ROS Overproduction Research Articles

Histone deacetylase inhibitor panobinostat in combination with rapamycin confers enhanced efficacy against triple-negative breast cancer

Triple-negative breast cancer (TNBC) accounts for about 15% of diagnosed breast cancer patients, which has a poor survival outcome owing to a lack of effective therapies. This study aimed to explore the in vitro and in vivo efficiency of histone deacetylase (HDAC) inhibitor panobinostat (PANO) in combination with mTOR inhibitor rapamycin (RAPA) against TNBC. TNBC cells were treated with PANO, RAPA alone or the combination of drugs, then cell growth and apoptosis were evaluated by CCK-8, colony formation and flow cytometry. Cell migration and invasion were detected by wound healing assay and transwell assay, respectively. ROS production was detected by DCFH-DA staining. Western blotting was performed to detect protein levels. In vivo tumor growth was assessed in nude mice. The expression of cleaved caspase-3 and Ki-67 in tumor tissues was detected by immunofluorescence staining. H&E staining was conducted to observe the pathological changes in heart, liver, and kidney tissues. The combination of PANO and RAPA exerted a stronger role in repressing growth, migration, invasion, and inducing apoptosis of TNBC cells compared with monotherapy. Furthermore, this combination presented a more effective anti-cancer efficacy than a single treatment in the xenograft model without apparent toxic side effects. Importantly, mechanistic studies indicated that PANO and RAPA combination led to ROS overproduction, which subsequently activated endoplasmic reticulum stress. Conclusion: PANO in combination with RAPA exhibits enhanced efficacy against TNBC, which may be considered a promising therapeutic candidate.

Ameliorative effect of the probiotic peptide against benzo(α)pyrene-induced inflammatory damages in enterocytes

Probiotics are living bacteria that provide health benefits to the host when consumed in sufficient quantities. However, the protective effect of the bioactive peptides isolated from the probiotics against benzo(α)pyrene (BaP) induced gastrointestinal injury has never been investigated. The current work used a bio-assay guided technique to identify-four new cyclic peptides in BaP-induced Caco-2 cell culture and mouse colitis model. Lactobacillus rhamnosus cycle (Thr-His-Ala-Trp) peptide-1 (LRCP-1) effectively inhibited BaP-induced epithelial cytokine over-release and intracellular ROS over-production. Simultaneously, LRCP-1 attenuated BaP-induced NAD (P)H: oxidases (NOXs), Matrix metalloproteinases (MMPs) over-expression, respectively. Furthermore, increased NAD (P)H: quinone oxidoreductase 1 (NQO1)/heme oxygenase-1 (HO-1)/nuclear factor E2-related factor 2 (Nrf2) expression and aryl hydrocarbon receptor (AhR) pathway activation induced by the BaP-exposure were also inhibited after the LRCP-1 treatment. Notably, LRCP-1 is a promising agent protecting gastrointestinal epithelial cells from BaP-induced inflammatory and oxidative damages.

Response pathways of superoxide dismutase and catalase under the regulation of triclocarban-triggered oxidative stress in Eisenia foetida: Comprehensive mechanism analysis based on cytotoxicity and binding model

Triclocarban (TCC) is an emerging environmental contaminant, posing potential ecological risks. Displaying a high accumulation effect and 120-day half-life in the soil environment, the toxic effects of TCC to soil organisms have been widely reported. Previous studies have confirmed that TCC can induce the oxidative stress and changes in superoxide dismutase (SOD) and catalase (CAT) activities in earthworms, but the underlying mechanisms of oxidative stress and disorder in antioxidant enzyme activities induced by TCC have not yet been elucidated. Here, we explored the multiple response mechanisms of SOD and CAT under the regulation of oxidative stress induced by TCC. Results indicated that higher-dose (0–2.0 mg/L) TCC exposure triggered the overproduction of ROS in Eisenia foetida coelomocytes, causing oxidative damage and a decrease in cell viability that was response to ROS accumulation. The TCC-induced inhibition of intracellular SOD/CAT activity was found under the regulation of oxidative stress (SOD: 29.2 %; CAT: 18.5 %), and this effect was blunted by antioxidant melatonin. At the same time, the interaction between antioxidative enzymes and TCC driven by various forces (SOD: electrostatic interactions; CAT: van der Waals forces and hydrogen bonding) led to inhibited SOD activity (9.84 %) and enhanced CAT activity (17.5 %). Then, to elucidate the binding mode of TCC, we explored the changes in SOD and CAT structure (protein backbone and secondary structure), the microenvironment of aromatic amino acids, and aggregation behavior through multispectral techniques. Molecular docking results showed that TCC inhibited SOD activity in a substrate competitive manner and enhanced CAT activity by the stabilizing effects of TCC on the heme groups. Collectively, this study reveals the response mechanisms of SOD/CAT under the regulation of TCC-triggered oxidative stress and shed a new light on revealing the toxic pathways of exogenous pollutants on antioxidant-related proteins function.

Effect of Modified Levopimaric Acid Diene Adducts on Mitochondrial and Liposome Membranes.

This paper demonstrates the membranotropic effect of modified levopimaric acid diene adducts on liver mitochondria and lecithin liposomes. We found that the derivatives dose-dependently reduced the efficiency of oxidative phosphorylation of mitochondria due to inhibition of the activity of complexes III and IV of the respiratory chain and protonophore action. This was accompanied by a decrease in the membrane potential in the case of organelle energization both by glutamate/malate (complex I substrates) and succinate (complex II substrate). Compounds 1 and 2 reduced the generation of H2O2 by mitochondria, while compound 3 exhibited a pronounced antioxidant effect on glutamate/malate-driven respiration and, on the other hand, caused ROS overproduction when organelles are energized with succinate. All tested compounds exhibited surface-active properties, reducing the fluidity of mitochondrial membranes and contributing to nonspecific permeabilization of the lipid bilayer of mitochondrial membranes and swelling of the organelles. Modified levopimaric acid diene adducts also induced nonspecific permeabilization of unilamellar lecithin liposomes, which confirmed their membranotropic properties. We discuss the mechanisms of action of the tested compounds on the mitochondrial OXPHOS system and the state of the lipid bilayer of membranes, as well as the prospects for the use of new modified levopimaric acid diene adducts in medicine.

Engineering cancer cell membrane-camouflaged metal complex for efficient targeting therapy of breast cancer

BackgroundCancer cell membrane-camouflaged nanotechnology for metal complex can enhance its biocompatibility and extend the effective circulation time in body. The ruthenium polypyridyl complex (RuPOP) has extensive antitumor activity, but it still has disadvantages such as poor biocompatibility, lack of targeting, and being easily metabolized by the organism. Cancer cell membranes retain a large number of surface antigens and tumor adhesion molecules CD47, which can be used to camouflage the metal complex and give it tumor homing ability and high biocompatibility.ResultsTherefore, this study provides an electrostatic adsorption method, which uses the electrostatic interaction of positive and negative charges between RuPOP and cell membranes to construct a cancer cell membrane-camouflaged nano-platform (RuPOP@CM). Interestingly, RuPOP@CM maintains the expression of surface antigens and tumor adhesion molecules, which can inhibit the phagocytosis of macrophage, reduce the clearance rate of RuPOP, and increase effective circulation time, thus enhancing the accumulation in tumor sites. Besides, RuPOP@CM can enhance the activity of cellular immune response and promote the production of inflammatory cytokines including TNF-α, IL-12 and IL-6, which is of great significance in treatment of tumor. On the other hand, RuPOP@MCM can produce intracellular ROS overproduction, thereby accelerating the apoptosis and cell cycle arrest of tumor cells to play an excellent antitumor effect in vitro and in vivo.ConclusionIn brief, engineering cancer cell membrane-camouflaged metal complex is a potential strategy to improve its biocompatibility, biological safety and antitumor effects.

Octocrylene exposure impairs mouse oocyte quality by inducing spindle defects and mitochondria dysfunction

One of organic ultraviolet (UV) filters, Octocrylene (OCL), is mainly used in various cosmetic products, which is being frequently detected in soil, sediment, aquatic systems and food chain. There is evidence confirmed the reproductive toxicity of OCL in Japanese medaka. However, less was known about the effects of OCL exposure on oocyte quality. Here, we investigated the impacts of OCL on mouse oocyte maturation and quality by exposing oocytes to OCL in vitro at concentrations of 8, 22, 30, 40 and 50 nM. The results showed that OCL markedly reduced mouse oocyte germinal vesicle breakdown (GVBD) at 50 nM and polar body extrusion (PBE) rates at 40 and 50 nM. OCL exposure further disrupted spindle assembly and chromosome alignment, finally inducing aneuploid. Mitochondrial function was also damaged by OCL exposure, leading to ROS overproduction and apoptosis in oocytes. Moreover, OCL treatment impaired the distribution of cortical granules and sperm binding ability of oocytes. In summary, these data demonstrated that OCL could disturb the oocyte meiotic maturation and reduce oocyte quality.

The Strong Anti-Tumor Effect of Smp24 in Lung Adenocarcinoma A549 Cells Depends on Its Induction of Mitochondrial Dysfunctions and ROS Accumulation.

Non-small cell lung cancer (NSCLC) is the leading cause of death in lung cancer due to its aggressiveness and rapid migration. The potent antitumor effect of Smp24, an antimicrobial peptide derived from Egyptian scorpion Scorpio maurus palmatus via damaging the membrane and cytoskeleton have been reported earlier. However, its effects on mitochondrial functions and ROS accumulation in human lung cancer cells remain unknown. In the current study, we discovered that Smp24 can interact with the cell membrane and be internalized into A549 cells via endocytosis, followed by targeting mitochondria and affect mitochondrial function, which significantly causes ROS overproduction, altering mitochondrial membrane potential and the expression of cell cycle distribution-related proteins, mitochondrial apoptotic pathway, MAPK, as well as PI3K/Akt/mTOR/FAK signaling pathways. In summary, the antitumor effect of Smp24 against A549 cells is related to the induction of apoptosis, autophagy plus cell cycle arrest via mitochondrial dysfunction, and ROS accumulation. Accordingly, our findings shed light on the anticancer mechanism of Smp24, which may contribute to its further development as a potential agent in the treatment of lung cancer cells.

Effect of UV-B Irradiation on Bioactive Compounds of Red Perilla (Perilla frutescens (L.) Britton) Cultivated in a Plant Factory with Artificial Light

In this study, we investigated the effect of UV-B irradiation 3 days prior to harvest, on the accumulation of rosmarinic acid (RA) and anthocyanin, and the expression of genes related to phenylpropanoid and flavonoid biosynthetic pathways, in red perilla (Perilla frutescens L.). In experiment 1, seedlings at 60 days after sowing (DAS) were subjected to UV-B irradiation at 0 (control), 6, and 10 W m−2 under a 16 h light period; while in experiment 2, seedlings at 45 DAS were subjected to UV-B irradiation at 0 (control), 4 W m−2 at continuous irradiation, and 6 W m−2 at 16 h irradiation. UV irradiation of 10 W m−2 for 16 h negatively affected leaf color, while irradiation at 6 W m−2 enhanced RA biosynthesis and antioxidant capacity. Continuous UV-B irradiation of 4 W m−2 increased the RA concentration by 92% compared to the control; however, this effect was smaller than that of UV-B irradiation at 6 W m−2 for 16 h, 141% higher than that of the control and had a lower antioxidant capacity against UV-mediated ROS overproduction during the dark period. Results demonstrate that 6 W m−2 of UV-B irradiation for 16 h is suitable for enhancing the RA concentration and content of red perilla.

Effect of metformin on intact mitochondria from liver and brain: Concept revisited

Metformin is an antihyperglycemic drug which is being examined as a repurposed treatment for cardiovascular disease for individuals without diabetes mellitus. Despite evidence that mitochondrial respiratory complex I is a target of metformin and inhibition of the enzyme is one of the mechanisms of its therapeutic actions, no systematic studies of the metformin effect on intact mitochondria have been reported. In the presented paper, we described the effect of metformin on respiration and ROS release by intact mitochondria from the liver and brain. By comparing the effect of metformin on mitochondria oxidizing different substrates, we found direct inhibition of respiration and stimulation of ROS release when complex I-based respiration is measured (forward electron transfer). Metformin had no effect on respiration rates but inhibited ROS release when mitochondria oxidize succinate or glycerol 3-phosphate in conditions of reverse electron transfer in complex I. In addition, we found that metformin is a weak effector of the active/deactive (A/D) transition of mitochondrial complex I. At high concentrations, metformin increases the rate of spontaneous deactivation of complex I (A→D transition). The results obtained are consistent with the concept of metformin inhibition of complex I and that it can either stimulate or inhibit mitochondrial ROS production depending on the preferential respiratory substrate. This is relevant during the ischemia/reperfusion process, to counteract the ROS overproduction, which is induced by a high level of reverse electron transfer substrates is generated after an ischemic event.

Melatonin in cryopreservation media improves transplantation efficiency of frozen–thawed spermatogonial stem cells into testes of azoospermic mice

BackgroundCryostorage of spermatogonial stem cells (SSCs) is an appropriate procedure for long-term storage of SSCs for fertility preservation. However, it causes damage to cellular structures through overproduction of ROS and oxidative stress. In this study, we examined the protective effect of melatonin as a potent antioxidant in the basic freezing medium to establish an optimal cryopreservation method for SSCs.MethodsSSCs were obtained from the testes of neonatal male mice aged 3–6 days. Then, 100 μM melatonin was added to the basic freezing medium containing DMSO for cryopreservation of SSCs. Viability, apoptosis-related markers (BAX and BCL2), and intracellular ROS generation level were measured in frozen–thawed SSCs before transplantation using the MTT assay, immunocytochemistry, and flow cytometry, respectively. In addition, Western blotting and immunofluorescence were used to evaluate the expression of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins in frozen–thawed SSCs after transplantation into recipient testes.ResultsThe data showed that adding melatonin to the cryopreservation medium markedly increased the viability and reduced intracellular ROS generation and apoptosis (by decreasing BAX and increasing BCL2) in the frozen–thawed SSCs (p < 0.05). The expression levels of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins and resumption of spermatogenesis from frozen–thawed SSCs followed the same pattern after transplantation.ConclusionsThe results of this study revealed that adding melatonin as an antioxidant to the cryopreservation medium containing DMSO could be a promising strategy for cryopreservation of SSCs to maintain fertility in prepubertal male children who suffer from cancer.

Linolenic Acid Plus Ethanol Exacerbates Cell Death in Saccharomyces cerevisiae by Promoting Lipid Peroxidation, Cardiolipin Loss, and Necrosis.

Polyunsaturated fatty acids (PUFA) hypersensitize yeast to oxidative stress. Ethanol accumulation during fermentation is another factor that induces oxidative stress via mitochondrial dysfunction and ROS overproduction. Since this microorganism has raised growing interest as a PUFA factory, we have studied if the combination of PUFA plus ethanol enhances yeast death. Respiration, ROS generation, lipid peroxidation, mitochondrial cardiolipin content, and cell death were assessed in yeast grown in the presence of 10% ethanol (ETOH) or linolenic acid (C18:3), or ethanol plus C18:3 (ETOH+C18:3). Lipid peroxidation and cardiolipin loss were several-fold higher in cells with ETOH+C18:3 than with C18:3. On the contrary, ETOH tended to increase cardiolipin content without inducing changes in lipid peroxidation. This was consistent with a remarkable diminution of cell growth and an exacerbated propidium iodide staining in cells with only ETOH+C18:3. The respiration rate decreased with all the treatments to a similar degree, and this was paralleled with similar increments in ROS between all the treatments. These results indicate that PUFA plus ethanol hypersensitize yeast to necrotic cell death by exacerbating membrane damage and mitochondrial cardiolipin loss, independent of mitochondrial dysfunction and ROS generation. The implications of these observations for some biotechnological applications in yeast and its physiology are discussed.

Effects of Different Types of Chronic Training on Bioenergetic Profile and Reactive Oxygen Species Production in LHCN-M2 Human Myoblast Cells.

Human skeletal muscle contains three different types of fibers, each with a different metabolism. Exercise differently contributes to differentiation and metabolism in human myoblast cells. The aims of the present study were to investigate the effects of different types of chronic training on the human LHCN-M2 myoblast cell bioenergetic profile during differentiation in real time and on the ROS overproduction consequent to H2O2 injury. We demonstrated that exercise differently affects the myoblast bioenergetics: aerobic exercise induced the most efficient glycolytic and oxidative capacity and proton leak reduction compared to untrained or anaerobic trained sera-treated cells. Similarly, ROS overproduction after H2O2 stress was lower in cells treated with differently trained sera compared to untrained sera, indicating a cytoprotective effect of training on the reduction of oxidative stress, and thus the promotion of longevity. In conclusion, for the first time, this study has provided knowledge regarding the modifications induced by different types of chronic training on human myoblast cell bioenergetics during the differentiation process in real time, and on ROS overproduction due to stress, with positive implications in terms of longevity.

A triple threat: Ocean warming, acidification, and rare earth elements exposure triggers a superior antioxidant response and pigment production in the adaptable Ulva rigida

• La and Gd were accumulated in 24 h. • Elimination of La and Gd did not occur in U. rigida . • La and Gd showed different accumulation and elimination patterns in future predicted scenarios. • La and Gd triggered an efficient antioxidant defence response in U. rigida . • REE and climate change exposure requested a superior antioxidant response. Anthropogenic increased atmospheric CO 2 concentrations will lead to a drop of 0.4 units of seawater pH and ocean warming up to 4.8°C by 2100. Contaminant's toxicity is known to increase under a climate change scenario. Rare earth elements (REE) are emerging contaminants, that until now have no regulation regarding maximum concentration and discharge into the environment and have become vital to new technologies such as electric and hybrid-electric vehicle batteries, wind turbine generators and low-energy lighting. Studies of REE, namely Lanthanum (La) and Gadolinium (Gd), bioaccumulation, elimination, and toxicity in a multi-stressor environment (e.g., warming and acidification) are lacking. Hence, we investigated the algae phytoremediation capacity, the ecotoxicological responses and total chlorophyll and carotenoid contents in Ulva rigida during 7 days of co-exposure to La or Gd (15 µg L −1 or 10 µg L −1 , respectively), and warming and acidification. Additionally, we assessed these metals elimination, after a 7-day phase. After one day of experiment La and Gd clearly showed accumulation/adsorption in different patterns, at future conditions. Unlikely for Gd, Warming and Acidification contributed to the lowest La accumulation, and increased elimination. Lanthanum and Gd triggered an adequate activation of the antioxidant defence system, by avoiding lipid damage. Nevertheless, REE exposure in a near-future scenario triggered an overproduction of ROS that requested an enhanced antioxidant response. Additionally, an increase in total chlorophyll and carotenoids could also indicate an unforeseen energy expense, as a response to a multi-stressor environment.

Biphasic effects of typical chlorinated organophosphorus flame retardants on Microcystis aeruginosa

The potential accumulation of chlorinated organophosphorus flame retardants (Cl-OPFRs) in aquatic environments sparked interest in studying the effects of Cl-OPFRs on cyanobacterial blooms. In this work, two common Cl-OPFRs, tris(1,3-dichloro-2-propyl) phosphate (TDCPP) and tris(2-chloroethyl) phosphate (TCEP), induced dose-dependent biphasic effect on bloom-forming M. aeruginosa. The hormetic response to low-dose Cl-OPFRs was associated with the upregulation of the type I NADH dehydrogenase (NDH-1) complex and its mediated cyclic electron transfer (CET) pathway, as reflected by a transient post-illumination increase in chlorophyll fluorescence, the dark reduction of P700+ and the change of NDH-1-related gene expression. The increased CET activity and carotenoid content jointly reduced the intracellular ROS production, facilitating cyanobacterial growth. Conversely, a higher concentration of both Cl-OPFRs induced severe inhibition of growth and photosynthetic oxygen-evolving activity through an imbalance between PSII and PSI. Toxic-dose Cl-OPFRs inhibited state transition and fixed cells into the State I with a higher PSII/PSI ratio, as indicated by chlorophyll fluorescence induction, 77 K fluorescence emission spectra and photosystem stoichiometry. The elevated PSII/PSI ratio created an imbalance between the two photosystems and eventually lead to ROS overproduction, which generate adverse effects on cell growth. This work provides important insights into the hormetic mechanism of Cl-OPFRs on Microcystis aeruginosa and their potential roles in harmful cyanobacteria blooms.

Rab27a-dependent exosomes protect against cerebral ischemic injury by reducing endothelial oxidative stress and apoptosis.

IntroductionMulticellular crosstalk within the brain tissue has been suggested to play a critical role in maintaining cerebral vascular homeostasis. Exosomes (EXs) mediated cell–cell communication, but its role in cerebral ischemic injury is largely unknown. Rab27a is one of the major genes controlling EX release. Here, we explored the role of Rab27a in regulating brain EXs secretion, and the effects of Rab27a‐mediated EXs on ischemia evoked cerebral vascular disruption and brain injury.MethodsCerebral ischemia was induced in Rab27a knockout (Rab27a−/−) and wide type (WT) mice by transient middle cerebral artery occlusion (tMCAO). Differential gene expression analysis was performed in ischemic brain tissue by using mRNA sequencing. EXs isolated from brain tissue of Rab27a−/− and WT mice (EXWT or EXRab27a−/−) were pre‐administrated into tMCAO operated Rab27a−/− mice or oxygen and glucose deprivation (OGD) treated primary brain vascular endothelial cells (ECs).ResultsWe demonstrated that Rab27a expression in the peri‐infarct area of brain was significantly elevated, which was associated with local elevation in EXs secretion. Rab27a deficiency dramatically decreased the level of EXs in brain tissue of normal and tMCAO‐treated mice, and Rab27a−/− mice displayed an increase in infarct volume and NDS, and a decrease in cMVD and CBF following tMCAO. Pre‐infusion of EXWT increased the brain EXs levels in the tMCAO operated Rab27a−/− mice, accompanied with an increase in cMVD and CBF, and a decrease in infarct volume, NDS, ROS production, and apoptosis. The effects of EXRab27a−/− infusion were much diminished although in a dose‐dependent manner. In OGD‐treated ECs, EXRab27a−/− showed less effectivity than EXWT in decreasing ROS overproduction and apoptosis, paralleling with down‐regulated expression of NOX2 and cleaved caspase‐3.ConclusionOur study demonstrates that Rab27a controls brain EXs secretion and functions, contributing to cerebral vascular protection from ischemic insult by preventing oxidative stress and apoptosis via down‐regulating NOX2 and cleaved caspase‐3 expression.

Effective Dose of Moringa Leaf Extract (Moringa oleifera Lamk.) to Descrease Total Cholesterol Levels in Streptozotocin-Induced Male Wistar Rats

Lipid metabolic disorder in hyperlipidemia characterized by an increase in the entire lipid profile in the blood. In 2019, the global prevalence of hyperlipidemia was 64.2%. Many patients with diabetes mellitus developed to hyperlipidemia. Diabetes mellitus characterized by hyperglycemia that caused by insulin production defects or decreased insulin resistance. Hyperglycemia in this study resulted from the injection of a streptozotocin dose of 45mg/kgBB intraperitoneally. Hyperglycemia could induced hyperlipidemia. It is due to increased lipid synthesis resulting in overproduction of HMG-CoA and ROS. Moringa leaves contain active substances such as flavonoids and vitamin C that act as antioxidants. This study was conducted to find out the effective dose of moringa leaf extract to lower the total cholesterol levels of male Wistar rats induced by streptozotocin. The dose of moringa leaf extract was divided into 5 groups, namely 62.5 mg/kgBW, 125 mg/kgBW, 250 mg/kgBW, 500 mg/kgBW, and 1000 mg/kgBW. The analysis of the data in this study was a Pearson correlation test that showed p&lt; 0.05. It demonstrated that administration of moringa leaf extract correlated with decreased cholesterol levels of male Wistar rats. The effective dose of moringa leaf extract was calculated using a linear regression test. The equation obtained from the regression test was y= 0.0739x + 153.59, so the dose of moringa leaf extract that effectively lowers the total cholesterol levels of male Wistar rats was 528.96 mg/kgBW.&#x0D; Keywords: hyperlipidemia, diabetes mellitus, cholesterol, Moringa oleifera Lamk., streptozotocin

N-3 PUFA dietary lipid replacement normalizes muscle mitochondrial function and oxidative stress through enhanced tissue mitophagy and protects from muscle wasting in experimental kidney disease

Introduction and methodsSkeletal muscle mitochondrial dysfunction may cause tissue oxidative stress and consequent catabolism in chronic kidney disease (CKD), contributing to patient mortality. We investigated in 5/6-nephrectomized (Nx) rats the impact of n3-polyunsaturated fatty-acids (n3-PUFA) isocaloric partial dietary replacement on gastrocnemius muscle (Gm) mitochondrial master-regulators, ATP production, ROS generation and related muscle-catabolic derangements. ResultsNx had low Gm mitochondrial nuclear respiratory factor-2 and peroxisome proliferator-activated receptor gamma coactivator-1alpha, low ATP production and higher mitochondrial fission-fusion protein ratio with ROS overproduction. n3-PUFA normalized all mitochondrial derangements and pro-oxidative tissue redox state (oxydized to total glutathione ratio). n3-PUFA also normalized Nx-induced muscle-catabolic proinflammatory cytokines, insulin resistance and low muscle weight. Human uremic serum reproduced mitochondrial derangements in C2C12 myotubes, while n3-PUFA coincubation prevented all effects. n3-PUFA also enhanced muscle mitophagy in-vivo and siRNA-mediated autophagy inhibition selectively blocked n3-PUFA-induced normalization of C2C12 mitochondrial ROS production. ConclusionsIn conclusion, dietary n3-PUFA normalize mitochondrial master-regulators, ATP production and dynamics in experimental CKD. These effects occur directly in muscle cells and they normalize ROS production through enhanced mitophagy. Dietary n3-PUFA mitochondrial effects result in normalized catabolic derangements and protection from muscle wasting, with potential positive impact on patient survival.

Parkin, as a Regulator, Participates in Arsenic Trioxide-Triggered Mitophagy in HeLa Cells.

Parkin is a well-established synergistic mediator of mitophagy in dysfunctional mitochondria. Mitochondria are the main target of arsenic trioxide (ATO) cytotoxicity, and the effect of mitophagy on ATO action remains unclear. In this study, we used stable Parkin-expressing (YFP-Parkin) and Parkin loss-of-function mutant (Parkin C431S) HeLa cell models to ascertain whether Parkin-mediated mitophagy participates in ATO-induced apoptosis/cell death. Our data showed that the overexpression of Parkin significantly sensitized HeLa cells to ATO-initiated proliferation inhibition and apoptosis; however, the mutation of Parkin C431S significantly weakened this Parkin-mediated responsiveness. Our further investigation found that ATO significantly downregulated two fusion proteins (Mfn1/2) and upregulated fission-related protein (Drp1). Autophagy was also activated as evidenced by the formation of autophagic vacuoles and mitophagosomes, increased expression of PINK1, and recruitment of Parkin to impaired mitochondria followed by their degradation, accompanied by the increased transformation of LC3-I to LC3-II, increased expression of Beclin1 and decreased expression of P62 in YFP-Parkin HeLa cells. Enhanced mitochondrial fragmentation and autophagy indicated that mitophagy was activated. Furthermore, during the process of mitophagy, the overproduction of ROS implied that ROS might represent a key factor that initiates mitophagy following Parkin recruitment to mitochondria. In conclusion, our findings indicate that Parkin is critically involved in ATO-triggered mitophagy and functions as a potential antiproliferative target in cancer cells.

Regulation of lipid peroxidation in mitochondria by nitroglycerine

The mechanisms of lipid oxidation under the application of nitroglycerine (NG) were studied in isolated rat heart and liver mitochondria. Dose-dependent formation of diene conjugates (DC), leukotriene C4 (LTC4) and thromboxane B2 (TxB2) was shown. To disclose the mechanisms regulating lipid peroxidation in mitochondria, we studied the effect of NG application on the formation of prooxidants (H2O2 and free Fe2+), as well as xanthine oxidase and mtNOS activity as main sources of ROS and RNS. Based on the correlation dependences, we have found that DC, LTC4, and TxB2 formation was strongly dependent on hydroperoxide production and free divalent iron release in mitochondria. Also, DC formation exhibited the dependence on Ca2+ uptake in mitochondria. No dependence of lipid oxidation on xanthine oxidase activity was found. In heart, but not liver mitochondria, DC, LTC4, and TxB2 exhibited strong dependence on mtNOS activity, but were independent of nitrosothiols formation. This indicated that lipid oxidation was independent of direct protein nitrosylation caused by NG application. No dependence of lipid oxidation on mtNOS activity in liver was found, which agreed with much higher mtNOS activity in heart mitochondria, and suppression of mtNOS activity in liver mitochondria at high doses of NG. So, we came to the conclusion that under NG application ROS overproduction and free Fe2+ release promoted both enzymatic and non-enzymatic lipid oxidation in heart and liver mitochondria. Also, we hypothesized that RNS overproduction due to the elevated mtNOS activity in the heart could largely contribute to lipid peroxidation and promote much faster increase in the formation of lipid oxidation products in heart as compared to liver mitochondria, especially at high doses of NG. Obtained correlation dependences allowed us disclose free iron, hydroperoxide, and mtNOS activity as principal factors affecting lipid peroxidation in mitochondria under NG application.

Polystyrene microplastics-induced cardiotoxicity in chickens via the ROS-driven NF-κB-NLRP3-GSDMD and AMPK-PGC-1α axes

Microplastics (MPs) pollution is getting increasingly prominent, and its dangers have attracted widespread attention. The heart is the central hub of the organism's survival, and the mechanism of MPs-induced heart injury in chickens is unknown. Here, we investigated the effects of 5 μm polystyrene microplastics (PS-MPs) on the heart and primary cardiomyocytes of chickens at varied concentrations. We observed that PS-MPs caused severe pathological damage and ultrastructural changes in heart, induced myocardial pyroptosis, inflammatory cell infiltration and mitochondrial lesions. PS-MPs evoked abnormal antioxidant enzyme content and ROS overproduction. Detailed mechanistic investigation indicated that PS-MPs triggered pyroptosis via NF-κB-NLRP3-GSDMD axis and exacerbated myocardial inflammation (NLRP3, Caspase-1, IL-1β, IL-18, ASC, GSDMD, NF-κB, COX-2, iNOS and IL-6 overexpression). Additionally, PS-MPs induced mitochondrial damage (TFAM, OPA1, MFN1 and MFN2 down-expression, DRP1 and Fis1 overexpression) and energy metabolism disorders (HK2, PKM2, PDHX and LDH up-regulation) by inhibiting AMPK-PGC-1α pathway. Interestingly, NAC alleviated these aberrant manifestations in vitro. We suggested that PS-MPs driven alterations in NF-κB-NLRP3-GSDMD and AMPK-PGC-1α pathways via ROS overload, which in turn triggered oxidative stress, myocardial pyroptosis, inflammation, mitochondrial and energy metabolism dysfunction. This provided theoretical bases for protecting chickens from toxic injury by MPs.