Detection Of Lipid Peroxidation Research Articles

Exogenously applied putrescine regulates aluminium [al (III)] stress in maize (Zea mays L.): Physiological and metabolic implications

In this study, we investigated the role of exogenous application of putrescine (Put) for alleviation of Al (III) stress responses in maize seedlings. In presence of Put, the growth and biomass improved substantially in Al (III) stressed seedlings as compared to Al (III) alone. The production of hydrogen peroxide (H2O2) and superoxide radical (O2•-) were significantly restrained in during Al (III) stress in Put supplemented seedlings. The malondialdehyde (MDA) production was significantly controlled during Al (III) stress by Put supplementation as compared to the seedlings under Al (III) stress alone. The histochemical analysis showed variation in staining intensities of diaminobenzidine (DAB) and nitoroblue tetrazolium (NBT) salt to detect the presence of H2O2 and O2•- respectively, reflected strong role of Put in regulation of reactive oxygen species (ROS) production. The staining of roots with Schiff's reagent and Evans blue dye to detect lipid peroxidation and loss of plasma membrane integrity showed that Put controls the lipid peroxidation and maintains the integrity of the plasma membrane during Al (III) stress. The proton (1H) nuclear magnetic resonance (NMR) analysis of methanolic extracts of root and shoot of maize seedlings under Al (III) stress in presence or absence of Put showed considerable disparity in terms of metabolic profiles, which include amino acids, sugars, carbohydrates, organic acids and several aliphatic and aromatic compounds. The present study exhibited the potential role of Put for mitigation of Al (III) stress by enhancing growth, regulating ROS production, reducing oxidative stress, improving antioxidant metabolism and maintaining the metabolic pool.

Iron Overload Induces Hepatic Ferroptosis and Insulin Resistance by Inhibiting the Jak2/stat3/slc7a11 Signaling Pathway.

Recent studies showed that patients with iron overload had increased risk of insulin resistance or diabetes. Ferroptosis is a new type of cell death mainly caused by iron-dependent oxidative damage. In the present study, we investigated potential mechanisms of iron overload induced hepatic ferroptosis and insulin resistance through in vivo and in vitro experiments. In vivo, the mice models of iron overload were established by intraperitoneal injection of iron dextran. The changes of body weight, serum ferritin and blood glucose were measured. Hematoxylin-eosin (HE) and Perl's stainings were used to observe the pathological changes and iron deposition in the liver of mice. In vitro, HepG2 cells were treated with ferric ammonium citrate (FAC, 9 mmol/L, 24 h) to establish the cell models of iron overload. The labile iron pool, cell viability, glucose consumption and glycogen contents were measured. The ultrastructure of mitochondria was observed by transmission electron microscope (TEM). The malondialdehyde (MDA) and glutathione (GSH) kits were used to detect lipid peroxidation in liver tissues of mice and HepG2 cells. RT-PCR and Western blot were used to detect the mRNA and protein expression levels of ferroptosis factors and JAK2/STAT3 signaling pathway. In this study, we used the iron chelator deferasirox in mice and HepG2 cells. Iron overload caused weight loss, elevated serum ferritin, fasting blood glucose, fasting insulin, HOMA-IR, impaired glucose tolerance, and decreased insulin sensitivity in mice. HE staining and Perls staining showed clumps of iron deposition in the liver of iron overload mice. Iron overload could reduce the glucose consumption, increase MDA contents of HepG2 cells, while reduce glycogen and GSH contents in liver tissues of mice and HepG2 cells. TEM showed deletion of mitochondrial ridge and rupture of outer membrane in HepG2 cells with iron overload. Iron chelator deferasirox could significantly improve the above indicators, which might be related to the activation of JAK2/STAT3/SLC7A11 signaling pathway and hepatic ferroptosis. Iron overload could induce hepatic ferroptosis and insulin resistance by inhibiting the JAK2/STAT3/SLC7A11 signaling pathway, and the iron chelator deferasirox might improve hepatic insulin resistance induced by iron overload.

Antibacterial mechanism and enhancement of antibacterial activity under visible light in Er3+-doped BiOBr

The introduction of oxygen defects in semiconductor electronic structures is considered to be an effective strategy to improve their photocatalytic activity. In this work, rare-earth Er3+−doped BiOBr (Er/BiOBr) with oxygen-rich vacancies (OVs) was prepared via simple hydrothermal method. Compared with pure BiOBr, Er/BiOBr has a narrower band gap and more abundant OVs, which together improve the separation efficiency of photogenerated carriers. The introduction of Er not only creates impurity states between conduction band (CB) and valence band (VB), but also produces more OVs as electron traps. Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) were used as evaluation systems for these materials with different Er3+ doping contents. The results showed that under the combined action of ·O2− and H2O2, 2.0% EBB samples with Er3+ doping content of BiOBr showed the best broad-spectrum antibacterial activity, and E. coli and S. aureus were completely inactivated at 0.03 mg ml−1 and 0.05 mg ml−1 concentrations. The surface and interfacial structures of Er/BiOBr samples were studied by XRD, brunauer-emmet-teller method, SEM, TEM and EDX. OVs and photogenerated carrier separation in Er/BiOBr were determined by ESR, PL and photochemical detection methods. More importantly, through the detection of lipid peroxidation and bacterial respiratory chain dehydrogenase activities, the internal and external processes of ROS on bacterial inactivation were determined, and the photocatalytic antibacterial mechanism was summarized. This study for the antimicrobial properties of rare earth ions doped improve photocatalytic materials provides a new train of thought.

Baicalein, a component of banxia xiexin decoction, alleviates CPT-11-induced gastrointestinal dysfunction by inhibiting ALOX15-mediated ferroptosis.

Baicalein, one of the active ingredients of banxia xiexin decoction, has good therapeutic efficacy in treating diarrhea and improving gastrointestinal dysfunction. The role and mechanism of Baicalein on irinotecan (CPT-11)-induced gastrointestinal dysfunction are the focus of this study. Concretely, CPT-11 induced delayed diarrhea rat model and intestinal epithelial cell (IEC)-6 cell injury model with Baicalein treatment as needed. Colonic pathological changes were analyzed by hematoxylin-eosin staining, and inflammatory factor expressions in serum were determined by enzyme-linked immunosorbent assay. Immunohistochemistry and western blot were performed to quantify ferroptosis-related protein expressions. Thiobarbituric acid reactive substances (TBARS) kits and colorimetric assay kit were applied to detect lipid peroxidation levels and Fe2+ content, respectively. In vitro experiments also included quantitative real-time polymerase chain reaction, cell counting kit-8, and C11 BODIPY staining. CPT-11 induced aggravation of intestinal tissue damage, inflammatory factor release, Fe2+ accumulation, upregulation of lipid peroxidation and 15-Lipoxygenase (ALOX15) expression, and downregulation of glutathione peroxidase 4 (Gpx4) and SLC7A11 in vivo in rats; however, Baicalein dose-dependently reversed the effects of CPT-11. Baicalein elevated cell viability, reduced lipid peroxidation and Fe2+ accumulation, and elevated Gpx4 and SLC7A11 levels, whereas ALOX15 overexpression reversed the effects of Baicalein on a CPT-11-induced IEC-6 cell injury model. In conclusion, Baicalein plays a mitigating role in CPT-11-induced delayed diarrhea via ALOX15-mediated ferroptosis.

Cytotoxicity and hemolysis of rare earth ions and nanoscale/bulk oxides (La, Gd, and Yb): Interaction with lipid membranes and protein corona formation

The widespread application of rare earth elements (REEs) has raised concerns about their potential release into the environment and subsequent ingestion by humans. Therefore, it is essential to evaluate the cytotoxicity of REEs. Here, we investigated the interactions between three typical REEs (La, Gd, and Yb) ions as well as their nanometer/μm-sized oxides and red blood cells (RBCs), a plausible contact target for nanoparticles when they enter the bloodstream. Hemolysis of REEs at 50–2000 μmol L−1 was examined to simulate their cytotoxicity under medical or occupational exposure. We found that the hemolysis due to the exposure of REEs was highly dependent on their concentration, and the cytotoxicity followed the order of La3+ > Gd3+ > Yb3+. The cytotoxicity of REE ions (REIs) is higher than REE oxides (REOs), while nanometer-sized REO caused more hemolysis than that μm-sized REO. The production of reactive oxygen species (ROS), ROS quenching experiment, as well as the detection of lipid peroxidation, confirmed that REEs causes cell membrane rupture by ROS-related chemical oxidation. In addition, we found that the formation of a protein corona on REEs increased the steric repulsion between REEs and cell membranes, hence mitigating the cytotoxicity of REEs. The theoretical simulation indicated the favorable interaction of REEs with phospholipids and proteins. Therefore, our findings provide a mechanistic explanation for the cytotoxicity of REEs to RBCs once they have entered the blood circulation system of organisms.

Alpha-Tocopherol from People to Plants Is an Essential Cog in the Metabolic Machinery.

Significance: Protection from oxygen, a diradical, became a necessity with the evolution of photosynthetic organisms about 2.7 billion years. α-Tocopherol plays an essential, protective role in organisms from plants to people. An overview of human conditions that result in severe vitamin E (α-tocopherol) deficiency is provided. Recent Advances: α-Tocopherol has a critical role in the oxygen protection system by stopping lipid peroxidation, its induced damage, and cellular death by ferroptosis. Recent findings in bacteria and plants support the concept of why lipid peroxidation is so dangerous to life and why the family of tocochromanols are essential for aerobic organisms and for plants. Critical Issues: The hypothesis that prevention of the propagation of lipid peroxidation is the basis for the α-tocopherol requirement in vertebrates is proposed and further that its absence dysregulates energy metabolism, one-carbon metabolism, and thiol homeostasis. By recruiting intermediate metabolites from adjacent pathways to sustain effective lipid hydroperoxide elimination, α-tocopherol function is linked not only to NADPH metabolism and its formation through the pentose phosphate pathway via glucose metabolism but also to sulfur-containing amino acid metabolism and to one-carbon metabolism. Future Directions: Evidence from humans, animals, and plants supports the hypothesis, but future studies are needed to assess the genetic sensors that detect lipid peroxidation and cause the ensuing metabolic dysregulation. Antioxid. Redox Signal. 38, 775-791.

Probing Lipid Peroxidation in Ferroptosis: Emphasizing the Utilization of C11-BODIPY-Based Protocols.

Ferroptosis is a form of regulated cell death that relies on iron and is characterized by the accumulation of lipid peroxides, resulting in oncotic cell swelling and eventual disruption of cellular membranes. Lipid peroxidation, a hallmark of ferroptosis, refers to the oxidative deterioration of lipids that contain carbon-carbon double bonds, particularly polyunsaturated fatty acids (PUFAs). Understanding the molecular mechanisms underlying the interplay between ferroptosis and lipid peroxidation and identifying reliable techniques for assessing lipid peroxidation levels are crucial for further advancements in this field of research. Various methods have been developed to detect lipid peroxidation levels, including C11-BODIPY (BODIPY™ 581/591 C11), liperfluo, 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), Click-iT LAA (linoleamide alkyne), and liquid chromatography-mass spectrometry (LC-MS)-based epilipidomics (redox lipidomics). Currently, one of the most commonly used and effective methods is the C11-BODIPY assay, which utilizes a fluorescent probe that selectively sensitizes lipid peroxidation in cell membranes. Incorporating advanced techniques such as flow cytometry and fluorescence microscopy with C11-BODIPY dye is essential for accurate assessment of lipid peroxidation levels in ferroptosis. This chapter aims to provide comprehensive experimental protocols for detecting lipid peroxidation levels indicative of ferroptosis using C11-BODIPY staining and subsequent detection via flow cytometry and fluorescence microscopy.

Growth Differentiation Factor 15 Regulates Oxidative Stress-Dependent Ferroptosis Post Spinal Cord Injury by Stabilizing the p62-Keap1-Nrf2 Signaling Pathway.

BackgroundSpinal cord injury (SCI) is a severe traumatic disorder of the central nervous system (CNS) that causes irreversible damage to the nervous tissue. The consequent hemorrhage contributed by trauma induces neuronal ferroptosis post SCI, which is an important death mode to mediate neuronal loss. Growth differentiation factor 15 (GDF15) is a cytokine that regulates cell proliferation, differentiation, and death. However, the specific role of GDF15 in neuronal ferroptosis post SCI remains unknown.Materials and MethodsNeuronal ferroptosis in vitro was measured by detection of lipid peroxidation, glutathione, iron content, and reactive oxidative stress. In vivo, western blotting and immunofluorescence (IF) staining was utilized to measure ferroptosis post SCI. IF staining, TUNEL staining, hematoxylin-eosin staining, and Nissl staining were used to measure neurological damage. Finally, locomotor function recovery was analyzed using the Basso Mouse Scale and Louisville Swim Scale.ResultsGDF15 was significantly increased in neuronal ferroptosis and silencing GDF15 aggravated ferroptosis both in vitro and in vivo. Besides, GDF15-mediated inhibition of neuronal ferroptosis is through p62-dependent Keap1-Nrf2 pathway. In SCI mice, knockdown of GDF15 significantly exacerbated neuronal death, interfered with axon regeneration and remyelination, aggravated ferroptosis-mediated neuroinflammation, and restrained locomotor recovery.ConclusionGDF15 effectively alleviated neuronal ferroptosis post SCI via the p62-Keap1-Nrf2 signaling pathway and promoted locomotor recovery of SCI mice, which is suggested as a potential target on SCI pathogenesis and treatment.

Oxidative Stress in Roots: Detection of Lipid Peroxidation and Total Antioxidative Capacity.

Various abiotic and biotic agents disturb the fine balance between cellular oxidants and antioxidants. The resulting oxidative stress occurs either due to the increasing levels of reactive oxygen species (ROS) or weak antioxidative system that cannot scavenge ROS burst. In addition to their harmful role, ROS can also act as signaling molecules, and oxidative stress is often the initial step in the programmed cell death. Here we describe two parameters of oxidative stress that can be measured spectrophotometrically: lipid peroxidation via the content of the by-product malondialdehyde, and the amount of all non-enzymatic antioxidants named as total antioxidative capacity. Both methods are presented using young radish (Raphanus sativus) seedlings after treatment with extract from the invasive plant species Japanese knotweed (Fallopia japonica).

Detection of lipid peroxidation in dog spermatozoa with the fluorescent dye C11-BODIPY581/591.

C11-BODIPY581/591 is a fluorescent probe that has been successfully used to evaluate lipid peroxidation in different species, but it has not been completely studied in the dog. Thus, the aim of the present study was to assess lipid peroxidation of dog spermatozoa using C11-BODIPY581/591 and compare different positive controls of the technique. Twenty-four ejaculates were collected from 8 adult male dogs. Routine seminal characteristics were evaluated in raw semen. Lipid peroxidation evaluation was performed as described in other species. Samples were divided in three aliquots, exposed to UV radiation, incubated with hydrogen peroxide or left without treatment (control). Lipid peroxidation was significantly greater only in UV-exposed samples than in the control ones (91±6% vs. 8.3±3.5%, p˂.01). In conclusion, C11-BODIPY581/591 is useful to evaluate lipid peroxidation of dog spermatozoa and UV radiation is a good promoter of membrane oxidation, so irradiated samples can be used as a positive control of this technique.

Self-assembled Co-doped β-Bi2O3 flower-like structure for enhanced photocatalytic antibacterial effect under visible light

A series of β-Bi2O3 with different Co doping amounts were prepared by a co-precipitation and pyrolysis process. A flower-like self-assembly of β-Bi2O3 sheets was realized by the doping of Co. Simulation results showed that an impurity state was introduced into the band gap of doped β-Bi2O3, mainly due to the Co 3d orbital. Co doping enhanced the visible light utilization rate by reducing the band gap and improving electrons holes separation efficiency. E. coli was used to evaluate the photocatalytic antibacterial ability of undoped and Co-doped β-Bi2O3 under visible light. β-Bi2O3 exhibited optimal efficiency for both electron-hole separation and antibacterial effect when doped with 0.4% Co (0.4%-CBO). Gradient experiments were performed to determine that reactive oxygen species (ROS) were the main factor behind the antibacterial effect of β-Bi2O3. The enhanced antibacterial properties of 0.4%-CBO were due to its increased production of O2–, OH, and H2O2, with scavenger experiments finding that O2– was the main contributor to its antibacterial effect. Through the detection of lipid peroxidation and bacterial respiratory chain dehydrogenase activity, it was determined that the antibacterial effect of ROS simultaneously occurred in the interior and exterior of E. coli and that this activity was enhanced by Co doping.

Preparation and photocatalytic antibacterial mechanism of porous metastable β-Bi2O3 nanosheets

Effective and safe application of antibacterials has always been an important aspect for their usage. High-efficiency photocatalytic technology driven by visible light for antibacterial action constitutes a practical solution for antibacterial agents and will not harm the human body or the environment. While most studies on β-Bi2O3 materials with good photocatalytic properties under visible light are conducted in the field of optoelectronics, their potential and mechanism as photocatalytic antibacterial agents have not yet been fully explored. Herein, we report the performance of sheet-like metastable β-Bi2O3 material with rich oxygen vacancies and high electron-hole separation efficiency in antibacterial processes, as well as a preliminary exploration of its antibacterial mechanism. The results revealed that the antibacterial activity of the product against E. coli greatly improved in comparison with commercially available α-Bi2O3 owing to its excellent structure and optical properties. In addition, gradient experiments and scavenger experiments have confirmed that the main antibacterial effect of β-Bi2O3 originates from reactive oxygen species (ROS), and the superoxide radical, ·O2−, of generated ROS is the key reactive species in the antibacterial process. Through the detection of lipid peroxidation and bacterial respiratory-chain dehydrogenase activity, several pathways were identified for the excellent antibacterial activity of the product.

Nitric oxide protects against ferroptosis by aborting the lipid peroxidation chain reaction

Ferroptosis is a type of iron-dependent necrotic cell death, which is typically triggered by the depletion of intracellular glutathione (GSH), which is associated with increased lipid peroxidation. Nitric oxide (NO) is a highly reactive gaseous radical mediator with anti-oxidation properties that terminates lipid peroxidation reactions. In the current study, we report the anti-ferroptotic action of NOC18, an NO donor that spontaneously releases NO, in cells under various ferroptotic conditions in vitro. Our results indicate that, when mouse hepatoma Hepa 1–6 cells are incubated with NOC18, cell death induced by various ferroptotic stimuli such as cysteine (Cys) starvation, the inhibition of glutathione peroxidase 4 (GPX4) and treatment with tertiary-butyl hydroperoxide (TBHP) is significantly reduced. Treatment with NOC18 failed to improve the decrease in the levels of Cys or GSH and the accumulation of ferrous iron upon ferroptotic stimuli. The fluorescent intensity of C11-BODIPY581/591, a probe that is used to detect lipid peroxidation products, was increased somewhat by treatment with NOC18 under conditions of Cys starvation, and the accumulation of lipid peroxidation end-products, as evidenced by the levels of 4-hydroxynonenal, were effectively suppressed. The pre-incubation of TBHP with NOC7, a short-lived NO donor completely eliminated its ability to trigger ferroptosis. These collective results indicate that NO exerts a cytoprotective action against various ferroptotic stimuli by aborting the lipid peroxidation chain reaction.

The toxicogenic effect of Terminalia phanerophlebia Engl. & Diels leaf extract on oxidative stress parameters in an in vitro Hek293 model

Context: Medicinal plants are a highly sought-after alternative to current pharmaceutical drugs because they can be locally cultivated, inexpensive and possess minimal adverse effects. Given that Terminalia phanerophlebia (TP) possesses many useful properties and plays a role in modulating lethal diseases, the cytotoxic effect should be evaluated before its application for therapeutic use. Aims: To investigate the oxidative effect and molecular mechanisms of TP on human embryonic kidney (HEK293) cells. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and adenosine triphosphate (ATP) assays were used to determine the cell viability whilst the thiobarbituric acid reactive species (TBARS) assay was used to detect lipid peroxidation. Endogenous antioxidants, catalase, superoxide dismutase, glutathione peroxidase, heat shock protein 70 and nuclear factor erythroid 2-related factor 2 (Nrf2), were used as oxidative stress markers and were detected via western blotting. Results: A decrease in cell viability with an IC50 of 1.36 mg/mL and ATP were noted. The concentration of malondialdehyde (MDA) increased significantly (p<0.005). Superoxide dismutase, Nrf2 and heat shock protein concentrations were increased. However, glutathione, glutathione peroxidase and catalase were depleted. Conclusions: The results obtained suggest that Terminalia phanerophlebia extract is toxicogenic and induced oxidative stress in HEK293 cells.

Lipid Peroxidation Assay Using BODIPY-Phenylbutadiene Probes: A Methodological Overview.

The assessment of reactive oxygen species has increasing importance in biomedical sciences, due to their biological role in signaling pathways and induction of cell damage at low and high concentrations, respectively. Detection of lipid peroxidation with sensing probes such as some BODIPY dyes has now wide application in studies using fluorescent microplate readers, flow cytometry, and fluorescence microscopy. Two phenylbutadiene derivatives of BODIPY are commonly used as peroxidation probes, non-oxidized probes and oxidized products giving red and green fluorescence, respectively. Peculiar features of lipoperoxidation and BODIPY dye properties make this assessment a rather complex process, not exempt of doubts and troubles. Color changes and fluorescence fading that are not due to lipid peroxidation must be taken into account to avoid misleading results. As a characteristic feature of lipoperoxidation is the propagation of peroxyl radicals, pitfalls and advantages of a delayed detection by BODIPY probes should be considered.

Lower cadmium accumulation and higher antioxidative capacity in edible parts of Brassica campestris L. seedlings applied with glutathione under cadmium toxicity.

Glutathione (GSH) is involved in not only plant developmental processes but also plantresponses to abiotic stresses. A hydroponic experiment was performed to explore the protective roles of exogenous GSH in mitigating cadmium (Cd) stress in Brassica campestris L. seedlings by analyzing the morphological and physiological parameters. Results showed that Cd caused severe growth inhibition and Cd accumulation. However, application of GSH significantly mitigated toxic symptoms induced by Cd, including the improvement of the photosynthesis-, plant growth-, and root morphology-related parameters in seedlings under Cd stress. These responses were associated with a striking increase in activities of representative antioxidative enzymes and contents of corresponding non-enzymatic antioxidants. In vivo imaging of O2.- and H2O2, and the detection of lipid peroxidation further demonstrated that increased ability by GSH for Brassica campestris L. seedlings to endure Cd stress was consistent with a striking elevation of ratios of reduced to oxidized glutathione (GSH/GSSG) and ascorbic acid to dehydroascorbic acid (AsA/DHA). Additionally, GSH application increased Cd retained in roots, thus significantly decreased its translocation from root to shoot, ultimately decreased Cd accumulation in shoots. Taken together, our results proved evidence for GSH in ameliorating Cd toxicity via reducing Cd accumulation in shoots and increasing oxidation resistance. Accordingly, application of GSH could be a high-efficiency and promising strategy to decrease Cd concentration in edible parts of Brassica campestris L. in agricultural production.

Metabolic switch induced by Cimicifuga racemosa extract prevents mitochondrial damage and oxidative cell death

BackgroundCimicifuga racemosa extract is a well-established therapy for menopausal symptoms. The mechanisms underlying the multiple therapeutic effects of Cimicifuga extract, e.g. reducing hot flushes and profuse sweating are not well defined. Recent studies revealed pronounced effects of Ze 450, a Cimicifuga racemosa extract that was produced by a standardized procedure, on energy metabolism through activation of AMP-activated protein kinase in vitro and beneficial anti-diabetic effects in vivo. PurposeThe aim of the study was to investigate the effects of Ze 450 on energy metabolism. Since mitochondria are the key regulators of cellular energy homeostasis, we wanted to elucidate whether Ze 450 affects mitochondrial resilience and can provide protection against oxidative damage in neuronal and liver cells. Methods/study designIn this study, we investigated the effects of Ze 450 (1-200 µg/ml) on mitochondrial integrity and function, and cell viability in models of oxidative stress induced by erastin and RSL-3 in neuronal and liver cells. The effects of Ze 450 in control conditions and after induction of oxidative stress were analyzed using FACS for detecting lipid peroxidation (BODIPY), mitochondrial ROS formation (MitoSOX), mitochondrial membrane potential (TMRE) and cell death (AnnexinV/PI staining). Furthermore, we determined metabolic activity (MTT assay), ATP levels and mitochondrial respiration and glycolysis (oxygen consumption rates, extracellular acidification rates; Seahorse). ResultsZe 450 preserved mitochondrial integrity and ATP levels, and prevented mitochondrial ROS formation, loss of mitochondrial membrane potential and cell death. Notably, Cimicifuga racemosa extract alone did not alter mitochondrial ROS levels, and subtle inhibitory effects on cell proliferation were reversed after withdrawal of the extract. In addition, Ze 450 did not exert toxic effects to liver cells, but rather protected these from the oxidative challenge. Further analysis of the mitochondrial oxygen consumption rate and the extracellular acidification rate revealed that Ze 450 mediated a switch from mitochondrial respiration to glycolysis, and this metabolic shift was a prerequisite for the protective effects against oxidative damage. ConclusionIn conclusion, the bioenergetic shift induced by Ze 450 exerted protective effects in different cell types, and offers promising therapeutic potential in age related diseases involving oxidative stress and mitochondrial damage.

Palmitic Acid Curcumin Ester Facilitates Protection of Neuroblastoma against Oligomeric Aβ40 Insult

Background/Aims: The generation of reactive oxygen species (ROS) caused by amyloid-β (Aβ) is considered to be one of mechanisms underlying the development of Alzheimer’s disease. Curcumin can attenuate Aβ-induced neurotoxicity through ROS scavenging, but the protective effect of intracellular curcumin on neurocyte membranes against extracellular Aβ may be compromised. To address this issue, we synthesized a palmitic acid curcumin ester (P-curcumin) which can be cultivated on the cell membrane and investigated the neuroprotective effect of P-curcumin and its interaction with Aβ. Methods: P-curcumin was prepared through chemical synthesis. Its structure was determined via nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). An MTT assay was used to assess Aβ cytotoxicity and the protective effect of P-curcumin on SH-SY5Y cells. The effect of P-curcumin on Aβ-induced ROS production in vitro and in vivo were assessed based on changes in dichlorofluorescein (DCF) fluorescence. A spectrophotometric method was employed to detect lipid peroxidation. To mimic the interaction of P-curcumin on cell membranes with Aβ, liposomes were prepared by thin film method. Finally, the interactions between free P-curcumin and P-curcumin cultivated on liposomes and Aβ were determined via spectrophotometry. Results: A novel derivative, palmitic acid curcumin ester was prepared and characterized. This curcumin, cultivated on the membranes of neurocytes, may prevent Aβ-mediated ROS production and may inhibit the direct interaction between Aβ and the cellular membrane. Furthermore, P-curcumin could scavenge Aβ-mediated ROS as curcumin in vitro and in vivo, and had the potential to prevent lipid peroxidation. Morphological analyses showed that P-curcumin was better than curcumin at protecting cell shape. To examine P-curcumin’s ability to attenuate direct interaction between Aβ and cell membranes, the binding affinity of Aβ to curcumin and P-curcumin was determined. The association constants for free P-curcumin and curcumin were 7.66 × 10⁴ M^-1 and 7.61 × 10⁵ M^-1, respectively. In the liposome-trapped state, the association constants were 3.71 × 10⁵ M^-1 for P-curcumin and 1.44× 10⁶ M^-1 for curcumin. With this data, the thermodynamic constants of P-curcumin association with soluble Aβ (ΔH, ΔS, and ΔG) were also determined. Conclusion: Cultivated curcumin weakened the direct interaction between Aβ and cell membranes and showed greater neuroprotective effects against Aβ insult than free curcumin.

Oxidative stress equilibrium during obstetric event in normal pregnancy

Objective: The aim of this study was to determine malondialdehyde (MDA) concentration as an oxidative stress marker and total antioxidant capacity (TAC) in pregnancy before and after perinatal event.Methods: This study was performed on 200 healthy full-term pregnant women admitted to pregnancy resolution in Maternal-Child Hospital of Durango, Mexico. Oxidative stress and TAC were assessed through detection of lipid peroxidation by quantitation of thiobarbituric acid-reactive substances (TBARS) and TAC through ferric reducing ability of the plasma (FRAP).Results: Our results showed increased levels of MDA after vaginal delivery (VD). TAC was also increased after obstetric event, but it did not differ between VD and caesarean section.Conclusions: We demonstrated that MDA concentrations are increased two hours after obstetric event, and this increase correlates with VD. The TAC was increased as a compensatory mechanism during obstetric event. Another important finding is that women receiving analgesia administration in VD, as well as dexamethasone administration in caesarean section, experienced a protector effect that decreased MDA levels.

Oral administration of fumonisin B1 and T-2 individually and in combination affects hepatic total and mitochondrial membrane lipid profile of rabbits.

Weaned rabbits were fed diets contaminated with 2mg/kg diet T-2 toxin alone, or 10mg/kg diet fumonisin B1 (FB1) alone, and both toxins in combination (2 + 10mg/kg, respectively) compared to a toxin-free control diet. Samplings were performed after 4 weeks (blood and liver). Bodyweight of T-2-fed group was lower after 4 weeks; the liver weight was increased dramatically (threefold of control). Liver total phospholipids (PLs) provided slight alterations in the fatty acid (FA) composition; all three toxin-treated groups showed a decrease in palmitoleic acid (C16:1 n7) proportion. In the liver mitochondrial PL FA composition, margaric acid (C17:0) proportion decreased in the separated toxin treatments compared to the combined setting. Oleic acid (C18:1 n9) proportion was increased and arachidonic acid (C20:4 n6) was decreased in the FB1-treated group, while docosapentaenoic acid (C22:5 n3) was decreased in the separated treatments. The total monounsaturation was significantly higher in the FB1 group's mitochondrial PL FA profile. After 4 weeks, all toxin treatments decreased the blood plasma reduced glutathione and glutathione peroxidase activity, and FB1 increased the plasma sphinganine/sphingosine ratio. Both mycotoxins seem to cross the hepatocellular and the hepatic mitochondrial membrane, without drastic membrane disruption, as assessed from the PL FA composition, but inducing detectable lipid peroxidation.