Superoxide Dismutase Activity Research Articles

Effects of flavonoid extract from wonderful Kola (Buchholzia Coriacea) seed on antioxidant and liver function biomarkers of fructose fed streptozotocin induced type 2 diabetic albino rats

Wonderful kola is a wonderful herb with excellent healing powers, the seeds is fast gaining reputation because it appears to be a treatment to nearly all ailments. This study evaluates the potential hepatoprotective and antioxidant effects of oral administration of flavonoid-rich seed extract of Buchholzia coriacea seeds in fructose fed Streptozotocin (STZ)-induced diabetic rats. Twenty-five (25) rats were used for this study. They were divided into five (5) groups of five rats each; normal control, negative control (Streptozotocin (STZ)-induced diabetic), positive control (Glibenclamide), 100mg/kg of body weight seed extract and 500mg/kg of body weight seed extract. The flavonoid seed extract of the plant significantly (P<0.05) reduced liver damage caused by STZ, as evidenced by the decreased serum alanine amino transferase and aspartate amino transferase activity, compared to in the group treated with STZ alone. The result of the Superoxide dismutase activity (SOS) shows more antioxidant activity than other antioxidant parameters. However, that of catalase enzyme has the lowest activity as compared to Thiobarbituric acid reactive species (TBARS) and Reduced Glutathione (GSH) activities. All were significantly (P<0.05) improved following the treatments of flavonoid extract. In conclusion, the results show that seeds of Buchholzia coriacea contain a potent antioxidant and hepatoprotective effects, which can give scientific merit to the traditional use of the extract in the management of diabetes and the improvement of its complications.

2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione protects against MPP+-induced neurotoxicity by ameliorating oxidative stress, apoptosis and autophagy in SH-SY5Y cells.

2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) is a cyclohexanedione compound extracted from the roots of Averrhoa carambola L. Several studies have documented its beneficial effects on diabetes, Alzheimer's disease, and cancer. However, its potential neuroprotective effects on Parkinson's disease (PD) have not yet been explored. The present study aimed to investigate the protective effects and underlying mechanisms of DMDD in a cellular model of PD. In this study, SH-SY5Y cells were incubated with or without DMDD following intoxication with the parkinsonian neurotoxin 1-methyl-4-phenylpyridine (MPP+). Cell viability and apoptosis were evaluated using 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and Hoechst 33,342 staining, respectively. The mitochondrial membrane potential (Δψm) was assessed through the JC-10 assay. The activities of superoxide dismutase (SOD) and the levels of reactive oxygen species (ROS) were measured using WST-8 and DCFH-DA assays. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to explore significant biological processes and pathways influenced by DMDD. Molecular docking was employed to predict the domains of potential protein targets interacting with DMDD. Western blotting was subsequently conducted to determine the protein expression levels of TH, Nrf2, Bax, Bcl-2, Caspase-3, Beclin-1, PARP, LC3-II, LC3-I, p-PI3K, PI3K, p-mTOR and mTOR. Our study showed that DMDD treatment significantly increased cell viability and reduced apoptosis in MPP+-treated SH-SY5Y cells. In addition, DMDD treatment reversed the loss of TH expression and Δψm in MPP+-exposed SH-SY5Y cells. Moreover, DMDD treatment reduced MPP+-induced ROS production by promoting SOD activity. Additionally, compared with those in the MPP+ group, the protein expression levels of Beclin-1, Caspase-3, and PARP and the LC3II/I ratio were significantly decreased, whereas the protein expression levels of Nrf2 and the Bcl-2/Bax, p-PI3K/PI3K, and p-mTOR/mTOR ratios were significantly increased in the DMDD-treated group. In conclusion, DMDD protects against MPP+-induced cytotoxicity by mitigating oxidative stress, apoptosis, and autophagy. PI3K/mTOR signaling at least partly mediates the cytoprotective effect of DMDD.

Ameliorative effect of calcium chloride on growth, anatomy and biochemistry of water deficit stressed spinach plant

One of the major problems faced by agricultural sector all over the world is water scarcity. Water deficit stress in plants results in changes in the physiological, morphological, biochemical and molecular traits. Although water deficit stress has been reported to causes change in various parameters of many plants, but neither studied in Indian green leafy vegetable Spinacia oleracea nor have the mitigating effects of calcium chloride treatment been assessed. So, the present project was carried out with the objective to study the effect of water deficit condition on the growth performance of S. oleracea and the effect of calcium chloride treatment on morphology, anatomy, biochemistry and molecular regulation. Seeds of S. oleracea was allowed to germinate and grown for 7 days with normal irrigation and then divided into three groups (Group-1: Plants irrigated with normal conditions, Group-2: Created water deficit stress by low-frequency of water irrigation and Group-3: Water deficit stress induced plants treated with calcium chloride). Some morphological, anatomical and biochemical parameters were affected under the stress condition in S. oleracea and calcium chloride treatment was found to be ineffective. The results revealed that the number of leaves per plant (1.00–2.67), leaf size (1.36 × 0.53 cm), root length (3 cm), chlorophyll content (0.50–0.70 mg/g leaf) and superoxide dismutase activity (51.95 units/100 mg of leaf protein) were significantly decreased in water deficit stress exposed plants when compared to the control. Calcium chloride treatment improved the morphological characters like number of leaves (2.67–4), root length (3.10 cm), soluble sugar content (16.12 mg/g leaf) and anatomical features in S. oleracea plants. These results led to the conclusion that S. oleracea is sensitive to water deficit stress, and that CaCl₂ was moderately effective in mitigating the stress based on majority of the studied parameters. Hence, such treatment strategies could be improved and explored to overcome water deficit stress and also to improve the production of nutritionally important leafy vegetable S. oleracea.

Deciphering the therapeutic effects of Xiyanping injection: insights into pulmonary and gut microbiome modulation, SerpinB2/PAI-2 targeting, and alleviation of influenza a virus-induced lung injury

Infection with Influenza A virus (IAV) induces severe inflammatory responses and lung injury, contributing significantly to mortality and morbidity rates. Alterations in the microbial composition of the lungs and intestinal tract resulting from infection could influence disease progression and treatment outcomes. Xiyanping (XYP) injection has demonstrated efficacy in clinical treatment across various viral infections. However, its specific effects and mechanisms against IAV remain unclear. In this study, we established an IAV infection mice model, and utilized 16 S rRNA sequencing, RNA sequencing, protein chips, and molecular docking, to investigate the mechanisms of XYP injection on altering pulmonary and gut microbiota, and identifying its target sites. We revealed that XYP injection significantly reduced mortality, weight loss, lung viral titers, and lung pathology in IAV-infected mice. XYP injection down-regulated the activity of malondialdehyde, and the levels of interleukin (IL)-1β, IL-5, IL-6, tumor necrosis factor-α, IL-18, IL-15, granulocyte colony-stimulating factor, IL-9, chemokine (C-C motif) ligand-5, and C-X-C motif chemokine ligand 5, while up-regulated the activities of glutathione peroxidase reactive and superoxide dismutase, and the level of interferon-γ. The diversity of the pulmonary and gut microbiota was altered slightly after XYP injection. The linear discriminant analysis of the gut microbes revealed a higher proportion of potentially beneficial bacteria, including Akkermansia, Parabacteroides goldsteinii, Defluviitaleaceae, Oscillospirales, and Eubacterium_coprostanoligenes_group characterized the XYP group. Peritoneal macrophage RNA sequencing highlighted Serpinb2 as the most significantly regulated gene by XYP injection, along with consistent changes in multiple downstream Th2 structure genes. KEGG pathway analysis indicated significant modifications in genes associated with influenza A, mitogen-activated protein kinase signaling, nuclear factor kappa-B signaling, and apoptosis following XYP injection. Finally, human protein chips and molecular docking were carried out to confirm the binding of the main component of XYP injection, andrographolide, with SERPINB2/PAI-2 protein. Overall, our study provides valuable insights into the therapeutic potential of XYP injection in treating influenza, highlighting its multifaceted effects on host microbiota and immune responses, and pinpointing SerpinB2/PAI-2 as the target for XYP injection in exerting anti-inflammatory and antiviral therapeutic mechanisms.

C60 fullerene improves the contractile activity of the injured rat muscle gastrocnemius

The powerful antioxidant properties of C60fullerenes have been widely used in biomedical nanotechnology. Owing to the negative effects of free radicals in oxidative stress processes, antioxidants are required to protect injured muscles. Here, the effect of water-soluble C60fullerenes (daily oral dose 1 mg kg-1) on the process of restoration of contractile activity of skeletal muscle of rats (muscle gastrocnemius) 15 d after the initiation of open trauma of different severity was studied for the first time. The structural organization of C60fullerene nanoparticles in aqueous solution was analyzed by dynamic light scattering and atomic force microscopy techniques. Such biomechanical parameters ofmuscle gastrocnemiuscontraction as integrated muscle power, levels of generation of its maximum and minimum force, and time interval until reaching 50% of the level of force response of the muscle were analyzed. Such biochemical indices as concentrations of c-reactive protein, creatinine, and lactate in the rat blood, as well as indices of pro- and antioxidant balance (activities of superoxide dismutase and catalase, the concentration of reduced glutathione) in the blood and muscle tissue of experimental animals, were investigated. It was found that application of water-soluble C60fullerenes statistically significantly improves biomechanical parameters of contraction of injuredmuscle gastrocnemiusat the level of 30-45 ± 3%, which is confirmed by normalization of biochemical indices in the blood and muscle tissue of rats at the level of 35-50 ± 3% and 20-37 ± 3%, correspondingly, relative to the open injury group. These findings open the possibility of using C60fullerenes as potential therapeutic nanoagents capable of correcting pathological states of the muscular system during the physiological repair of open injuries.

Proteomic Analysis Is Needed to Understand the Vulnerability of Bunodosoma cavernatum Sea Anemones to Climate Change.

Sea anemones play a crucial role in marine ecosystems. Recent studies have highlighted their physiological and ecological responses to thermal stress. Therefore, our objective was to perform a proteomic analysis of Bunodosoma cavernatum sea anemones in the Gulf of Mexico, subjected to thermal stress, to understand whether these organisms activate specific processes to resist increased temperature. We submitted one group of sea anemones to variable temperatures (26 to 32 °C) and another group to a constant temperature (28 °C) for 1.5 months. Then we subjected them to thermal stress (32 °C) for 2 weeks. We evaluated the enzymatic activity and proteome in the columns and tentacles. The main effect of the temperature regime change is a reduction in mass. Also, sea anemones synthesized proteins related to the activation of the immune system and protection against temperature. We observed decreased peroxidase activity, while superoxide dismutase activity was higher only in the constant temperature group. On the basis of these data, we deduce that B. cavernatum sea anemones are vulnerable to climate change because they stop producing toxins in their tentacles when faced with thermal stress and activate cellular responses that make them susceptible to pathogens. These responses are not sufficient to guarantee an optimal health state.

Effects of copper/graphene oxide core-shell nanoparticles on Rhipicephalus ticks and their detoxification enzymes

Nanopesticides have been recently introduced as novel pesticides to overcome the drawbacks of using traditional synthetic pesticides. The present study evaluated the acaricidal activity of Copper/Graphene oxide core-shell nanoparticles against two tick species, Rhipicephalus rutilus and Rhipicephalus turanicus. The Copper/Graphene oxide core-shell nanoparticles were synthetized through the solution plasma (SP) method under different conditions. The nanoparticles synthesized at 180 W and 45 min were highly toxic to Rh. rutilus and Rh. turanicus, with 50% lethal concentration (LC50) values of 248.1 and 195.7 mg ml−1, respectively, followed by those which were synthesized at 120 W/30 mins (LC50 = 581.5 and 526.5 mg ml−1), 120 W/15 mins (LC50 = 606.9 and 686.7 mg ml−1), and 100/45 mins (LC50 = 792.9 and 710.7 mg ml−1), after 24 h of application. The enzyme assays revealed that 180 W/45 min treatment significantly inhibited the activity of acetylcholinesterase (115 ± 0.81 and 123 ± 0.33 U/ mg protein/min) and superoxide dismutase (290 ± 0.18 and 310 ± 0.92 U/ mg protein/min) in Rh. rutilus and Rh. turanicus, respectively, as compared with the negative control. The results also revealed a significantly increased catalase activity (895 ± 0.37 and 870 ± 0.31 U/ mg protein/min) in Rh. rutilus and Rh. turanicus, respectively. The above results indicated that Copper/Graphene oxide core-shell nanoparticles could be a promising alternatives for the management of ticks.

Investigating the effect of marigold flower extract on bioactive compounds, antioxidants and postharvest quality and shelf life of peach during storage

Abstract Peaches are a climacteric and highly perishable fruit with a short shelf life under ambient storage conditions which hampers further marketing and leads often to economic losses. Consequently, searching for pre-storage treatments to maintain shelf life and preserve the quality of peach fruit is an important task. Therefore, the present study was designed to investigate the efficiency of marigold flower extract (MFE) application on the shelf life and some quality attributes of peach fruit under ambient storage conditions. Peach fruit of cv. Early grand was treated with different concentrations of MFE (control, 10%, 20%, and 30%) and stored at 25 ± 2 °C for 8 days. The results revealed that at the end of the storage period, MFE (30%) suppressed the decrease of weight loss (21.51%), decay incidence (10.74%), solid-acid ratio (20.91%), pH (4.57), MDA (2.23 nmol/g FW) and H2O2 (3.01 nmol/g FW) compared to the control. Moreover, MFE (30%) increased the fruit firmness (93.93 N), soluble solid content (11.97 Brix), titratable acidity (0.57%), total sugar (10.01%), reducing sugars (5.15%), ascorbic acid (29.53 mg/100 mL), the total phenolic content (130.26 mg GAE/100 g FW) as well as the superoxide dismutase (309.79 U/mg), peroxidase (42.65 U/mg) and catalase (56.30 U/mg) enzyme activity compared to the untreated peach fruits. The results of our study suggested that the application of MFE particularly at 30% might be employed commercially as an environment-friendly approach to delay postharvest senescence and maintain quality attributes of peach fruit under ambient conditions.

HYQTD Drug-containing Serum Alleviates H2O2-induced Endothelial Oxidative Damage by Increasing Mitochondrial ATP Synthesis and Inhibiting ROS.

Atherosclerosis (AS) is caused by the endothelium injury associated with oxidative stress. Previous studies have shown that the Phlegm-Eliminating and Stasis- Transforming Decoction (Huayu Qutan Decoction, HYQTD) has mitochondrial protective function. The objective of this research was to explore how HYQTD drug-containing serum (HYQTD-DS) could potentially protect mitochondrial energy production in endothelial cells (ECs) from injury caused by hydrogen peroxide (H2O2)-induced oxidative damage in AS through SIRT1/PGC-1α/ Nrf2 pathway. After preparation of containing serum, the cells were divided into various categories, such as control group, H2O2 group (an oxidative damage model), HYQTD group, Selisistat (EX527, a SIRT1 inhibitor) combined with H2O2 group, and EX527 combined with HYQTD group. The evaluation of oxidative stress involved measuring reactive oxygen species (ROS) and malondialdehyde (MDA) generation, as well as Superoxide Dismutase (SOD) activity. Mitochondrial function and ultrastructure were measured by Transmission electron microscopy (TEM), mitochondrial membrane potential (MMP), rate of oxygen consumption (OCR), respiratory chain complex activities, and ATP production. The key proteins and gene levels in the SIRT1/PGC-1α/Nrf2 pathway was quantified by quantitative real-time PCR (RT-PCR) and Western blotting analysis. We found oxidative stress, mitochondrial damage, and mitochondrial energy disorder in H2O2-induced ECs. However it indicated a marked reversal after pretreated with HYQTD-DS. Mechanistically, EX527 induced increased oxidative stress, worse mitochondrial dysfunction, and less ATP synthesis. We demonstrated that HYQTD-DS attenuated oxidative stress, improved mitochondrial function, and up-regulated mitochondrial ATP synthesis by activating SIRT1/PGC- 1α/Nrf2 pathway-induced mitochondrial biogenesis and its downstream NADH dehydrogenase (ubiquinone) flavoprotein 2 (NDV2).

Effects of Silibinin on Delaying Aging in Drosophila melanogaster

Aging is an inevitable physiological process, but delaying aging has always been an enduring human pursuit. Silibinin (SIL), derived from the seeds of the milk thistle plant, exhibits a broad spectrum of pharmacological properties, including anti-tumor effects, liver protection, inhibition of apoptosis, and alleviation of inflammation. However, whether it has anti-aging effects remains unclear. The SIL dietary supplement to Drosophila melanogaster prolonged lifespan, improved climbing ability, ameliorated age-associated intestinal barrier disruption, enhanced the resistance to oxidative stress, and increased the enzyme activities of superoxide dismutase (SOD) and catalase (CAT). Furthermore, RNA-seq results showed that SIL addition significantly upregulated 74 genes and downregulated 50 genes compared with the control. KEGG (Kyoto Encyclopedia of genes and genomes) analysis demonstrated that these differentially expressed genes were primarily involved in the Toll signaling pathway and endoplasmic reticulum proteins processing, six among which, including IM2, IM3, Drsl3, CG7556, GCS1, and TRAM, were particularly involved in the regulation by SIL supplementation. The results indicate that SIL exhibits anti-aging effects by enhancing antioxidant capacity and regulating aging-related signaling pathways. Therefore, SIL shows a potential application in anti-aging dietary regimens.

HC-A solution limb perfusion alleviates liver damage induced by limb ischemia-reperfusion injury in pigs.

The aim of the study was to explore a feasible method for alleviating limb ischemia-reperfusion injury (LI/RI) through the use of a high-concentration citrate solution (HC-A solution) for limb perfusion (LP). Eighteen pigs were divided into three groups: the Sham group, LI/RI group, and HCA group. The Sham group underwent exposure of the iliac artery and vein. The LI/RI group underwent tourniquet placement and clamping of the iliac artery and vein to simulate LI/RI. The HCA group received HC-A solution LP for 30 min through the left iliac artery below the level of blood flow occlusion based on the LI/RI group. Oxidative stress markers and inflammatory response markers were compared among the three groups. Compared to the LI/RI group, the HCA group showed significantly lower levels of serum creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), and significantly greater activities of serum superoxide dismutase (SOD) (p < 0.05). There were no significant differences in serum interleukin-6 (IL-6) or in muscle MDA, SOD, TNF-α, and IL-6 between the HCA group and the LI/RI group (p > 0.05). Compared to the LI/RI group, MDA, TNF-α, and IL-6 levels in the liver were significantly lower in the HCA group (p < 0.05), while SOD activities were not significantly different (p > 0.05). Histopathological examination revealed reduced skeletal muscle and liver damage in the HCA group compared to the LI/RI group. HC-A solution LP can alleviate liver damage caused by LI/RI in pigs.

Dihydromyricetin Protects Against Hypoxia/Reoxygenation Injury in Cardiomyocytes by Activating miR-34a-Mediated Notch1 Pathway.

Dihydromyricetin (Dih), a naturally occurring flavonoid, has been identified to exert a protective effect against ischemia/reperfusion injury. However, the detailed mechanisms remain unclear. Here we investigated the biological role of Dih in preventing hypoxia/reoxygenation (H/R) injury in cardiomyocytes. The results showed that Dih protected cardiomyocytes against H/R-induced apoptosis, as proved by improved cell viability and decreased lactate dehydrogenase (LDH) release, cell apoptosis percentage, and caspase-3/7 activity. H/R-induced oxidative stress in cardiomyocytes was also prevented by Dih with increased activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT), and decreased levels of malondialdehyde (MDA) and reactive oxygen species (ROS). Treatment with Dih prevented H/R-induced increase in the activities of myocardial enzymes aspartate aminotransferase (AST), creatine kinase-MB (CK-MB), and creatine kinase (CK). miR-34a expression was upregulated after H/R stimulation, which could be attenuated by Dih pretreatment. Besides, miR-34a overexpression attenuated the protective effects of Dih against H/R-caused increase in apoptosis, oxidative stress, and myocardial enzyme activities. Next, we demonstrated that Notch1 was a target molecule of miR-34a. Notch1 overexpression reversed the role of miR-34a in regulating the cardioprotective effect of Dih on H/R injury. These observations indicated that the cardioprotective effect of Dih against H/R injury was mediated by the miR-34a/Notch1 signaling. Dih may be a candidate agent for improving the clinical efficacy of cardiac ischemia/reperfusion injury treatment.

Enhancing Probiotic, Sensory, and Antioxidant Properties of Polygonatum sibiricum Red. and Crataegus pinnatifida Bunge through Lactic Acid Fermentation for Functional Food Development

Abstract This study explores the use of Polygonatum sibiricum Red. and Crataegus pinnatifida Bunge as edible-medicine homology ingredients through lactic acid fermentation to enhance probiotic content, sensory quality, and antioxidant properties. Using a mixed culture of Lactobacillus plantarum AS1.2437 and Lactobacillus casei CICC21019, the fermentation significantly increased lactic acid bacteria counts, improved taste, and boosted antioxidant activities, particularly in reducing Fe3+ and scavenging DPPH and ABTS free radicals (P &amp;lt; 0.05). During fermentation, changes in reducing sugars, total acids, superoxide dismutase (SOD) activity, and amino acid compositions were monitored. Results showed significant increases in total phenols, flavonoids, SOD activity, and polysaccharide content (P &amp;lt; 0.05). Essential amino acids increased by 4.86%, and therapeutic amino acids rose from 50.88% to 53.59%. The Boltzmann model effectively captured these changes, achieving a high coefficient of determination (R2 &amp;gt; 0.99), indicating strong predictive accuracy. Key organic acids like lactic, acetic, and tartaric acids contributed to enhanced antioxidant activity. This research provides a comprehensive understanding of the physicochemical, medicinal, and antioxidant properties of the fermented Polygonatum sibiricum Red. and C. pinnatifida Bunge (PsR.-CpB.) enzyme, establishing a theoretical foundation for developing functional foods aligned with the edible-medicine homology concept.

Pharmacological modulation of Sigma-1 receptor ameliorates pathological neuroinflammation in rats with diabetic neuropathic pain via the AKT/GSK-3β/NF-κB pathway.

Diabetic neuropathic pain (DNP) is a common complication of diabetes mellitus (DM) and is characterized by spontaneous pain and neuroinflammation. The Sigma-1 receptor (Sig-1R) has been proposed as a target for analgesic development. It is an important receptor with anti-inflammatory properties and has been found to regulate DNP. However, it is not known whether Sig-1R can ameliorate pathological neuroinflammation in DNP. The present study used a rat model of DNP and a highly selective agonist of Sig-1R to assess the effects of the protein on neuropathic pain in rats with type 2 diabetes mellitus. The rats were divided into Control, Model, Sig-1R agonist PRE-084 (0.3, 0.6, 1mg/kg), and metformin (Met, 20mg/kg) groups, with seven rats per group, and their body weight, fasting blood glucose, mechanical withdrawal threshold and thermal withdrawal latency were tested weekly for two weeks. After treatment with PRE-084, the pain thresholds in the DNP rats were significantly improved, together with pathological changes in the dorsal root ganglion, reductions in the serum levels of TNF-α, IL-1β, IL-6, MOD, and prostaglandin E2 (PGE2), and the activity of superoxide dismutase was increased. The mRNA levels of TNF-α, IL-1β, and cyclooxygenase 2 (COX-2) were reduced. Pharmacological inhibition of Sig-1R with BD1047 (10μM) abolished Sig-1R-mediated activation of lipopolysaccharide-treated BV-2 microglial cells. It was also found that PRE-084 increased phosphorylation of serine/threonine protein kinase B (AKT) and glycogen synthase kinase 3β (GSK-3β) at Ser9, inhibiting nuclear factor kappa B (NF-κB)-mediated neuroinflammation in the dorsal root ganglion, thus reducing DNP. The findings suggest that the effect of Sig-1R agonist PRE-084 on DNP may reduce the level of inflammation through the up-regulation of AKT/GSK-3β and down-regulation of the NF-κB signaling, thereby contributing to the treatment of the disease.

Relaxin‐2 Alleviated Ferroptosis and Inflammation in TNF‐α‐Treated Chondrocytes via Regulating the Nrf2/NF‐κB Signaling Pathway

ABSTRACTExcessive inflammatory responses and ferroptosis‐associated dysfunction of chondrocytes are significant pathological features of osteoarthritis (OA). Relaxin‐2, a well‐known hormone involved in metabolic regulation across various tissues and cells, has not been extensively studied in the context of chondrocyte dysfunction and OA. T/C‐28a2 cells were stimulated with tumor necrosis factor α (TNF‐α) (10 ng/mL) with or without recombinant human relaxin‐2 (rh relaxin‐2) (25, 50 nM) for 48 h. Various techniques, including 2′,7′‐dichlorodihydrofluorescein diacetate (DCFH‐DA) staining, real‐time PCR, western blot analysis, enzyme‐linked immunosorbent assay (ELISA), and luciferase activity assays, were employed. We discovered that exposure to TNF‐α decreased the expression of Relaxin‐2 at both the mRNA and protein levels. Further investigation revealed that treatment with rh relaxin‐2 mitigated lipid peroxidation by reducing levels of reactive oxygen species (ROS), malondialdehyde (MDA), and 4‐hydroxy‐2‐nonenal (4‐HNE), while increasing superoxide dismutase (SOD) activity and glutathione (GSH) content. Notably, rh relaxin‐2 restored iron metabolism balance disrupted by TNF‐α in human T/C‐28a2 chondrocytes by decreasing Fe2+ levels, downregulating transferrin receptor 1 (TFR1) gene expression, and upregulating ferritin gene expression. Additionally, rh relaxin‐2 alleviated ferroptosis induced by TNF‐α in these cells by increasing glutathione peroxidase 4 (GPX4) expression, decreasing acyl‐CoA synthetase long‐chain family member 4 (ACSL4) expression, and reducing lactate dehydrogenase (LDH) release. Rh relaxin‐2 also inhibited the TNF‐α‐induced expression of pro‐inflammatory cytokines, interleukin‐1β (IL‐1β) and interleukin‐6 (IL‐6). Interestingly, rh relaxin‐2 attenuated TNF‐α‐induced chondrocyte degeneration by suppressing matrix metalloproteinase‐13 (MMP‐13) expression and preventing degradation of collagen type II alpha1 chain (Col2α). Mechanistically, these effects of rh relaxin‐2 were found to be mediated through the nuclear factor erythroid 2‐related factor 2 (Nrf2)/NF‐κB signaling pathway. Our findings suggest that rh relaxin‐2 could be a potential therapeutic strategy for OA.

Interactive Effects of Dichlorvos, Dimethoate, And Cypermethrin On Growth and Oxidative Homeostasis in Maize (Zea Mays)

This study investigates the effects of dichlorvos, dimethoate, and cypermethrin, individually and in combinations, on maize (Zea mays) growth and physiology. Maize seeds were grown in pesticide-treated soil, and growth parameters, oxidative stress markers, and enzymatic activities were evaluated. Growth was significantly inhibited by pesticide exposure, with Group H (triple pesticide combination) showing the most severe reduction in plant height (48% lower) and stem girth (40% lower) compared to the control (p &lt; 0.05). Biochemical assays revealed significant declines in catalase (CAT) and superoxide dismutase (SOD) activities across all tissues, indicating compromised oxidative stress defenses. For instance, CAT activity in roots decreased by 65% in Group H compared to the control (p &lt; 0.05). Concurrently, malondialdehyde (MDA) concentrations, a marker of lipid peroxidation, increased significantly, with Group H showing a 75% rise in leaves relative to the control (p &lt; 0.05). Relative water content (RWC) also decreased substantially, with Group H recording the lowest hydration levels (37% reduction, p &lt; 0.05). These findings suggest synergistic or cumulative toxicity from combined pesticide exposure, with the most pronounced effects observed in triple pesticide treatments. Statistical analyses (ANOVA, Tukey's test) confirmed significant differences across groups (p &lt; 0.05), reinforcing the reliability of the results. This study highlights the adverse impacts of pesticide combinations on maize growth and physiological stability, emphasizing the need for sustainable pest management strategies, such as integrated pest management (IPM), to mitigate the ecological and agricultural risks of pesticide use.

Synergistic Role of Streptomyces Composite Inoculants in Mitigating Wheat Drought Stress Under Field Conditions

Wheat (Triticum aestivum L.) is a globally important staple crop; however, its growth and yield are severely limited by drought stress. This study evaluated the effects of a combined microbial inoculant, Streptomyces pactum Act12 and Streptomyces rochei D74, on wheat photosynthesis, physiological traits, and yield under drought conditions. Key physiological and yield parameters were measured during the jointing, heading, and grain-filling stages. Drought stress significantly reduced chlorophyll content, maximum photochemical efficiency of photosystem II (PSII) (Fv/Fm), and antioxidant enzyme activities, while increasing malondialdehyde (MDA) levels, leading to a notable yield decline. In contrast, inoculation with Streptomyces strains alleviated these adverse effects, with the combined inoculant (Act12+D74) group demonstrating the most significant improvement. Chlorophyll content increased by up to 32.60%, Fv/Fm improved by 43.07%, and antioxidant enzyme activities were enhanced, with superoxide dismutase (SOD) activity increasing by 19.32% and peroxidase (POD) activity by 75.44%. Meanwhile, MDA levels were reduced by 61.61%. The proline content in the combined inoculant group increased by 90.44% at the jointing stage and the soluble protein content increased by 60.17% at the heading stage. Furthermore, it improved the yield by 26.19% by increasing both effective spikes and grains per spike. For the first time, this study revealed the synergistic effects of Act12 and D74 in enhancing photosynthesis, strengthening antioxidant defenses, and optimizing osmotic regulation under drought conditions. These findings provide a theoretical basis for developing environmentally friendly drought management strategies and highlight the potential applications of this inoculant in sustainable agriculture.

Soybean Flour Fortified with Gryllus assimilis Powder to Increase Iron Bioavailability Improves Gut Health and Oxidative Balance In Vivo

Background: Insects like Gryllus assimilis have an excellent nutritional profile, including iron. However, the bioavailability of this iron and its effects on intestinal health and oxidative balance remain unclear. To enhance acceptance, insects can be used in powder form and combined with common flours. Objective: This study evaluates the effects of Gryllus assimilis powder, alone or with soy flour, on iron bioavailability, intestinal health, and oxidative balance in rodents. Methods: Using the hemoglobin depletion/repletion method, 32 male Wistar rats were divided into four groups: A (standard diet + ferrous sulfate), B (diet + Gryllus assimilis + soy flour), C (diet + Gryllus assimilis), and D (diet + soy flour). Hemoglobin levels, regeneration efficiency, biological value, serum markers, intestinal health, and oxidative balance were assessed. Results: Food intake, weight gain, and bioavailability measures showed no differences. However, the Gryllus + soy group showed higher weekly and final hemoglobin levels than Gryllus alone. This combination also improved acetic acid levels, fecal moisture, and oxidative balance, increasing superoxide dismutase activity while reducing peroxidation products compared to Gryllus alone. Conclusion: These findings highlight the potential benefits of combining Gryllus assimilis with soy flour for iron bioavailability and overall health.

Sly-miR398 Participates in Heat Stress Tolerance in Tomato by Modulating ROS Accumulation and HSP Response

Heat stress is one of the most important environmental problems in agriculture, which severely restricts the growth and yield of plants. In plants, microRNA398 (miR398) negatively regulates the activity of superoxide dismutase (SOD) by modulating the expression of its coding genes (CSDs) post-transcriptionally, thereby regulating reactive oxygen species (ROS) homeostasis and stress resistance. In this study, the role of miR398 in heat stress tolerance in tomatoes was investigated. Under heat stress, the expression of miR398 was upregulated in tomatoes, while the expression of its target genes (CSD1 and CSD2) and SOD activity was downregulated. Furthermore, by comparing the heat stress response in wild type (WT) and a transgenic line overexpressing MIR398 (miR398-OE), the results showed that overexpression of miR398 promoted tomato growth and the expression of genes encoding heat shock factor (HSF, transcription factor) and heat shock protein (HSP) under heat stress. Meanwhile, downregulated activity of antioxidant enzymes, including SOD, catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX), and enhanced ROS accumulation was observed in miR398-OE compared with that in WT under heat stress. Further study using dimethylthiourea (DMTU, a ROS scavenger) indicated that the enhanced plant growth and expression of HSFs/HSPs was based on the promoted accumulation of ROS in miR398-OE. Overall, the results of this study revealed that the upregulated expression of miR398 in response to heat stress would modulate the antioxidant system and enhance ROS accumulation, thereby enhancing the expression of HSFs and HSPs and heat stress tolerance in tomatoes.

Uncovering the molecular mechanisms of Acer fabri in adjusting to low-temperature stress through integrated physiological and transcriptomic analysis

Acer fabri is an excellent color-leaf tree species, with high ornamental value. Low temperatures are known to limit the growth and geographical distribution of A. fabri. The molecular mechanism of A. fabri in response to low-temperature stress was rarely reported. To understand the molecular mechanism of A. fabri in response to low-temperature stress, relevant physiological changes were identified and the transcriptome sequencing was conducted under different stress durations. The results showed that the proline, the soluble sugar (SS) and the soluble protein (SP) content increased in A. fabri leaves under low-temperature stress, while the peroxidase (POD) and activating superoxide dismutase (SOD) activity increased first and then decreased. It was also found by the OPLS-DA analysis that SOD is the most important physiological indicator of A. fabri in response to low-temperature stress. By transcriptome sequencing, a total of 56,732 genes were identified, including 832 transcription factors (TFs). Differentially expressed genes (DEGs) were significantly enriched in metabolic pathways, phytohormone signaling, and plant mitogen-activated protein kinase (MAPK) signaling pathways. Moreover, the analysis of gene co-expression networks, specifically weighted gene co-expression network analysis (WGCNA), indicates that Af0048792 and Af0026061 could be significant in the response to stress from low temperatures. Furthermore, it was observed that NAC (Af0033429) and MIKC (Af0004917) might have interactions with Af0048792, and MIKC (Af0004917) may additionally interact with Af0026061. These findings could enhance our understanding of the molecular mechanisms of A. fabri in response to low-temperature stress.