Antioxidant Proteins Research Articles

Nrf2 and autonomic dysregulation in chronic heart failure and hypertension.

Redox imbalance plays essential role in the pathogenesis of cardiovascular diseases. Chronic heart failure (CHF) and hypertension are associated with central oxidative stress, which is partly mediated by the downregulation of antioxidant enzymes in the central autonomic neurons that regulate sympathetic outflow, resulting in sympathoexcitation. Antioxidant proteins are partially regulated by the transcriptional factor nuclear factor erythroid 2-related factor 2 (Nrf2). Downregulation of Nrf2 is key to disrupting central redox homeostasis and mediating sympathetic nerve activity in the setting of Chronic heart failure and hypertension. Nrf2, in turn, is regulated by various mechanisms, such as extracellular vesicle-enriched microRNAs derived from several cell types, including heart and skeletal muscle. In this review, we discuss the role of Nrf2 in regulating oxidative stress in the brain and its impact on sympathoexcitation in Chronic heart failure and hypertension. Importantly, we also discuss interorgan communication via extracellular vesicle pathways that mediate central redox imbalance through Nrf2 signaling.

Assessment of Chemical Constituents of <i>Allium sativum</i> Essential Oil Extracted by using Hydrodistillation Technique and their Pharmacological Potential

The current study was based on identification of chemical components and pharmacological potential of Allium sativum essential oil. The pharmacological properties mainly antioxidant, anti-inflammatory and anti-diabetic potential of Allium sativum Essential Oil (ASEO) were also studied. Allium sativum is a traditional medicinal plant of the Alliaceae family. The extracted essential oil from fresh bulbs of Allium sativum through hydro distillation process was further analyzed by GC-MS. To determine the potential of Allium sativum essential oil for reducing inflammation and preventing diabetes, evaluation of antioxidant assays, protein denaturation and glucose absorption by yeast cells were studied. The results explained the presence of several organosulfur compounds that were identified by using different combinations of non-polar solvents mainly represented as Diallyldisulfide, Methyl Allyl Disulfide, Methyl Allyl Trisulfide, Diallyltrisulfide and Diallyltetrasulfide. The antioxidant potential of ASEO expressed as IC50 values of DPPH, ABTS, β-carotene assay, Nitric Oxide, superoxide, hydroxyl scavenging and metal chelating were 5.93 mg/ml, 2.1 mg/ml, 7.94 mg/ml, 5.49 μg/ml, 11.55 μg/ml, 7.66 μg/ml and 10.60 μg/ ml respectively. Further, total phenolic level, total flavonoid level, FRAP assay and total antioxidant capacity were evaluated and they were as follows: 4.99 ± 0.41 mg GAE/g, 98.46 ± 0.16 mg QE/g, 4.99 ± 0.26 mg/g and 15.80 ± 0.14 mg/g. Furthermore, the anti-inflammatory assay was assessed by considering the protein denaturation method which exhibits IC50 value 3.53μg/ml. In addition to this, anti-diabetic potential was assessed by considering glucose absorption by yeast cells at various glucose concentrations (5mM, 10mM, 25mM) which indicated that the ASEO enhanced glucose uptake via the yeast cell membrane. Upregulation of glucose was reported with sequential elevations in the ASEO concentration. Based on the above results, it was revealed that Allium sativum essential oil exhibits reasonable antioxidant, anti-inflammatory and anti-diabetic activity that could be considered as an alternative therapeutic agent in inflammatory diseases. However, future research is required to isolate prominent functional components to determine the observed potential in preclinical studies.

Protective Effect of Daidzein against Diethylnitrosamine/Carbon Tetrachloride-Induced Hepatocellular Carcinoma in Male Rats.

Hepatocellular carcinoma (HCC) is the second-largest cause of death among all cancer types. Many drugs have been used to treat the disease for a long time but have been mostly discontinued because of their side effects or the development of resistance in the patients with HCC. The administration of DZ orally is a great focus to address the clinical crisis. Daidzein (DZ) is a prominent isoflavone polyphenolic chemical found in soybeans and other leguminous plants. It has various pharmacological effects, including anti-inflammatory, antihemolytic, and antioxidant. This present study investigates the protective effect of DZ on chemically induced HCC in rat models. The DZ was administered orally four weeks before HCC induction and continued during treatment. Our study included four treatment groups: control (group 1, without any treatment), HCC-induced rats (group II), an HCC group treated with DZ at 20 mg/kg (group III), and an HCC group treated with DZ at 40 mg/kg (group IV). HCC rats showed elevation in all the HCC markers (AFP, GPC3, and VEGF), liver function markers (ALP, ALT, and AST), inflammatory markers (IL-6, TNF-α, and CRP), and lipid markers concomitant with a decrease in antioxidant enzymes and protein. However, groups III and IV demonstrated dose-dependent alleviation in the previous parameters resulting from HCC. In addition, the high dose of DZ reduces many hepatological changes in HCC rats. All study parameters improved with DZ administration. Due to its antioxidant and anti-inflammatory characteristics, DZ is a promising HCC treatment option for clinical use.

6-Shogaol prevents benzo (A) pyrene-exposed lung carcinogenesis via modulating PRDX1-associated oxidative stress, inflammation, and proliferation in mouse models.

In this study, we have investigated the chemopreventive role of 6-shogaol (6-SGL) on benzopyrene (BaP) exposed lung carcinogenesis by modulating PRDX1-associated oxidative stress, inflammation, and proliferation in Swiss albino mouse models. Mice were exposed to BaP (50 mg/kg b.wt) orally twice a week for four consecutive weeks and maintained for 16 weeks, respectively. 6-SGL (30 mg/kg b.wt) were orally administered to mouse 1 h before BaP exposure for 16 weeks. After the experiment's termination, 6-SGL (30 mg/kg b.wt) prevented the loss in body weight, increased lung weight, and the total number of tumors in the mice. Moreover, we observed that 6-SGL treatment reverted the activity of BaP-induced lipid peroxidation and antioxidants in mice. Also, 6-SGL impeded the phosphorylation of MAPK family proteins such as Erk1, p38, and Jnk1 in BaP-exposed mice. PRDX1 is an essential antioxidant protein that scavenges toxic radicals and enhances several antioxidant proteins. Overexpression of PRDX1 substantially inhibits MAPKs, proliferation, and inflammation signaling axis. Hence, PRDX1 is thought to be a novel targeting protein for preventing BaP-induced lung cancer. In this study, we have obtained the 6-SGL treatment in a mouse model that reverted BaP-induced depletion of PRDX1 expression. Moreover, pretreatment of 6-SGL (30 mg/kg b.wt) significantly inhibited enhanced proinflammatory cytokines (TNF-α, IL-6, IL-β1, IL-10) and proliferative markers (Cyclin-D1, Cyclin-D2, and PCNA) in BaP-exposed mice. The histopathological studies also confirmed that 6-SGL effectively protected the cells with less damage. Thus, the study demonstrated that 6-SGL could be a potential phytochemical and act as a chemopreventive agent in BaP-induced lung cancer by enhancing PRDX1 expression.

Antioxidant and antimicrobial effects of essential oils from two salvia species with in vitro and in silico analysis targeting 1AJ6 and 1R4U proteins

The Middle Atlas is a Moroccan territory that serves as an abode to plants with incredible biodiversity, of which aromatic and medicinal plants that have been of folkloric use are a significant component. However, their effective utilization in modern medicine requires the characterization of their phytochemicals to facilitate their entry into drug discovery pipelines. Hence, this study aimed to characterize and investigate the antioxidant activity and antimicrobial effects of the essential oils (EOs) of Salvia lavandulifolia subsp. mesatlantica and Salvia officinalis L. by use of in vitro and in silico assays. Ten phytochemicals were identified in the EOs of S. lavandulifolia, while twenty phytochemicals were identified in S. officinalis. Camphor was the most abundant compound in both species, comprising 26.70% and 39.24% of the EOs of S. lavandulifolia and S. officinalis, respectively. The EOs of both plants exhibited significant DPPH free radical scavenging activity, with S. lavandulifolia and S. officinalis showing estimated scavenging rates of 92.97% and 75.20%, respectively. In terms of Ferric Reducing Antioxidant Power (FRAP), S. officinalis demonstrated a higher value (72.08%) compared to that of S. lavandulifolia (64.61%). Evaluation of the antimicrobial effects of the EOs of S. officinalis and S. lavandulifolia against microorganisms revealed bactericidal activities against Proteus mirabilis and Bacillus subtilis at low concentrations. It showed bactericidal activities against Staphylococcus aureus and Candida albicans at a relatively higher concentration. Molecular docking of antioxidant and antimicrobial proteins offers significant insights into ligand–protein interactions, facilitating the development of innovative therapeutics from the current study. Ultimately, this study identified the phytochemical composition of S. lavandulifolia and S. officinalis and highlighted their potential for therapeutic discovery.

Penambahan ekstrak kunyit (Curcuma domestica Val) cair terhadap karakteristik organoleptik dan kandungan antioksidan krispi bayam (crispy spinach)

The human immune system can protect the body from bacteria, viruses, and parasites that cause disease. The immune system becomes more active when enough amounts of macro and micro nutrients are consumed. Antioxidants can also guard against the attack of free radicals and radical chemicals found in the body, limiting the damage caused by the oxidation process. One of them is using turmeric and spinach extract as a supplement. Turmeric extract can be used in the production of crispy spinach cookies. The research was purposed to find out the effect of turmeric extract addition on organoleptic characteristics and antioxidant content crispy spinach. This research used a Experimental research design (Pre-Experimental Design) with three treatments consisting of the addition of 50 ml, 60 ml, and 70 ml of turmeric extract. All three formulations were conducted hedonic tests for know the best formulation, followed by antioxidant content (DPPH method), carbohydrate content (method by different), protein content (Kjedahl), and fat content (Soxhlet). Furthermore, economic analysis and product ranking are carried out. Formulation 3 was chosen because of the highest nutritional content.
 
 Keywords: crispy spinach, immunity, turmeric extract

Prolonged Exposure to High Glucose Induces Premature Senescence Through Oxidative Stress and Autophagy in Retinal Pigment Epithelial Cells.

Chronic hyperglycemia involves persistent high-glucose exposure and correlates with retinal degeneration. It causes various diseases, including diabetic retinopathy (DR), a major cause of adult vision loss. Most in vitro studies have investigated the damaging short-term effects of high glucose exposure on retinal pigment epithelial (RPE) cells. DR is also a severe complication of diabetes. In this study, we established a model with prolonged high-glucose exposure (15 and 75mM exogenous glucose for two months) to mimic RPE tissue pathophysiology in patients with hyperglycemia. Prolonged high-glucose exposure attenuated glucose uptake and clonogenicity in ARPE-19 cells. It also significantly increased reactive oxygen species levels and decreased antioxidant protein (superoxide dismutase 2) levels in RPE cells, possibly causing oxidative stress and DNA damage and impairing proliferation. Western blotting showed that autophagic stress, endoplasmic reticulum stress, and genotoxic stress were induced by prolonged high-glucose exposure in RPE cells. Despite a moderate apoptotic cell population detected using the Annexin V-staining assay, the increases in the senescence-associated proteins p53 and p21 and SA-β-gal-positive cells suggest that prolonged high-glucose exposure dominantly sensitized RPE cells to premature senescence. Comprehensive next-generation sequencing suggested that upregulation of oxidative stress and DNA damage-associated pathways contributed to stress-induced premature senescence of ARPE-19 cells. Our findings elucidate the pathophysiology of hyperglycemia-associated retinal diseases and should benefit the future development of preventive drugs. Prolonged high-glucose exposure downregulates glucose uptake and oxidative stress by increasing reactive oxygen species (ROS) production through regulation of superoxide dismutase 2 (SOD2) expression. Autophagic stress, ER stress, and DNA damage stress (genotoxic stress) are also induced by prolonged high-glucose exposure in RPE cells. Consequently, multiple stresses induce the upregulation of the senescence-associated proteins p53 and p21. Although both apoptosis and premature senescence contribute to high glucose exposure-induced anti-proliferation of RPE cells, the present work shows that premature senescence rather than apoptosis is the dominant cause of RPE degeneration, eventually leading to the pathogenesis of DR.

Coxiella co-opts the Glutathione Peroxidase 4 to protect the host cell from oxidative stress–induced cell death

The causative agent of human Q fever, Coxiella burnetii, is highly adapted to infect alveolar macrophages by inhibiting a range of host responses to infection. Despite the clinical and biological importance of this pathogen, the challenges related to genetic manipulation of both C. burnetii and macrophages have limited our knowledge of the mechanisms by which C. burnetii subverts macrophages functions. Here, we used the related bacterium Legionella pneumophila to perform a comprehensive screen of C. burnetii effectors that interfere with innate immune responses and host death using the greater wax moth Galleria mellonella and mouse bone marrow-derived macrophages. We identified MceF (Mitochondrial Coxiella effector protein F), a C. burnetii effector protein that localizes to mitochondria and contributes to host cell survival. MceF was shown to enhance mitochondrial function, delay membrane damage, and decrease mitochondrial ROS production induced by rotenone. Mechanistically, MceF recruits the host antioxidant protein Glutathione Peroxidase 4 (GPX4) to the mitochondria. The protective functions of MceF were absent in primary macrophages lacking GPX4, while overexpression of MceF in human cells protected against oxidative stress-induced cell death. C. burnetii lacking MceF was replication competent in mammalian cells but induced higher mortality in G. mellonella, indicating that MceF modulates the host response to infection. This study reveals an important C. burnetii strategy to subvert macrophage cell death and host immunity and demonstrates that modulation of the host antioxidant system is a viable strategy to promote the success of intracellular bacteria.

Muscle p62 stimulates the expression of antioxidant proteins alleviating cancer cachexia.

Oxidative stress plays an important role in skeletal muscle atrophy during cancer cachexia, and more glycolytic muscles are preferentially affected. Sequestosome1/SQSTM1 (i.e., p62), particularly when phosphorylated at Ser 349 (Ser 351 in mice), competitively binds to the Kelch-like ECH-associated protein 1 (Keap1) activating Nuclear factor erythroid 2-related factor 2 (Nrf2). Nrf2 then stimulates the transcription of antioxidant/electrophile-responsive elements in target genes. However, a potential role for p62 in the protection of muscle wasting in cachexia remains to be determined. Here, using the well-established cachexia-inducing model of Lewis Lung Carcinoma (LLC) in mice we demonstrate higher expression of antioxidant proteins (i.e., NQO1, HO-1, GSTM1, CuZnSOD, MnSOD, and EcSOD) in the more oxidative and cachexia resistant soleus muscle than in the more glycolytic and cachexia prone extensor digitorum longus muscle. This was accompanied by higher p62 (total and phosphorylated) and nuclear Nrf2 levels in the soleus, which were paralleled by higher expression of proteins known to either phosphorylate or promote p62 phosphorylation (i.e., NBR1, CK1, PKCδ, and TAK1). Muscle-specific p62 gain-of-function (i.e., in p62 mTg mice) activated Nrf2 nuclear translocation and increased the expression of multiple antioxidant proteins (i.e., CuZnSOD, MnSOD, EcSOD, NQO1, and GSTM1) in glycolytic muscles. Interestingly, skeletal muscle Nrf2 haplodeficiency blunted the increases of most of these proteins (i.e., CuZnSOD, EcSOD, and NQO1) suggesting that muscle p62 stimulates antioxidant protein expression also via additional, yet to be determined mechanisms. Of note, p62 gain-of-function mitigated glycolytic muscle wasting in LLC-affected mice. Collectively, our findings identify skeletal muscle p62 as a potential therapeutic target for cancer cachexia.

AR/PCC herb pair inhibits osteoblast pyroptosis to alleviate diabetes-related osteoporosis by activating Nrf2/Keap1 pathway.

Osteoporosis is a prevalent complication of diabetes, characterized by systemic metabolic impairment of bone mass and microarchitecture, particularly in the spine. Anemarrhenae Rhizoma/Phellodendri Chinensis Cortex (AR/PCC) herb pair has been extensively employed in Traditional Chinese Medicine to manage diabetes; however, its potential to ameliorate diabetic osteoporosis (DOP) has remained obscure. Herein, we explored the protective efficacy of AR/PCC herb pair against DOP using a streptozotocin (STZ)-induced rat diabetic model. Our data showed that AR/PCC could effectively reduce the elevated fasting blood glucose and reverse the osteoporotic phenotype of diabetic rats, resulting in significant improvements in vertebral trabecular area percentage, trabecular thickness and trabecular number, while reducing trabecular separation. Specifically, AR/PCC herb pair improved impaired osteogenesis, nerve ingrowth and angiogenesis. More importantly, it could mitigate the aberrant activation of osteoblast pyroptosis in the vertebral bodies of diabetic rats by reducing increased expressions of Nlrp3, Asc, Caspase1, Gsdmd and IL-1β. Mechanistically, AR/PCC activated antioxidant pathway through the upregulation of the antioxidant response protein Nrf2, while concurrently decreasing its negative feedback regulator Keap1. Collectively, our in vivo findings demonstrate that AR/PCC can inhibit osteoblast pyroptosis and alleviate STZ-induced rat DOP, suggesting its potential as a therapeutic agent for mitigating DOP.

Intermediate metabolites and molecular correlates of one‑carbon and nutrient metabolism differ in tissues from Holstein fetuses

Methionine and folate cycles along with transsulfuration comprise the one‑carbon metabolism (OCM) pathway. Amino acids and other nutrients feed into OCM, which is central to cellular function. mRNA abundance, proteins (Western blotting), and metabolites (GC-MC) associated with OCM were used to characterize these mechanisms in fetal tissues. Liver, whole intestine, and semitendinosus muscle were harvested from fetuses in 6 multiparous Holstein cows (37 kg milk/d, 100 d gestation). Data were analyzed using PROC MIXED (SAS 9.4). Protein abundance of BHMT was greatest (P < 0.01) in liver suggesting active remethylation of homocysteine to methionine. This idea was supported by the greater (P < 0.05) mRNA of CBS, BHMT, MTR, SHMT1, and MAT1A (encoding OCM enzymes) in liver. The antioxidant protein GPX3 had greatest (P < 0.05) abundance in liver, whereas the glutathione-transferase GSTM1 was 5-fold greater (P < 0.05) in intestine than liver and muscle. Greatest concentrations of glycine, serine, and taurine along with lower cysteine underscored the relevance of OCM in fetal liver. Phosphoethanolamine concentration was greatest (4-fold, P < 0.05) in intestine and along with the greatest (P < 0.05) mRNA of SLC44A1 (choline transporter), CHKA, and CEPT1 underscored the importance of the CDP-choline pathway. Greatest (P < 0.05) mRNA of PPARA, CPT1A, and HMGCS2 along with lower PCK1 in liver highlighted a potential reliance on fatty acid oxidation. In contrast, greater (P < 0.05) concentration of myo-inositol in muscle and intestine suggested both tissues rely on glucose as main source of energy. Future research should address how environmental inputs such as maternal nutrition alter these pathways in fetal tissues and their phenotypic outcomes.

Determinants of the maximal functional reserve during repeated supramaximal exercise by humans: The roles of Nrf2/Keap1, antioxidant proteins, muscle phenotype and oxygenation

When high-intensity exercise is performed until exhaustion a “functional reserve” (FR) or capacity to produce power at the same level or higher than reached at exhaustion exists at task failure, which could be related to reactive oxygen and nitrogen species (RONS)-sensing and counteracting mechanisms. Nonetheless, the magnitude of this FR remains unknown. Repeated bouts of supramaximal exercise at 120% of VO2max interspaced with 20s recovery periods with full ischaemia were used to determine the maximal FR. Then, we determined which muscle phenotypic features could account for the variability in functional reserve in humans. Exercise performance, cardiorespiratory variables, oxygen deficit, and brain and muscle oxygenation (near-infrared spectroscopy) were measured, and resting muscle biopsies were obtained from 43 young healthy adults (30 males). Males and females had similar aerobic (VO2max per kg of lower extremities lean mass (LLM): 166.7 ± 17.1 and 166.1 ± 15.6 ml kg LLM−1.min−1, P = 0.84) and anaerobic fitness (similar performance in the Wingate test and maximal accumulated oxygen deficit when normalized to LLM). The maximal FR was similar in males and females when normalized to LLM (1.84 ± 0.50 and 2.05 ± 0.59 kJ kg LLM−1, in males and females, respectively, P = 0.218). This FR depends on an obligatory component relying on a reserve in glycolytic capacity and a putative component generated by oxidative phosphorylation. The aerobic component depends on brain oxygenation and phenotypic features of the skeletal muscles implicated in calcium handling (SERCA1 and 2 protein expression), oxygen transport and diffusion (myoglobin) and redox regulation (Keap1). The glycolytic component can be predicted by the protein expression levels of pSer40-Nrf2, the maximal accumulated oxygen deficit and the protein expression levels of SOD1. Thus, an increased capacity to modulate the expression of antioxidant proteins involved in RONS handling and calcium homeostasis may be critical for performance during high-intensity exercise in humans.

Antibiofilm, Antimicrobial, Anti-Quorum Sensing, and Antioxidant Activities of Saudi Sidr Honey: In Vitro and Molecular Docking Studies.

Sidr honey is a valuable source of bioactive compounds with promising biological properties. In the present study, antimicrobial, antioxidant, and anti-quorum sensing properties of Saudi Sidr honey were assessed, along with phytochemical analysis, via gas chromatography-mass spectrometry (GC-MS). In silico study was also carried out to study the drug-likeness properties of the identified compounds and to study their affinity with known target proteins assessed using molecular docking approach. The results showed that Saudi Sidr honey exhibited promising antibacterial activity, with MIC values ranging from 50 to 400 mg/mL and MBC values from 50 to >450 mg/mL. Interestingly, the Saudi Sidr honey was active against Candida auris and Candida neoformans, with an MIC value of about 500 mg/mL. Moreover, the Sidr honey showed important antioxidant activities (ABTS assay: IC50 5.41 ± 0.045 mg/mL; DPPH assay: IC50 7.70 ± 0.065 mg/mL) and β-carotene bleaching test results (IC50 ≥ 20 mg/mL). In addition, the Saudi Sidr honey was able to inhibit biofilm formation on glass slides at 1/2 MIC by 77.11% for Bacillus subtilis, 70.88% for Staphylococcus aureus, 61.79% for Escherichia coli, and 56.64% for Pseudomonas aeruginosa. Similarly, violacein production by Chromobacterium violaceum was reduced by about 56.63%, while the production of pyocyanin by P. aeruginosa was decreased to 46.27% at a low concentration of Saudi Sidr honey. ADMET properties showed that five identified compounds, namely, 1-cyclohexylimidazolidin-2-one, 3-Butyl-3-methylcyclohexanone, 4-butyl-3-methoxy-2-cyclo penten-1-one, 2,2,3,3-Tetramethyl cyclopropane carboxylic acid, and 3,5-dihydroxy-2-(3-methylbut-2-en-1-yl showed promising drug-likeness properties. The compound 3,5-dihydroxy-2-(3-methylbut-2-en-1-yl exhibited the highest binding energy against antimicrobial and antioxidant target proteins (1JIJ, 2VAM, 6B8A, 6F86, 2CDU, and 1OG5). Overall, the obtained results highlighted the promising potential of Saudi Sidr honey as a rich source of bioactive compounds that can be used as food preservatives and antimicrobial, antioxidant, and anti-quorum sensing molecules.

Low-dose Lipopolysaccharide Alleviates Spinal Cord Injury-induced Neuronal Inflammation by Inhibiting microRNA-429-mediated Suppression of PI3K/AKT/Nrf2 Signaling.

This study investigated the impact of low-dose lipopolysaccharide (LPS) on spinal cord injury (SCI) and the potential molecular mechanism. Rats were randomly assigned to four groups: Sham, SCI, SCI + LPS, and SCI + LPS + agomir. Allen's weight-drop method was used to establish an in vivo SCI model. The Basso Bcattie Bresnahan rating scale was employed to monitor locomotor function. An in vitro SCI model was constructed by subjecting PC12 cells to oxygen and glucose deprivation/ reoxygenation (OGD/R). Enzyme-linked immunosorbent assay (ELISA) was applied for the determination interleukin (IL)-1β and IL-6. The dual luciferase reporter assay was used to validate the targeting of microRNA (miR)-429 with PI3K. Immunohistochemical staining was used to assess the expression of PI3K, phosphorylated AKT and Nrf2 proteins. The Nrf2-downstream anti-oxidative stress proteins, OH-1 and NQO1, were detected by western blot assay. MiR-429 expression was detected by fluorescence in situ hybridization and real-time quantitative reverse transcription PCR. In vitro, low-dose LPS decreased miR-429 expression, activated PI3K/AKT/Nrf2, inhibited oxidative stress and inflammation, and attenuated SCI. MiR-429 was found to target and negatively regulate PI3K. Inhibition of miR-429 suppressed low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. In vivo, miR-429 was detectable in neurons. Inhibition of miR-429 blocked low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. Overall, low-dose LPS was found to alleviate SCI-induced neuronal oxidative stress and inflammatory response by down-regulating miR-429 to activate the PI3K/AKT/Nrf2 pathway.

Dragon blood resin ameliorates steroid-induced osteonecrosis of femoral head through osteoclastic pathways.

Objective: Dragon's Blood resin (DBR) is a traditional medicinal substance renowned for its diverse pharmacological effects, which consists of potent anti-inflammatory, antioxidant and angiogenic properties. This study aimed to elucidate its therapeutic mechanism in alleviating steroid-induced osteonecrosis of the femoral head (SIONFH). Methods: Techniques such as SPR and LC-MS were employed to identify and analyze the target proteins of DBR in bone marrow macrophages (BMMs). In vitro, BMMs were treated with RANKL and DBR, and TRAcP staining and actin belt staining were utilized to assess osteoclast activity. The inhibitory effects and underlying mechanisms of DBR on osteoclastogenesis and reactive oxygen species (ROS) generation were determined using real-time PCR, western blotting and immunofluorescence staining. An in vivo SIONFH rat model was set up to assess the curative impacts of DBR using micro-CT scanning and pathological staining. Results: Bioinformatic tools revealed a pivotal role of osteoclast differentiation in SIONFH. Proteomic analysis identified 164 proteins binding in BMMs. In vitro assessments demonstrated that DBR hindered osteoclastogenesis by modulating the expression of specific genes and proteins, along with antioxidant proteins including TRX1 and Glutathione Reductase. Notably, the resin effectively inhibited the expression of crucial proteins, such as the phosphorylation of JNK and the nuclear localization of p65 within the TRAF6/JNK and NFκB signaling pathways. In vivo experiments further confirmed that DBR mitigated the onset of SIONFH in rats by curbing osteoclast and ROS activities. Conclusion: These findings underscore the potential of Dragon's Blood as an effective administration for early-stage SIONFH, shedding light on its therapeutic influence on ROS-mediated osteoclastic signaling pathways.

Differential effects of PGAM5 knockout on high fat high fructose diet and methionine choline-deficient diet induced non-alcoholic steatohepatitis (NASH) in mice

BackgroundPhosphoglycerate mutase 5 (PGAM5), a phosphatase involved in mitochondrial homeostasis, is reported to be closely related to the metabolic stress induced by high-fat diet (HFD) or cold. In this study, we aimed to investigate the effects of PGAM5 on hepatic steatosis, inflammation and fibrosis in nonalcoholic steatohepatitis (NASH).Methods and resultsWe generated PGAM5 global knockout (GKO) mice and their wildtype (WT) littermates using CRISPR/CAS9. The mice were fed with a high fat high fructose (HFHF) diet for 12 weeks or a methionine choline-deficient (MCD) diet (methionine choline supplemented (MCS) as control) for 6 weeks. Hepatic PGAM5 expression was up-regulated in humans with NASH and WT mice fed with HFHF and MCS, and reduced in WT mice fed with MCD diet. In HFHF-fed mice, GKO had reduced body weight, hepatic triglyceride (TG) content and serum transaminase along with decreased hepatic pro-inflammatory and pro-fibrotic responses compared with their WT control. GKO had increased expression of antioxidative gene glutathione peroxidase-6 (GPX6) and activation of mammalian target of rapamycin (mTOR). In mice fed with MCS diet, GKO significantly increased serum TNF-α and IL-6 and decreased hepatic GPX6 mRNA expression. There was no difference in hepatic steatosis, inflammation or fibrosis between GKO and WT mice fed with MCD diet. We investigated the role of PGAM5 deficiency in a variety of cell types. In differentiated THP-1 cells, PGAM5 silencing significantly increased pro-inflammatory cytokine secretion and decreased antioxidative proteins, including nuclear factor erythroid 2- related factors (NRF2), heme oxygenase-1 (HO-1) and GPX6 without affecting mTOR activity. In HepG2 cells with steatosis, PGAM5 knockdown reduced insulin sensitivity, increased mTOR phosphorylation and reduced the expression of NRF2, catalase (CAT), HO-1 and GPX6. Conversely, PGAM5 knockdown reduced TG accumulation, increased insulin sensitivity, and increased antioxidative genes in 3T3-L1 cells, despite the up-regulation in mTOR phosphorylation.ConclusionsPGAM5-KO relieved hepatic steatosis and inflammation in HFHF model, promoted inflammation in MCS-fed mice and had no effects on the MCD-fed model. The distinct effects may be owing to the different effects of PGAM5-KO on anti-oxidative pathways in energy-dependent, possible involves mTOR, and/or cell type-dependent manner. Our findings suggest that PGAM5 can be a potential therapeutic target for NASH.

Vitexin and isovitexin delayed ageing and enhanced stress-resistance through the activation of the SKN-1/Nrf2 signaling pathway

Vitexin and isovitexin, as potential SKN-1/Nrf2 (SKN-1 is a homologous protein of mammalian Nrf2) activators, extended lifespan and promoted healthspan in Caenorhabditis elegans. This study aims to elucidate the role of SKN-1/Nrf2 in vitexin and isovitexin-induced anti-aging and stress-resistance. Vitexin and isovitexin upregulated antioxidant gene and protein expressions, reduced ROS accumulation, and increased SKN-1 accumulation in the nucleus. They prolonged lifespan and clear ROS during stressful conditions in a skn-1-dependent manner. skn-1 was also found to be necessary for these compounds-induced longevity under normal conditions. They were also witnessed to retard cellular senescence and scavenge ROS in senescent cells by directly binding to the pocket of Keap1 to promote the dissociation and activation of Nrf2. This study showed that SKN-1/Nrf2 signaling was vital to delaying ageing and enhancing anti-stress capacity with vitexin and isovitexin. The findings provide new insights into apigenin C-glycosides activating the SKN-1/Nrf2 pathway and demonstrate their potential as candidates for innovative strategies in chemoprophylaxis against ageing and oxidative-related diseases.

Glucocorticoid-Induced Dose-Dependent Reproductive Impairments in Male Albino Rat

The present study is aimed at elucidating the effect of different doses of Dexamethasone (DEX) on testis and epididymis of male Wistar rats. Thirty-six rats were divided into six groups, each containing six rats: one control (CON), and five groups of rats treated with five (30 μg, 40 μg, 50 μg, 60 μg, 70 μg/100 g body weight) doses of dexamethasone intraperitoneally (i.p.) for twenty-one days. The results revealed a dose-dependent decrease in testes and epididymis weight. DEX-treated rats evidenced significant increase in TBARS levels, this being the highest in 70 μg/100 g body weight (bw) which could be a consequence of the highest level of free radical generation. The activity level of antioxidative enzymes Superoxide Dismutase (SOD), Catalase (CAT), Glutathione (GSH), and protein content significantly declined in dexamethasone-treated rats in a dose-dependent manner. Histological observation revealed different degrees of germ cell degeneration/alterations in testis and epididymis The present finding suggests that exposure to dexamethasone (70 μg/100 g bw) can potentially lead to severe impairments in male reproductive tissues (testis and epididymis) structure and function, which may consequently lead to male infertility.

TRIM21-Promoted FSP1 Plasma Membrane Translocation Confers Ferroptosis Resistance in Human Cancers.

Ferroptosis, an iron-dependent form of regulated cell death driven by excessive accumulation of lipid peroxides, has become a promising strategy in cancer treatment. Cancer cells exploit antioxidant proteins, including Ferroptosis Suppressor Protein 1 (FSP1), to prevent ferroptosis. In this study, it is found that the E3 ubiquitin ligase TRIM21 bound to FSP1 and mediated its ubiquitination on K322 and K366 residues via K63 linkage, which is essential for its membrane translocation and ferroptosis suppression ability. It is further verified the protective role of the TRIM21-FSP1 axis in RSL3-induced ferroptosis in cancer cells and a subcutaneous tumor model. Moreover, TRIM21 is highly expressed in multiple gastrointestinal (GI) tumors, and its expression is further stimulated upon ferroptosis induction in cancer cells and the KPC mouse model. In summary, This study identifies TRIM21 as a negative regulator of ferroptosis through K63 ubiquitination of FSP1, which can serve as a therapeutic target to enhance the chemosensitivity of tumors based on ferroptosis induction.

Zexie decoction reduce glucose-dependent lipid accumulation and oxidative stress in Caenorhabditis elegans

BackgroundObesity has become a global public health problem. Zexie decoction (ZXT) is a classic formula from Synopsis of the Golden Chamber. However, the long-term effect of ZXT in lipid accumulation remain unclear. PurposeThis study aims to investigate the effect of ZXT on aging, lipid metabolism and oxidative stress. MethodsDifferent concentration of ZXT was administered to Caenorhabditis elegans (C. elegans) cultured in NGM or the high glucose nematode growth media (GNGM). The lifespan, heat stress resistance, lipid accumulation and related mRNA expression of the worms were examined. Oil Red staining and triglyceride were used to evaluated the lipid accumulation. Nhr-49, fat-5/fat-7, fat-5/fat-6 or skn-1 knockout mutants were used to clarify the effect on lipid metabolism of ZXT. GFP-binding mutants were used to observe the changes in protein expression. ResultsZXT improved the survival rate of C. elegans in lifespan test and heat stress test. ZXT also reduced lipid accumulation in C. elegans and significantly changed the expression of fatty acid synthesis related genes and lipid metabolism related genes. In addition, ZXT-treated C. elegans showed a higher expression of anti-oxidative protein, and reduced the expression of oxidative stress and mitochondrial dysfunction marker. However, when skn-1 was knockdown, ZXT no longer had the effect of maintaining the mitochondria membrane potential and lipid lowering but still effectively decreased the O2·− induced by high glucose. ConclusionsZXT reduced fat accumulation by regulating lipid metabolism via multiple targets and enhanced stress resistance by its antioxidant effect in C. elegans.