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Blood Advances Jul 2023Transcription factor Forkhead box P1 (FOXP1) belongs to the same protein family as the FOXOs that are well-known regulators of murine hematopoietic stem progenitor cell...
Transcription factor Forkhead box P1 (FOXP1) belongs to the same protein family as the FOXOs that are well-known regulators of murine hematopoietic stem progenitor cell (HSPC) maintenance via dampening oxidative stress. FOXP1 and FOXOs can play opposite, or similar, roles depending on cell context; they can crossregulate each other's expression. In a previous study, we have shown that FOXP1 contributes to healthy human HSPC and acute myeloid leukemia (AML) cell growth. Here, we investigated the role of FOXP1 in HSPCs and AML cell oxidative stress defense in a human context. FOXP1 expression level was associated with an inferior survival outcome in patients with cytogenetically normal AML. FOXP1 knockdown enhanced superoxide anion levels of human-committed CD34+CD38+ cells but not stem cell-enriched CD34+CD38- HSPCs or AML cells in vitro. FOXP1 knockdown triggered enhanced NRF2 activity and increased cell oxidative stress. FOXP1 had no impact on FOXO1/3/4 expression in these cells; genetic and pharmacological inhibition of FOXOs did not change superoxide anion levels of human HSPCs or AML cells. Moreover, FOXP1 antioxidant activity was independent of changes in expression of superoxide dismutase 1 and 2 or catalase. Instead, FOXP1 upregulated expression of the stress sensor SIRT1 by stabilizing SIRT1 protein. FOXP1 loss sensitized AML cells to chemotherapy. Together, this study identified FOXP1 as a new safeguard against myeloid progenitor oxidative stress, which works independently of FOXOs but through SIRT1 and contributes to AML chemoresistance. It proposes FOXP1 expression/activity as a promising target to overcome drug resistance of AML HSPCs.
Topics: Humans; Animals; Mice; Sirtuin 1; Superoxides; Leukemia, Myeloid, Acute; Hematopoietic Stem Cells; Oxidative Stress; Repressor Proteins; Forkhead Transcription Factors
PubMed: 36930820
DOI: 10.1182/bloodadvances.2022008585 -
International Immunopharmacology Mar 2024Gestational diabetes mellitus (GDM) is associated with adverse myocardial remodeling and impaired cardiac function of fetus. Nevertheless, specific molecular mechanisms...
BACKGROUND
Gestational diabetes mellitus (GDM) is associated with adverse myocardial remodeling and impaired cardiac function of fetus. Nevertheless, specific molecular mechanisms underlying type 1 GDM-induced fetal myocardial injury remain unknown. Therefore, this study proposes to identify possible molecular mechanisms using RNA-seq.
METHODS
A rat type 1 GDM model was developed using streptozotocin (STZ) (25 and 50 mg/kg), and weight and glucose tolerance of maternal and offspring were evaluated. Changes in markers of myocardial injury and oxidative stress identified by ELISA and biochemical kits in offspring hearts. Identification of differentially expressed mRNAs (DE-mRNAs) associated with myocardial injury in type 1 GDM offspring using RNA-seq. Proliferation, apoptosis, and oxidative stress were assessed in high glucose-induced H9C2 cells after exogenously modulating ATP Synthase Membrane Subunit E (ATP5me).
RESULTS
Maternal weight, glucose and glucose tolerance, and fetal weight and heart weight were reduced in the type 1 GDM model, especially in 50 mg/kg STZ-induced. Increased of creatine kinase-MB (CK-MB), cardiac troponin T (cTnT), hypersensitive C-reactive protein (hs-CRP), reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased of superoxide dismutase (SOD) were observed in type 1 GDM offspring hearts. type 1 GDM offspring hearts exhibited disorganized cardiomyocytes with enlarged gaps, broken myocardial fibers, erythrocyte accumulation and inflammatory infiltration. RNA-seq identified 462 DE-mRNAs in type 1 GDM offspring hearts, which mainly regulate immunity, redox reactions, and cellular communication. Atp5me was under-expressed in type 1 GDM offspring hearts, and high glucose decreased Atp5me expression in H9C2 cells. Overexpressing Atp5me alleviated high glucose-induced decrease in proliferation, mitochondrial membrane potential, BCL2 and SOD, and increase in apoptosis, MDA, ROS, c-Caspase-3, and BAX in H9C2 cells.
CONCLUSION
This study first demonstrated that ATP5me attenuated type 1 GDM-induced fetal myocardial injury. This provides a possible molecular mechanism for the treatment of type 1 GDM-induced fetal myocardial injury.
Topics: Rats; Animals; Myocytes, Cardiac; Reactive Oxygen Species; Myocardium; Oxidative Stress; Glucose; Apoptosis; Heart Injuries; Superoxide Dismutase
PubMed: 38320353
DOI: 10.1016/j.intimp.2024.111626 -
Redox Biology Aug 2023Zinc (Zn) has antioxidant, anti-inflammatory and anti-proliferative actions, with Zn dysregulation associated with coronary ischemia/reperfusion injury and smooth muscle...
Zinc (Zn) has antioxidant, anti-inflammatory and anti-proliferative actions, with Zn dysregulation associated with coronary ischemia/reperfusion injury and smooth muscle cell dysfunction. As the majority of studies concerning Zn have been conducted under non-physiological hyperoxic conditions, we compare the effects of Zn chelation or supplementation on total intracellular Zn content, antioxidant NRF2 targeted gene transcription and hypoxia/reoxygenation-induced reactive oxygen species generation in human coronary artery smooth muscle cells (HCASMC) pre-adapted to hyperoxia (18 kPa O) or normoxia (5 kPa O). Expression of the smooth muscle marker SM22-α was unaffected by lowering pericellular O, whereas calponin-1 was significantly upregulated in cells under 5 kPa O, indicating a more physiological contractile phenotype under 5 kPa O. Inductively coupled plasma mass spectrometry established that Zn supplementation (10 μM ZnCl + 0.5 μM pyrithione) significantly increased total Zn content in HCASMC under 18 but not 5 kPa O. Zn supplementation increased metallothionein mRNA expression and NRF2 nuclear accumulation in cells under 18 or 5 kPa O. Notably, NRF2 regulated HO-1 and NQO1 mRNA expression in response to Zn supplementation was only upregulated in cells under 18 but not 5 kPa. Furthermore, whilst hypoxia increased intracellular glutathione (GSH) in cells pre-adapted to 18 but not 5 kPa O, reoxygenation had negligible effects on GSH or total Zn content. Reoxygenation-induced superoxide generation in cells under 18 kPa O was abrogated by PEG-superoxide dismutase but not by PEG-catalase, and Zn supplementation, but not Zn chelation, attenuated reoxygenation-induced superoxide generation in cells under 18 but not 5kPaO, consistent with a lower redox stress under physiological normoxia. Our findings highlight that culture of HCASMC under physiological normoxia recapitulates an in vivo contractile phenotype and that effects of Zn on NRF2 signaling are altered by oxygen tension.
Topics: Humans; Coronary Vessels; NF-E2-Related Factor 2; Antioxidants; Superoxides; Zinc; Hypoxia; Myocytes, Smooth Muscle; Hyperoxia; Glutathione; RNA, Messenger; Dietary Supplements
PubMed: 37315344
DOI: 10.1016/j.redox.2023.102777 -
Open Veterinary Journal Dec 2023We previously reported that myocardial fibrosis may be one of the causes of left ventricular hypertrophy and cardiac dysfunction in dogs with hyperglucocorticism (HGC)....
BACKGROUND
We previously reported that myocardial fibrosis may be one of the causes of left ventricular hypertrophy and cardiac dysfunction in dogs with hyperglucocorticism (HGC). The detailed mechanism by which myocardial fibrosis of the left ventricle occurs in dogs with HGC remains unclear.
AIM
Th is study investigated the mechanism by which HGC causes fibrosis of the left ventricle.
METHODS
The impa cts of HGC on the heart by comparing samples obtained from high-dose glucocorticoid (GC)-treated (P) and untreated (C) dogs. The P group included healthy Beagle dogs ( = 6) treated with prednisolone (2 mg/kg, bid, po) for 84 days, and the C group included healthy Beagle dogs ( = 6) euthanized for unrelated reasons. In three of the P group dogs, serum was collected before the start of administration (Day 0) and on Day 84 to measure angiotensin II concentrations and oxidative stress markers (8-hydroxy-2'-deoxyguanosine (8OHdG), NADPH oxidase, and superoxide levels). Samples of the left ventricular free wall (LVFW), right ventricular free wall (RVFW), interventricular septum (IVS), and aortic root were harvested from both groups ( = 6 for each group). Using these tissue samples, angiotensin II type 1 receptor (AT1R), 8OHdG, and transforming growth factor β1 (TGFβ1) immunohistochemical stains were performed.
RESULTS
The blood N ADPH oxidase concentration was significantly higher ( = 0.027) in the P group 84 days after initiation of the medication compared to that before prednisolone treatment. By contrast, there was no significant difference in serum angiotensin II ( = 0.450), 8OHdG ( = 0.068), and superoxide ( = 0.057) concentrations. The positive staining rates of AT1R, 8OHdG, and TGFβ1 in the heart (LVFW, RVFW, IVS, and aortic root) were significantly higher in the P group than those in the C group.
CONCLUSION
Angiotensin II and oxidative stress in HGC may cause left ventricular fibrosis in dogs.
Topics: Animals; Dogs; Angiotensin II; Prednisolone; 8-Hydroxy-2'-Deoxyguanosine; Superoxides; Fibrosis; Dog Diseases
PubMed: 38292726
DOI: 10.5455/OVJ.2023.v13.i12.19 -
International Journal of Molecular... Oct 2023The interaction of the activating transcription factor 6 (ATF6), a key effector of the unfolded protein response (UPR) in the endoplasmic reticulum, with the neuronal...
The interaction of the activating transcription factor 6 (ATF6), a key effector of the unfolded protein response (UPR) in the endoplasmic reticulum, with the neuronal calcium sensor Downstream Regulatory Element Antagonist Modulator (DREAM) is a potential therapeutic target in neurodegeneration. Modulation of the ATF6-DREAM interaction with repaglinide (RP) induced neuroprotection in a model of Huntington's disease. Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder with no cure, characterized by the progressive loss of motoneurons resulting in muscle denervation, atrophy, paralysis, and death. The aim of this work was to investigate the potential therapeutic significance of DREAM as a target for intervention in ALS. We found that the expression of the DREAM protein was reduced in the spinal cord of SOD1G93A mice compared to wild-type littermates. RP treatment improved motor strength and reduced the expression of the ALS progression marker collagen type XIXα1 ( mRNA) in the quadriceps muscle in SOD1G93A mice. Moreover, treated SOD1G93A mice showed reduced motoneuron loss and glial activation and increased ATF6 processing in the spinal cord. These results indicate that the modulation of the DREAM-ATF6 interaction ameliorates ALS symptoms in SOD1G93A mice.
Topics: Mice; Animals; Mice, Transgenic; Amyotrophic Lateral Sclerosis; Activating Transcription Factor 6; Neuroprotection; Motor Neurons; Kv Channel-Interacting Proteins; Superoxide Dismutase; Disease Models, Animal
PubMed: 37958767
DOI: 10.3390/ijms242115783 -
PloS One 2024Superoxide dismutase (SOD) is an antioxidant enzyme that protects the body from free radicals. It has both antioxidant and immunomodulatory properties, inducing...
Superoxide dismutase (SOD) is an antioxidant enzyme that protects the body from free radicals. It has both antioxidant and immunomodulatory properties, inducing macrophage polarization from M1 to M2. Macrophages, key mediators of the innate immune response, are divided into the M1 (pro-inflammatory) and M2 (anti-inflammatory) subtypes. In this study, we aimed to assess the antioxidant and neuroprotective effects of SOD on nerve cells and its immunomodulatory effects on macrophages. We observed that SOD inhibited the accumulation of reactive oxygen species and enhanced the viability of H2O2-treated nerve cells. Furthermore, SOD reduced the degree of necrosis in nerve cells treated with the conditioned medium from macrophages, which induced inflammation. In addition, SOD promoted the M1 to M2 transition of macrophages. Our findings suggest that SOD protects nerve cells and regulates immune responses.
Topics: Animals; Superoxide Dismutase; Mice; Macrophages; Humans; Neuroprotective Agents; RAW 264.7 Cells; Reactive Oxygen Species; Neuroblastoma; Cell Line, Tumor; Hydrogen Peroxide; Cell Survival; Antioxidants
PubMed: 38743689
DOI: 10.1371/journal.pone.0303136 -
Muscle & Nerve Oct 2023Electromyography (EMG) remains a key component of the diagnostic work-up for suspected neuromuscular disease, but it does not provide insight into the molecular...
INTRODUCTION/AIMS
Electromyography (EMG) remains a key component of the diagnostic work-up for suspected neuromuscular disease, but it does not provide insight into the molecular composition of muscle which can provide diagnostic information. Raman spectroscopy is an emerging neuromuscular biomarker capable of generating highly specific, molecular fingerprints of tissue. Here, we present "optical EMG," a combination of EMG and Raman spectroscopy, achieved using a single needle.
METHODS
An optical EMG needle was created to collect electrophysiological and Raman spectroscopic data during a single insertion. We tested functionality with in vivo recordings in the SOD1 mouse model of amyotrophic lateral sclerosis (ALS), using both transgenic (n = 10) and non-transgenic (NTg, n = 7) mice. Under anesthesia, compound muscle action potentials (CMAPs), spontaneous EMG activity and Raman spectra were recorded from both gastrocnemius muscles with the optical EMG needle. Standard concentric EMG needle recordings were also undertaken. Electrophysiological data were analyzed with standard univariate statistics, Raman data with both univariate and multivariate analyses.
RESULTS
A significant difference in CMAP amplitude was observed between SOD1 and NTg mice with optical EMG and standard concentric needles (p = .015 and p = .011, respectively). Spontaneous EMG activity (positive sharp waves) was detected in transgenic SOD1 mice only. Raman spectra demonstrated peaks associated with key muscle components. Significant differences in molecular composition between SOD1 and NTg muscle were identified through the Raman spectra.
DISCUSSION
Optical EMG can provide standard electrophysiological data and molecular Raman data during a single needle insertion and represents a potential biomarker for neuromuscular disease.
Topics: Mice; Animals; Electromyography; Superoxide Dismutase-1; Spectrum Analysis, Raman; Muscle, Skeletal; Mice, Transgenic; Amyotrophic Lateral Sclerosis; Disease Models, Animal; Superoxide Dismutase
PubMed: 37477391
DOI: 10.1002/mus.27937 -
Acta Cirurgica Brasileira 2023To investigate the role of puerarin on renal fibrosis and the underlying mechanism in renal ischemia and reperfusion (I/R) model.
PURPOSE
To investigate the role of puerarin on renal fibrosis and the underlying mechanism in renal ischemia and reperfusion (I/R) model.
METHODS
Rats were intraperitoneally injected with puerarin (50 or 100 mg/kg) per day for one week before renal I/R. The level of renal collagen deposition and interstitial fibrosis were observed by hematoxylin and eosin and Sirius Red staining, and the expression of α-smooth muscle actin (α-SMA) was examined by immunohistochemical staining. The ferroptosis related factors and TLR4/Nox4-pathway-associated proteins were detected by Western blotting.
RESULTS
Puerarin was observed to alleviate renal collagen deposition, interstitial fibrosis and the α-SMA expression induced by I/R. Superoxide dismutase (SOD) activities and glutathione (GSH) level were decreased in I/R and hypoxia/reoxygenation (H/R), whereas malondialdehyde (MDA) and Fe2+ level increased. However, puerarin reversed SOD, MDA, GSH and Fe2+ level changes induced by I/R and H/R. Besides, Western blot indicated that puerarin inhibited the expression of ferroptosis related factors in a dose-dependent manner, which further demonstrated that puerarin had the effect to attenuate ferroptosis. Moreover, the increased expression of TLR/Nox4-pathway-associated proteins were observed in I/R and H/R group, but puerarin alleviated the elevated TLR/Nox4 expression.
CONCLUSIONS
Our results suggested that puerarin inhibited oxidative stress and ferroptosis induced by I/R and, thus, delayed the progression of renal fibrosis, providing a new target for the treatment of renal fibrosis.
Topics: Rats; Animals; Ferroptosis; Toll-Like Receptor 4; Oxidative Stress; Reperfusion Injury; Kidney Diseases; Ischemia; Fibrosis; Superoxide Dismutase; NADPH Oxidase 4
PubMed: 37556718
DOI: 10.1590/acb382523 -
Scientific Reports Aug 2023Castanopsis fissa is a native, broadleaf tree species in Guangdong with characteristics of barrenness and fast growth and is often used as a pioneer species for...
Castanopsis fissa is a native, broadleaf tree species in Guangdong with characteristics of barrenness and fast growth and is often used as a pioneer species for vegetation restoration with excellent ecological benefits. To explore the response of C.fissa to drought, this study investigated the drought tolerance mechanism of C.fissa using physiological and proteomic assessments. Using a potted continuous drought experimental method with normal water supply as a control, we measured photosynthetic parameters, antioxidant enzyme activities, and osmoregulatory substances of C. fissa in response to drought stress for 1 to 4 weeks, respectively. In addition, we used TMT quantitative proteomics to identify differentially expressed proteins (DEPs) between the drought-stress-treated C. fissa leaves and the control leaves. With the extension of drought stress time, the photosynthetic indexes and peroxidase (POD) activity of C. fissa leaves showed a decreasing trend. The malondialdehyde (MDA) content; superoxide Dismutase (SOD) and catalase (CAT) activities; and proline (Pro), soluble sugar (SS) and soluble protein (SP) contents showed an overall increasing trend, all of which reached significant differences at 4 w of stress. We identified 177 and 529 DEPs in the 2 and 4 weeks drought-stress leaves, respectively, in reference to the control leaves. These DEPs were closely related to physiological metabolic processes such as photosynthesis, energy and carbohydrate metabolism, stress response and defense, transcriptional regulation, and signal ion transduction. Drought stress mainly affects photosynthesis, carbohydrate metabolism, and protein synthesis and degradation in C. fissa leaves. At 2 weeks of stress, the expression of carbon metabolism, pyruvate metabolism and ribosome-related proteins was significantly changed, however, and at 4 weeks of stress, protein processing in the endoplasmic reticulum and spliceosome-related proteins were significantly increased in plant leaves. To alleviate the effect of water unavailability, the drought-stressed C.fissa leaves increased its oxidative protective enzyme system to eliminate excess reactive oxygen species (ROS) and also increased its Pro and SP contents to maintain the intracellular osmotic potential balance.
Topics: Droughts; Proteomics; Antioxidants; Reactive Oxygen Species; Photosynthesis; Superoxide Dismutase; Stress, Physiological; Plant Leaves
PubMed: 37532761
DOI: 10.1038/s41598-023-39235-x -
Antioxidants (Basel, Switzerland) Sep 2023Amyotrophic Lateral Sclerosis (ALS) is a progressive motor neurodegenerative disease. Cell damage in ALS is the result of many different, largely unknown, pathogenetic... (Review)
Review
Amyotrophic Lateral Sclerosis (ALS) is a progressive motor neurodegenerative disease. Cell damage in ALS is the result of many different, largely unknown, pathogenetic mechanisms. Astrocytes and microglial cells play a critical role also for their ability to enhance a deranged inflammatory response. Excitotoxicity, due to excessive glutamate levels and increased intracellular Ca concentration, has also been proposed to play a key role in ALS pathogenesis/progression. Reactive Oxygen Species (ROS) behave as key second messengers for multiple receptor/ligand interactions. ROS-dependent regulatory networks are usually mediated by peroxides. Superoxide Dismutase 1 (SOD1) physiologically mediates intracellular peroxide generation. About 10% of ALS subjects show a familial disease associated with different gain-of-function SOD1 mutations. The occurrence of sporadic ALS, not clearly associated with SOD1 defects, has been also described. SOD1-dependent pathways have been involved in neuron functional network as well as in immune-response regulation. Both, neuron depolarization and antigen-dependent T-cell activation mediate SOD1 exocytosis, inducing increased interaction of the enzyme with a complex molecular network involved in the regulation of neuron functional activity and immune response. Here, alteration of SOD1-dependent pathways mediating increased intracellular Ca levels, altered mitochondria functions and defective inflammatory process regulation have been proposed to be relevant for ALS pathogenesis/progression.
PubMed: 37760050
DOI: 10.3390/antiox12091747