkable overexpression of Hepcidin-Flag localized to collecting duct and interstium area. Impressively, Cdh16-cre+Hepcidinflox/+ mice showed ameliorated renal dysfunction adrenal tubule injury histologically. Ferroptosis of renal was alleviated in Cdh16-cre+Hepcidinflox/+ mice as confirmed by an upregulation of GPX4 expression and downregulation of PTGS2, ACSL4, FPN expression, higher proportion of GSH/GSSG ratio, as well as reduced lipid peroxide accumulation and production of Fe2+ by Lipofluo (lipid peroxides) and FerroOrange (Fe2+) staining on frozen section of kidney. Ferroptosis of the collecting duct was alleviated as indicated by downregulation of PTGS2 expression which co-staining with AQP2. To demonstrate the beneficial role of hepcidin, exogenous hepcidin was applied to bIRI induced AKI mice to strengthen the endogenous protective response. Conclusion Renal endogenous hepcidin is induced in the distal tubule and collecting duct after AKI as a protective response via alleviating ferroptosis of collecting duct. Hepcidin may represent a novel therapeutic approach to promote tubule repair by targeting ferroptosis for AKI.

#999 PDK4 triggers ferroptosis in diabetic kidney disease by promoting NRF2 ubiquitination

Abstract Background and Aims Diabetic kidney disease (DKD) is the principal cause of chronic kidney disease and end-stage renal disease. Despite this burden, the detailed understanding of the pathogenic mechanism that promote the initiation and progression of DKD remain to be fully elucidated. This study aims to investigate the role of pyruvate dehydrogenase kinase 4 (PDK4) in diabetic renal tubular cell death from the perspectives of mitochondrial injury, metabolic reprogramming and ferroptosis. Method Cellular transcriptomics was employed to screen the differential genes between normal glucose (as control) and 35 mM glucose combined with 120 uM palmitic acid (as HGL) groups. The vitro experimental groups include Ctl, HGL, HGL/DCA (5 mM PDK4 inhibitor, sodium dichloroacetate), HGL/DMSO, culturing for 48 h. C57BL/6J mice were used to perform the vivo experiments. A total of 40 C57BL/6J mice were randomly divided into 5 groups respectively labeled as control, DKD, DKD/DCA (25 mg/kg, daily), DKD/DMSO (1% DMSO, daily) and DKD/Fer-1 (1 mg/kg, daily) group. Western blot, qRT-PCR, Si-RNA transfection, transmission electron microscopy, ROS probe, FerroOrange/Green probes, liperfluo probe, immunofluorescence and Co-IP assay were used in this study to detect the indicators of metabolic reprogramming, mitochondrial function, oxidative stress and ferroptosis, as well as the NRF2 ubiquitination level. Results Based on the cellular transcriptomics, PDK4 mRNA was significantly elevated in the HK-2 cells treated with HGL. Likewise, mRNA and protein levels were also elevated in the kidneys of mice treated with streptozotocin and high fat diet. In kidneys with PDK4 inhibitor DCA treatment or PDK4 genetic knockdown, glycolipid toxicity-induced metabolic reprogramming and destruction of mitochondrial function were markedly ameliorated. In addition, mitochondrial reactive oxygen species and the oxidative stress markers NADPH oxidase 2 and 4 (NOX2, NOX4) were diminished in DCA-treated or PDK4 knockdown HK-2 cells, while superoxide dismutase 1 and 2 (SOD1, SOD2) and catalase (CAT) expression and glutathione (GSH) levels were restored. DCA intervention or PDK4 knockdown in DKD mice or HK-2 cells alleviated the signs of ferroptosis caused by glycolipid toxicity, demonstrated as up-regulation of SLCA11, GPX4, FSP1 and GSH to maintain the balance of oxidative stress, increase of FTH1 and FPN1 to promote Fe2+ storage and discharge, decrease of TFR1 to reduce Fe2+ intake, as well as reduction of lipid peroxidation products including 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA). PDK4 promoted NRF2 ubiquitination under HGL condition, since inhibition or knockdown of PDK4 in HK-2 cells increased the protein level of NRF2, while the mRNA level was unchanged. Conclusion Collectively, we identified a new ferroptosis inducer gene, PDK4, which is commonly up-regulated in diabetes. PDK4-induced metabolic reprogramming, mitochondrial damage and oxidative stress promote ROS accumulation and ultimately lead to ferroptosis in DKD, this effect can be achieved in part by promoting NRF2 ubiquitination.

The beneficial effect of fluoxetine on behavioral and cognitive changes in chronic experimental Chagas disease unveils the role of serotonin fueling astrocyte infection by Trypanosoma cruzi.

In Chagas disease (CD), a neglected tropical disease caused by the parasite Trypanosoma cruzi, the development of mental disorders such as anxiety, depression, and memory loss may be underpinned by social, psychological, and biological stressors. Here, we investigated biological factors underlying behavioral changes in a preclinical model of CD. In T. cruzi-infected C57BL/6 mice, a kinetic study (5 to 150 days postinfection, dpi) using standardized methods revealed a sequential onset of behavioral changes: reduced innate compulsive behavior, followed by anxiety and depressive-like behavior, ending with progressive memory impairments. Hence, T. cruzi-infected mice were treated (120 to 150 dpi) with 10 mg/Kg/day of the selective serotonin reuptake inhibitor fluoxetine (Fx), an antidepressant that favors neuroplasticity. Fx therapy reversed the innate compulsive behavior loss, anxiety, and depressive-like behavior while preventing or reversing memory deficits. Biochemical, histological, and parasitological analyses of the brain tissue showed increased levels of the neurotransmitters GABA/glutamate and lipid peroxidation products and decreased expression of brain-derived neurotrophic factor in the absence of neuroinflammation at 150 dpi. Fx therapy ameliorated the neurochemical changes and reduced parasite load in the brain tissue. Next, using the human U-87 MG astroglioma cell line, we found no direct effect of Fx on parasite load. Crucially, serotonin/5-HT (Ser/5-HT) promoted parasite uptake, an effect increased by prior stimulation with IFNγ and TNF but abrogated by Fx. Also, Fx blocked the cytokine-driven Ser/5-HT-promoted increase of nitric oxide and glutamate levels in infected cells. We bring the first evidence of a sequential onset of behavioral changes in T. cruzi-infected mice. Fx therapy improves behavioral and biological changes and parasite control in the brain tissue. Moreover, in the central nervous system, cytokine-driven Ser/5-HT consumption may favor parasite persistence, disrupting neurotransmitter balance and promoting a neurotoxic environment likely contributing to behavioral and cognitive disorders.

Highly selective and effective ferroptosis liposomal nanodrugs for synergistic antitumor therapy

Ferroptosis is a recently discovered emerging regulated cell death pattern characterized by iron-dependent lipid peroxidation and has enormous potential in tumor therapy. However, the antioxidant defense system during tumor cell metabolism hampers the ferroptosis-mediated anticancer effect, especially the GSH/GPX4 pathway, which is regarded as the classic means of scavenging excessive lipid peroxidation production. Here, we developed an iron-based nanodrug (ipFFR) with efficient Fenton catalytic activity and encapsulated the ferroptosis inducer RSL3. This formulation consists of Fe@Fe3O4 nanoparticles and RSL3, encapsulated in nanoliposomes containing DSPE-PEG-iRGD for precise tumor-targeted delivery of ipFFR. The ultrasmall Fe@Fe3O4 metal nanoparticles were enabled to catalyze endogenous H2O2 and produce toxic •OH under weakly acidic condition and were then quickly excreted from the body. Furthermore, the released ferroptosis inducer RSL3 specifically inhibited the activity of glutathione peroxidase 4 (GPX4) to block the antioxidant pathway of ferroptosis, synergizing and thereby enhancing ferroptosis therapy. This study may open up new avenues in designing nanomedicines with synergistic effects to enhance ferroptosis therapy.

Intervention effects of low-molecular-weight chondroitin sulfate from the nasal cartilage of yellow cattle on lipopolysaccharide-induced behavioral disorders: regulation of the microbiome-gut-brain axis.

Chondroitin sulfate (CS) is a sulfated linear polysaccharide with different functional activities, including antioxidant, anti-inflammatory, lipid-lowering, and immune regulation. As natural sulfated polysaccharides have high molecular weight, high apparent viscosity, low water solubility, complex structure, and high negative charge, they have difficulty binding to receptors within cells across tissue barriers, resulting in low bioavailability and unclear structure-activity relationships. In this study, an H2O2-Vc oxidative degradation system was employed to perform environmentally friendly and controllable degradation of CS extracted from the nasal cartilage of Shaanxi Yellow cattle. Two low-molecular-weight chondroitin sulfates (LMWCSs), CS-1 (14.8 kDa) and CS-2 (50.9 kDa), that exhibit strong in vitro free radical scavenging ability were obtained, and their structures were characterized. Mice intraperitoneally administered lipopolysaccharide (LPS) were used to explore the cognitive intervention effects of LMWCS. Supplementing CS-1 and CS-2 significantly downregulated the levels of the serum inflammatory factors, TNF-α and IL-1β, promoted the expression of GSH in the brain, and inhibited the production of the lipid peroxidation product, malondialdehyde (MDA), ultimately inhibiting LPS-induced cognitive impairment in mice. Surprisingly, compared to the LPS model group, the abundances of Streptococcus, Eisenbergiella, Vampirovibrio, Coprococcus, Enterococcus and Lachnoanaerobaculum were significantly increased in the intestines of mice in the CS-1 and CS-2 group, whereas those of Parabacteroides and Mycoplasma were significantly decreased. Altogether, this study provides a theoretical basis for the comprehensive utilization of agricultural and animal resources and the application of brain nutrition, anti-inflammatory, and LMWCS health products.

Sodium-glucose cotransporter 2 inhibitor dapagliflozin prevents ejection fraction reduction, reduces myocardial and renal NF-κB expression and systemic pro-inflammatory biomarkers in models of short-term doxorubicin cardiotoxicity.

Anthracycline-mediated adverse cardiovascular events are among the leading causes of morbidity and mortality in patients with cancer. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) exert multiple cardiometabolic benefits in patients with/without type 2 diabetes, chronic kidney disease, and heart failure with reduced and preserved ejection fraction. We hypothesized that the SGLT2i dapagliflozin administered before and during doxorubicin (DOXO) therapy could prevent cardiac dysfunction and reduce pro-inflammatory pathways in preclinical models. Cardiomyocytes were exposed to DOXO alone or combined with dapagliflozin (DAPA) at 10 and 100 nM for 24 h; cell viability, iATP, and Ca++ were quantified; lipid peroxidation products (malondialdehyde and 4-hydroxy 2-hexenal), NLRP3, MyD88, and cytokines were also analyzed through selective colorimetric and enzyme-linked immunosorbent assay (ELISA) methods. Female C57Bl/6 mice were treated for 10 days with a saline solution or DOXO (2.17 mg/kg), DAPA (10 mg/kg), or DOXO combined with DAPA. Systemic levels of ferroptosis-related biomarkers, galectin-3, high-sensitivity C-reactive protein (hs-CRP), and pro-inflammatory chemokines (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL17-α, IL-18, IFN-γ, TNF-α, G-CSF, and GM-CSF) were quantified. After treatments, immunohistochemical staining of myocardial and renal p65/NF-kB was performed. DAPA exerts cytoprotective, antioxidant, and anti-inflammatory properties in human cardiomyocytes exposed to DOXO by reducing iATP and iCa++ levels, lipid peroxidation, NLRP-3, and MyD88 expression. Pro-inflammatory intracellular cytokines were also reduced. In preclinical models, DAPA prevented the reduction of radial and longitudinal strain and ejection fraction after 10 days of treatment with DOXO. A reduced myocardial expression of NLRP-3 and MyD-88 was seen in the DOXO-DAPA group compared to DOXO mice. Systemic levels of IL-1β, IL-6, TNF-α, G-CSF, and GM-CSF were significantly reduced after treatment with DAPA. Serum levels of galectine-3 and hs-CRP were strongly enhanced in the DOXO group; on the other hand, their expression was reduced in the DAPA-DOXO group. Troponin-T, B-type natriuretic peptide (BNP), and N-Terminal Pro-BNP (NT-pro-BNP) were strongly reduced in the DOXO-DAPA group, revealing cardioprotective properties of SGLT2i. Mice treated with DOXO and DAPA exhibited reduced myocardial and renal NF-kB expression. The overall picture of the study encourages the use of DAPA in the primary prevention of cardiomyopathies induced by anthracyclines in patients with cancer.

About Lipid Peroxidation During Physical Activity

In recent years, interest in the study of free radical processes occurring in living organisms has increased significantly. The study of free radical processes, in particular lipid peroxidation, lipid activity, and the activity of enzymes involved in reactions, is of interest for identifying the characteristics of functional activity. Lipid peroxidation products are considered as biological destructive factors, the accumulation of which in the body induces oxidative stress. As a result, the features of the process of lipid peroxidation and the antioxidant protective system of muscle and other tissues that provide motor activity were identified. The results can be used in physical education classes.

Use of coenzyme Q-10 to improve the pregnancy rate in sheep

One of the factors responsible for less pregnancy rates is the use of frozen semen in sheep due to the oxidative stress created by the process. The aim of this experiment was to test the effects of adding coenzyme Q-10 (CoQ10) to the seminal extender on sperm quality and the pregnancy rate of sheep. In this study, ejaculates from eight Dorper rams of reproductive age were used and tested in four treatments: Control (pure BotuBov®), C1 (175 µM of CoQ10), C3 (350 µM of CoQ10), and C7 (700 µM of CoQ10). Samples were collected in triplicate from each animal, and sperm analysis was performed by CASA after thawing at 0 h and 2 h. The samples were also analyzed by flow cytometry for plasma and acrosomal membrane integrity, stability, lipid peroxidation, mitochondrial potential, and superoxide anion production. In total, 198 ewes were inseminated by laparoscopy and divided into two groups: control (n=98) and C7 (n=100). Pregnancy diagnosis was performed at 30 days. Coenzyme Q10 proved to be safe for semen cryopreservation, not altering sperm kinetic values between the groups post-thawing. In flow cytometry, the C1 and C7 groups achieved a better index of plasma membrane integrity and membrane stability (P<0.05). A increased pregnancy rate was observed in C7 (52 %) compared to the control (38 %). In conclusion, coenzyme Q10 assists in the cryopreservation process, protecting the sperm cell and improving pregnancy rates in ewes.

A dual model of normal vs isogenic Nrf2-depleted murine epithelial cells to explore oxidative stress involvement

Cancer-derived cell lines are useful tools for studying cellular metabolism and xenobiotic toxicity, but they are not suitable for modeling the biological effects of food contaminants or natural biomolecules on healthy colonic epithelial cells in a normal genetic context. The toxicological properties of such compounds may rely on their oxidative properties. Therefore, it appears to be necessary to develop a dual-cell model in a normal genetic context that allows to define the importance of oxidative stress in the observed toxicity. Given that the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) is considered to be the master regulator of antioxidant defenses, our aim was to develop a cellular model comparing normal and Nrf2-depleted isogenic cells to qualify oxidative stress–related toxicity. We generated these cells by using the CRISPR/Cas9 technique. Whole-genome sequencing enabled us to confirm that our cell lines were free of cancer-related mutations. We used 4-hydroxy-2-nonenal (HNE), a lipid peroxidation product closely related to oxidative stress, as a model molecule. Here we report significant differences between the two cell lines in glutathione levels, gene regulation, and cell viability after HNE treatment. The results support the ability of our dual-cell model to study the role of oxidative stress in xenobiotic toxicity.

The influence of “Butaintersyl” on the antioxidant status of rats under conditions of toxic damage caused by tetrachloromethane

Tetrachloromethane (CCl4) is a toxic chemical substance known in scientific research as a model for studying the damage of parenchymal liver cells. The mechanisms of CCl4 toxicity include the activation of lipid peroxidation processes, the intensive formation of free radicals, and, as a result, the disruption of the pro-/antioxidant balance. This work aimed to study the effect of the liposomal drug “Butaintersyl” on the activity of the glutathione antioxidant protection system and the intensity of lipid peroxidation processes in the blood of rats under conditions of toxic damage caused by tetrachloromethane. The studies were conducted on white sexually mature young Wistar rats weighing 180–200 g, kept in the institute's vivarium of the State Scientific Research Control Institute of Veterinary Drugs and Feed Additives. In the blood of the rats, changes in the activity of glutathione peroxidase and the level of reduced glutathione were studied, as well as the levels of lipid peroxidation products: lipid hydroperoxides and TBA-active products. The development of oxidative stress in the first experimental group of rats was accompanied by the suppression of the glutathione antioxidant protection system, as evidenced by a decrease in blood glutathione peroxidase activity and the level of reduced glutathione. At the same time, the intoxicated animals showed an increase in lipid peroxidation processes, namely an increase in the blood levels of lipid hydroperoxides and TBA-active products throughout the study period. The studies showed that in cases of poisoning of various origins, it is advisable to use drugs that reduce the formation of reactive oxygen species and lipoperoxidation processes, exhibit antioxidant effects, and stabilize cell membranes. A special place among such drugs is occupied by the liposomal drug “Butaintersyl”. This drug ensures the stabilization of biological membranes. The liposomal drug “Butaintersyl” contributed to suppressing lipid peroxidation processes and activating the antioxidant protection system, as evidenced by the high content of reduced glutathione and glutathione peroxidase activity. Due to its antioxidant properties, the drug positively affected the activity of membrane-dependent enzymes. It reduced the level of endogenous intoxication, allowing it to be recommended for inclusion in prevention schemes for toxic liver damage caused by chemical compounds.

Enhancing nutrient digestibility and antioxidant efficacy in piglets: The impact of fermented rapeseed meal supplementation on biochemical parameters and oxidative stress markers

Abstract This study aimed to investigate the effects of varying levels of dried fermented rapeseed meal (FRSM) on the nutrient digestibility and antioxidant efficacy in piglets. The experiment was conducted on 300 piglets, starting at 18 days of age. The primary experimental factor was the inclusion of FRSM in the diet, substituting mainly for post-extraction soybean meal and wheat. Two control groups were established: a positive control (PC) supplemented with 2500 ppm zinc oxide, and a negative control (NC) without additives. Additionally, four experimental groups were designated based on FRSM inclusion levels: FR-8 (8% FRSM), FR-12 (12% FRSM), FR-15 (15% FRSM), and FR-25 (25% FRSM). Results indicated a positive impact on nutrient absorption and redox processes, characterized by reduced efficiency of lipid peroxidation products such as malondialdehyde and lipid hydroperoxide in both blood and organs like the liver and intestines. From this perspective, the recommended inclusion of fermented rapeseed meal in piglet feed appears to be within the range of 8–12%

Qiangxinyin formula protects against isoproterenol-induced cardiac hypertrophy

Heart failure is a life-threatening cardiovascular disease and characterized by cardiac hypertrophy, inflammation and fibrosis. The traditional Chinese medicine formula Qiangxinyin (QXY) is effective for the treatment of heart failure while the underlying mechanism is not clear. This study aims to identify the active ingredients of QXY and explore its mechanisms protecting against cardiac hypertrophy. We found that QXY significantly protected against isoproterenol (ISO)-induced cardiac hypertrophy and dysfunction in zebrafish. Eight compounds, including benzoylmesaconine (BMA), atractylenolide I (ATL I), icariin (ICA), quercitrin (QUE), psoralen (PRN), kaempferol (KMP), ferulic acid (FA) and protocatechuic acid (PCA) were identified from QXY. PRN, KMP and icaritin (ICT), an active pharmaceutical ingredient of ICA, prevented ISO-induced cardiac hypertrophy and dysfunction in zebrafish. In H9c2 cardiomyocyte treated with ISO, QXY significantly blocked the calcium influx, reduced intracellular lipid peroxidative product MDA, stimulated ATP production and increased mitochondrial membrane potential. QXY also inhibited ISO-induced cardiomyocyte hypertrophy and cytoskeleton reorganization. Mechanistically, QXY enhanced the phosphorylation of Smad family member 2 (SMAD2) and myosin phosphatase target subunit-1 (MYPT1), and suppressed the phosphorylation of myosin light chain (MLC). In conclusion, PRN, KMP and ICA are the main active ingredients of QXY that protect against ISO-induced cardiac hypertrophy and dysfunction largely via the blockage of calcium influx and inhibition of mitochondrial dysfunction as well as cytoskeleton reorganization.

α-Tocopherol inhibits atherogenesis and improves cardiac function in mice independently of its antioxidant properties.

The impact of α-tocopherol on atherosclerosis is unclear and controversial. While some studies suggest potential benefits, such as antioxidant properties that may reduce oxidative stress, other studies indicate no significant preventive effects. The intricate interplay of various factors, including dosage, individual differences, and study methodologies, contributes to the ongoing uncertainty surrounding α-tocopherol's role in atherosclerosis. Further research is needed to clarify its impact and establish clearer guidelines. Therefore, we aimed to evaluate the impact of α-tocopherol on atherogenesis in ApoE-/- fibrillin (Fbn)1C1039G/+ mice, which is a unique mouse model of advanced atherosclerosis with typical features, such as large necrotic cores, high levels of inflammation, and intraplaque neovascularization, that resemble the unstable phenotype of human plaques. ApoE-/- Fbn1C1039G+/- mice were fed a western-type diet (WD) supplemented with a high dose of α-tocopherol (500 mg/kg diet), while control mice were fed a WD containing a low dose of α-tocopherol (50 mg/kg diet). The high dose of α-tocopherol reduced plaque thickness and necrotic core area in the right common carotid artery (RCCA) after 24 weeks WD. Moreover, α-tocopherol decreased plaque formation and intraplaque neovascularization in the RCCA. In addition to its antiatherogenic effect, chronic supplementation of α-tocopherol improved cardiac function in ApoE-/- Fbn1C1039G/+ mice. However, chronic supplementation of α-tocopherol did not decrease lipid peroxidation. On the contrary, α-tocopherol acted as a prooxidant by increasing plasma levels of oxidized LDL and plaque malondialdehyde, an end product of lipid peroxidation. Our data indicate that α-tocopherol inhibits atherogenesis and improves cardiac function independent of its antioxidant properties.