Reactive Oxygen Species Scavenging Enzymes Research Articles

Exogenous expression of PGC-1α during in vitro maturation impairs the developmental competence of porcine oocytes

Objectives of the current study were to examine the effects of exogenous expression of PGC-1α, which is a transcription factor responsive for controlling mitochondrial DNA (mtDNA) replication, mitochondria quantity control, mitochondrial biogenesis, and reactive oxygen species (ROS) maintenance, in porcine oocytes during in-vitro maturation (IVM) on the developmental competence, as well as mitochondrial quantity and function. Exogenous over-expression of PGC-1α by injection of the mRNA construct into oocytes 20 h after the start of IVM culture significantly increased the copy number of mtDNA in the oocytes, but reduced the incidences of oocytes matured to the metaphase-II stage after the IVM culture for totally 44 h and completely suppressed the early development in vitro to the blastocyst stage following parthenogenetic activation. The exogenous expression of PGC-1α also significantly induced spindle defects and chromosome misalignments. Furthermore, markedly higher ROS levels were observed in the PGC-1α-overexpressed mature oocytes, whereas mRNA level of SOD1, encoded for a ROS scavenging enzyme, was decreased. These results conclude that forced expression of PGC-1α successfully increase mtDNA copy number but led to increased ROS production, evidently by downregulation of SOD1 gene expression, inducement of spindle aberration/chromosomal misalignment, and consequently reduction in the meiotic and developmental competences of porcine oocytes.

Hierarchical multi-task deep learning-assisted construction of human gut microbiota reactive oxygen species-scavenging enzymes database.

In the process of oxygen reduction, reactive oxygen species (ROS) are generated as intermediates, including superoxide anion (O2-), hydrogen peroxide (H2O2), and hydroxyl radicals (OH-). ROS can be destructive, and an imbalance between oxidants and antioxidants in the body can lead to pathological inflammation. Inappropriate ROS production can cause oxidative damage, disrupting the balance in the body and potentially leading to DNA damage in intestinal epithelial cells and beneficial bacteria. Microorganisms have evolved various enzymes to mitigate the harmful effects of ROS. Accurately predicting the types of ROS-scavenging enzymes (ROSes) is crucial for understanding the oxidative stress mechanisms and formulating strategies to combat diseases related to the "gut-organ axis." Currently, there are no available ROSes databases (DBs). In this study, we propose a systematic workflow comprising three modules and employ a hierarchical multi-task deep learning approach to collect, expand, and explore ROSes-related entries. Based on this, we have developed the human gut microbiota ROSes DB (http://39.101.72.186/), which includes 7,689 entries. This DB provides user-friendly browsing and search features to support various applications. With the assistance of ROSes DB, various communication-based microbial interactions can be explored, further enabling the construction and analysis of the evolutionary and complex networks of ROSes DB in human gut microbiota species.IMPORTANCEReactive oxygen species (ROS) is generated during the process of oxygen reduction, including superoxide anion, hydrogen peroxide, and hydroxyl radicals. ROS can potentially cause damage to cells and DNA, leading to pathological inflammation within the body. Microorganisms have evolved various enzymes to mitigate the harmful effects of ROS, thereby maintaining a balance of microorganisms within the host. The study highlights the current absence of a ROSes DB, emphasizing the crucial importance of accurately predicting the types of ROSes for understanding oxidative stress mechanisms and developing strategies for diseases related to the "gut-organ axis." This research proposes a systematic workflow and employs a multi-task deep learning approach to establish the human gut microbiota ROSes DB. This DB comprises 7,689 entries and serves as a valuable tool for researchers to delve into the role of ROSes in the human gut microbiota.

Selenium Nanoparticles as Neuroprotective Agents: Insights into Molecular Mechanisms for Parkinson's Disease Treatment.

Oxidative stress and the accumulation of misfolded proteins in the brain are the main causes of Parkinson's disease (PD). Several nanoparticles have been used as therapeutics for PD. Despite their therapeutic potential, these nanoparticles induce multiple stresses upon entry. Selenium (Se), an essential nutrient in the human body, helps in DNA formation, stress control, and cell protection from damage and infections. It can also regulate thyroid hormone metabolism, reduce brain damage, boost immunity, and promote reproductive health. Selenium nanoparticles (Se-NPs), a bioactive substance, have been employed as treatments in several disciplines, particularly as antioxidants. Se-NP, whether functionalized or not, can protect mitochondria by enhancing levels of reactive oxygen species (ROS) scavenging enzymes in the brain. They can also promote dopamine synthesis. By inhibiting the aggregation of tau, α-synuclein, and/or Aβ, they can reduce the cellular toxicities. The ability of the blood-brain barrier to absorb Se-NPs which maintain a healthy microenvironment is essential for brain homeostasis. This review focuses on stress-induced neurodegeneration and its critical control using Se-NP. Due to its ability to inhibit cellular stress and the pathophysiologies of PD, Se-NP is a promising neuroprotector with its anti-inflammatory, non-toxic, and antimicrobial properties.

Regulation of plum fruit cracking process during cold storage period by treatment of preharvest chitosan

Cracking fruit is a serious quality change phenomenon in the storage and transportation of postharvest plum fruit. To reduce the cracking rate of plum fruit during cold storage, the 'Guofeng No.7' plum was sprayed with chitosan with 1.5% concentration 30d from preharvest. The postharvest plum fruit was stored at (0±0.5) °C and (85-90) % relative humidity. The results showed that chitosan treatment could reduce the reactive oxygen species (ROS) level of plum fruit, effectively delaying the fruit cracking. Compared with the control, chitosan treated fruits inhibited the increase of free radical content such as O2-· and H2O2 and delayed the increase of membrane permeability and the accumulation of MDA (from 71.9 mmol/g to 53.5 mmol/g). It also regulates lipoxygenase (LOX), peroxidase (POD), superoxide dismutase (SOD), catalase (CAT) (enzymatic activity from 34.15 g/min to 42.21 g/min) and ascorbate peroxidase (APX) activity and gene expression during fruit cracking to maintain membrane integrity and intracellular compartmentalization, thereby increasing the crack tolerance of plum fruit during cold storage. These results indicate that preharvest chitosan treatment can inhibit the metabolism of active oxygen species, increase the activity of ROS scavenging enzymes and gene expression, and delay the postharvest cracking of plum fruit.

Alleviating chromium-induced oxidative stress in Vigna radiata through exogenous trehalose application: insights into growth, photosynthetic efficiency, mineral nutrient uptake, and reactive oxygen species scavenging enzyme activity enhancement

Trehalose serves as a crucial osmolyte and plays a significant role in stress tolerance. The influence of exogenously added trehalose (1 and 5 mM) in alleviating the chromium (Cr; 0.5 mM) stress-induced decline in growth, photosynthesis, mineral uptake, antioxidant system and nitrate reductase activity in Vigna radiata was studied. Chromium (Cr) significantly declined shoot height (39.33%), shoot fresh weight (35.54%), shoot dry weight (36.79%), total chlorophylls (50.70%), carotenoids (29.96%), photosynthesis (33.97%), net intercellular CO2 (26.86%), transpiration rate (36.77%), the content of N (35.04%), P (35.77%), K (31.33%), S (23.91%), Mg (32.74%), and Ca (29.67%). However, the application of trehalose considerably alleviated the decline. Application of trehalose at both concentrations significantly reduced hydrogen peroxide accumulation, lipid peroxidation and electrolyte leakage, which were increased due to Cr stress. Application of trehalose significantly mitigated the Cr-induced oxidative damage by up-regulating the activity of reactive oxygen species (ROS) scavenging enzymes, including superoxide dismutase (182.03%), catalase (125.40%), ascorbate peroxidase (72.86%), and glutathione reductase (68.39%). Besides this, applied trehalose proved effective in enhancing ascorbate (24.29%) and reducing glutathione content (34.40%). In addition, also alleviated the decline in ascorbate by Cr stress to significant levels. The activity of nitrate reductase enhanced significantly (28.52%) due to trehalose activity and declined due to Cr stress (34.15%). Exogenous application of trehalose significantly improved the content of osmolytes, including proline, glycine betaine, sugars and total phenols under normal and Cr stress conditions. Furthermore, Trehalose significantly increased the content of key mineral elements and alleviated the decline induced by Cr to considerable levels.

Phytic acid-mediated enhancement of Meyerozyma caribbica biocontrol of Aspergillus carbonarius infection in grape berries through regulation of ROS metabolism

Sour rot, an infection caused by Aspergillus carbonarius is one of the detrimental diseases that damage grape vines after harvest. To investigate the biocontrol of this mycotoxigenic fungus, the current study explored the efficiency of Meyerozyma caribbica pretreated with phytic acid (PA) in reducing sour rot in table grapes through the regulation of reactive oxygen species (ROS) production. The content of hydrogen peroxide (H2O2,), superoxide (O2.−) production rate, and the degree of lipid peroxidation were measured. Furthermore, antioxidant enzymes activities, superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and enzymes of the ascorbate–glutathione (AsA-GSH) and glutathione peroxidase (GPX) cycles were investigated. Enzymatic activity results of select genes were confirmed using real-time quantitative polymerase chain reaction (RT-qPCR). Meyerozyma caribbica pretreated with 10 μmol/mL PA (YE) was most efficient in reducing sour decay in grape berries as indicated by an 81.1 % reduction in disease incidence and 1.5 mm lesion diameter by the third day. Phytic acid treated M. caribbica showed a significant reduction in the contents of malondialdehyde (MDA), H2O2, and production rate of O2.− which was complemented by a marked rise in ROS scavenging enzymes’ activities. The H2O2 content in YE was the lowest (24.09 nmol/g FW) by the third day and O2.− production rate was about half that of CK; control (4.19 mmol/min.g FW) on the same day. The SOD, POD, and CAT activities were significantly higher in YE treated fruits (6.63, 18, and 6.64 U/mg FW, respectively) compared to the CK by the fifth day. Subsequently, AsA-GSH and GPX cycles also showed higher activity in grapes inoculated with YE compared to other treatments. Genes encoding SOD, POD, CAT, DHAR, GR, and APX were significantly overexpressed in grapes inoculated with YE compared to the CK, yeast alone, and PA treatments. These findings indicate that PA likely enhances the biocontrol efficacy of M. caribbica, leading to the induction of ROS scavenging mechanisms and subsequent mortality of A. carbonarius in table grapes. Consequently, M. caribbica pre-treated with PA holds promise as a potential biological control agent for A. carbonarius in future applications

Curcumin as a Promising Therapy for COVID-19: A Review

The ongoing COVID-19 pandemic has spurred intense research efforts to identify effective therapeutic options. Among the potential candidates, curcumin, a naturally occurring polyphenol obtained from turmeric, has gained considerable attention due to its diverse pharmacological properties. This review examines the existing literature on curcumin’s potential as a new promising treatment for COVID-19. Curcumin demonstrates antiviral effects by modulating key signaling pathways for entry and multiplication of SARS-CoV-2 in host cells. It limits viral entry in host cells as it binds and inhibits S-protein, TMPRSS2, and ADAM17 enzymes required for cytoadherence and membrane fusion. It also downregulates SARS-CoV-2 replication by preventing the release of the viral genomic RNA into the cytoplasm from virus-containing vacuoles and subsequently inhibits enzymes required for viral replication. Rennin–angiotensin–aldosterone system (RAS) dysfunction, especially increased angiotensin-converting enzyme (ACE)-Angiotensin II-AT1R axis activity, is associated with prothrombotic state, acute respiratory distress syndrome, and lung injury in COVID-19 patients. Curcumin increases soluble ACE2 cellular ACE2 activity, restores RAS normal function, and mitigates these complications. Curcumin also exerts anti-inflammatory and immunomodulatory actions. It reduces the secretion of pro-inflammatory cytokines through inhibition of toll-like receptors (TLRs), namely, TLR2, TLR4, and TLR9, and enhances the production of anti-inflammatory cytokines like interleukin-10. In addition, it prevents the progression of tissue damage and inflammation by reactive oxygen species (ROS) through ROS scavenging enzymes. Due to its antiviral, anti-inflammatory, antioxidant, and immunomodulatory properties, curcumin has emerged as an attractive candidate for combating various aspects of COVID-19 pathogenesis, such as excessive inflammation, oxidative stress, viral multiplication, and immune dysregulation. However, limited clinical evidence is currently available to support its efficacy, specifically against COVID-19. Thus, further research, including clinical trials, is warranted to evaluate curcumin’s therapeutic potential and determine its optimal dosage, formulation, and safety for COVID-19 patients. Overall, based on its favorable pharmacological properties and promising preclinical data, curcumin holds promise as a treatment for COVID-19, but its clinical utility requires further exploration.

Melatonin Mitigates Water Deficit Stress in Cenchrus alopecuroides (L.) Thunb through Up-Regulating Gene Expression Related to the Photosynthetic Rate, Flavonoid Synthesis, and the Assimilatory Sulfate Reduction Pathway.

Melatonin can improve plant adaptability to water deficit stress by regulating the biosynthesis of flavonoids and improving the reactive oxygen species-scavenging enzyme system. However, it remains unclear whether melatonin mitigates the effects and causes of water deficit stress in Cenchrus alopecuroides. We conducted a PEG-simulated water stress pot experiment to determine whether and how exogenous melatonin alleviates water deficit in C. alopecuroides. The experiment was divided into four treatments: (1) normal watering (Control), (2) 40% PEG-6000 treatment (D), (3) 100 μmol·L-1 melatonin treatment (MT), and (4) both melatonin and PEG-6000 treatment (DMT). The results showed that melatonin can alleviate water deficit in C. alopecuroides by effectively inhibiting plant chlorophyll degradation and MDA accumulation while increasing antioxidant enzyme activities and photosynthetic rates under water deficit stress. The transcriptome results indicated that melatonin regulates the expression of genes with the biosynthesis pathway of flavonoids (by increasing the expression of PAL, 4CL, HCT, and CHS), photosynthesis-antenna proteins (by increasing the expression of LHC), and sulfur metabolism (the expression of PAPSS and CysC is up-regulated in the assimilatory sulfate reduction pathway), while up-regulating the transcription factors (AP2/ERF-ERF-, C2H2-, WRKY-, Tify-, bHLH-, NAC-, and MYB-related). These findings revealed the possible causes by which melatonin mitigates water deficit stress in C. alopecuroides, which provided novel insights into the role of melatonin in water deficit stress.

Effects of cold plasma treatment on reactive oxygen metabolism and storage quality of Brassica chinensis

Cold plasma (CP) is a promising non-thermal food preservation technique, which is used for the preservation of fruits and vegetables. In this study, we aimed to determine the effects of CP treatment on the reactive oxygen metabolism, antimicrobial properties, and antioxidant activities in Brassica chinensis during storage at 4 ± 1 °C for 10 days. Postharvest Brassica chinensis were treated with CP for 6 min (CP-6), 9 min (CP-9), and 12 min (CP-12). To maintain a balance between microbial spoilage and storage quality, CP-9 was considered to be the optimal treatment time. Compared with the control group, Brassica chinensis in the CP-9 group showed a 12.7% increased antioxidant capacity, a reduction of the microbial count by 0.7 lg CFU g−1, and a delay in weight loss and soluble solid content reduction. CP inhibited the generation of superoxide anions, improved the activity of reactive oxygen species scavenging enzymes, and delayed the decrease in total antioxidant capacity. Altogether, CP treatment can be considered an advantageous approach to improve the safety and quality of fresh vegetables.

ROS Detection and Quantification in Plant-Nematode Interactions.

Reactive oxygen species (ROS) accumulation is one of the earliest hallmarks upon successful pathogen recognition in plants. H2O2 is considered the most important ROS in plant defense considering its relatively high stability and capacity to cross long distances in the plant. However, ROS also play roles in cell development and could hence facilitate nematode feeding site development. Several methods to analyze the cellular redox state exist, among which ROS detection and quantification and the evaluation of ROS scavenging enzyme activity (peroxidase activity, catalase activity, etc.). Here, we describe DAB staining, which is used to detect and localize ROS in planta upon an external trigger. Furthermore, ROS quantification using the FOX assay is described. Both methods have been used extensively in research and yield repeatable results in various plants.

Genome-wide identification of RNA recognition motif (RRM1) in Brassica rapa and functional analysis of RNA-binding protein (BrRBP) under low-temperature stress

BackgroundThe RNA recognition motif (RRM) is primarily engaged in the processing of mRNA and rRNA following gene transcription as well as the regulation of RNA transport; it is critical in preserving RNA stability.ResultsIn this study, we identified 102 members of the RRM1 gene family in Brassica rapa, which were dispersed across 10 chromosomes with the ninth chromosome being the most extensively distributed. The RRM1 gene family members of Brassica rapa and Arabidopsis thaliana were grouped into 14 subclades (I–XIV) using phylogenetic analysis. Moreover, the results of transcriptome analysis and RT-qPCR indicated that the expression of Brapa05T000840 was upregulated in the cultivars ‘Longyou 7’ and ‘Longyou 99’ following exposure to cold stress at a temperature of 4 °C for 24 h. The levels of expression in the leaves and growth cones of the ‘Longyou 7’ variety were found to be significantly higher than those observed in the ‘Longyou 99’ variety under conditions of low temperature and NaCl stress. It illustrates the involvement of the RRM1 gene in the physiological response to both low temperature and salt stress. In addition, it was observed that the survival rate of transgenic BrRBP (Brapa05T000840) Arabidopsis thaliana plants was notably higher compared to that of wild-type plants when subjected to varying durations of low temperature treatment. Furthermore, the expression of the BrRBP gene in transgenic plants exhibited an upward trend as the duration of low temperature treatment increased, reaching its peak at 24 h. The in-vivo enzymatic activity of reactive oxygen species-scavenging enzymes were found to be significantly elevated in comparison to wild-type plants, suggesting that the BrRBP gene may enhance the cold tolerance of Arabidopsis thaliana.ConclusionsThis study offers a significant foundation for comprehending the regulation mechanism of the RRM1 gene family in winter Brassica rapa subjected to cold stress, as well as for finding key genes associated with cold resistance.

Abstract B022: Effects of RSO-021 on cytokine profiles of malignant pleural effusions from patients enrolled in the MITOPE phase 1/2 clinical trial

Abstract Tumor cells generate increased levels of reactive oxygen species (ROS) that promote growth and survival through activation of redox dependent signaling pathways. To survive under increased ROS levels tumor cells, including mesotheliomas, must increase antioxidant expression, activity and reliance on critical ROS scavenging pathways, including the mitochondrial peroxiredoxin 3 (PRX3) system. PRX3 is a critical ROS scavenging enzyme localized to the mitochondrial matrix. Our group has characterized the mechanism of action of the PRX3 inhibitor and pro-oxidant therapeutic thiostrepton (TS). TS covalently crosslinks the peroxidatic and resolving cysteines of PRX3, leading to increased levels of mitochondrial ROS, resulting in tumor cell death. TS is the active pharmaceutical ingredient of RSO-021, a new covalent inhibitor of PRX3 currently being tested in the MITOPE phase 1/2 clinical trial in patients with malignant pleural effusion (MPE) due to advanced solid tumors or mesothelioma (NCT05278975). RSO-021 is administered locally to patients through an indwelling pleural catheter (IPC) weekly. TS targets PRX3 in both adherent tumor cells and in non-adherent immune cell populations in a dose-dependent manner when tested ex vivo. Using MPE drainages from patients enrolled in the MITOPE trial we evaluated cytokine levels pre and 24 and 168 - hours (7 days) post initial administration of RSO-021. Cytokines were measured in decellularized MPE drainages using Human Cytokine ELISA plates manufactured by Signosis, Inc.Adiponectin and ICACM-1 where elevated in all patients pre and post treatment. Cytokines found to be increased in patients treated with RSO-021 included IL-1B, IL-8, and Resistin. Cytokines found to be decreased following treatment included IL-13, IL-31, and IP-10. Together, these data provide the first evidence for the molecular activity of TS in patients and the resulting changes to cytokine profiles. The downstream effects of these changes to cytokine levels on cellular composition and patient responses is an ongoing focus of our research teams. Citation Format: Terri Messier, Molly Scotland, George Naumov, Sean Dulloo, Dean Fennell, Brian Cunniff. Effects of RSO-021 on cytokine profiles of malignant pleural effusions from patients enrolled in the MITOPE phase 1/2 clinical trial [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B022.

A LlWRKY33-LlHSFA4-LlCAT2 module confers resistance to Botrytis cinerea in lily.

Gray mold caused by Botrytis cinerea is one of the major threats in lily production. However, limited information is available about the underlying defense mechanism against B. cinerea in lily. Here, we characterized a nuclear-localized class A heat stress transcription factor (HSF)-LlHSFA4 from lily (Lilium longiflorum), which positively regulated the response to B. cinerea infection. LlHSFA4 transcript and its promoter activity were increased by B. cinerea infection in lily, indicating its involvement in the response to B. cinerea. Virus-induced gene silencing (VIGS) of LlHSFA4 impaired the resistance of lily to B. cinerea. Consistent with its role in lily, overexpression of LlHSFA4 in Arabidopsis (Arabidopsis thaliana) enhanced the resistance of transgenic Arabidopsis to B. cinerea infection. Further analysis showed that LlWRKY33 directly activated LlHSFA4 expression. We also found that both LlHSFA4 and LlWRKY33 positively regulated plant response to B. cinerea through reducing cell death and H2O2 accumulation and activating the expression of the reactive oxygen species (ROS) scavenging enzyme gene LlCAT2 (Catalase 2) by binding its prompter, which might contribute to reducing H2O2 accumulation in the infected area. Taken together, our data suggested that there may be a LlWRKY33-LlHSFA4-LlCAT2 regulatory module which confers B. cinerea resistance via reducing cell death and the ROS accumulation.

Characterization of a Bacillus velezensis strain as a potential biocontrol agent against soft rot of eggplant fruits

Postharvest soft rot of eggplant fruits caused by Pectobacterium carotovorum is a bacterial disease with a high disease incidence and produces substantial economic losses. This study aimed to control postharvest soft rot of eggplant fruits by Bacillus velezensis and investigate the possible control mechanisms based on the effects of B. velezensis on P. carotovorum subsp. carotovorum (Pcc) and eggplant fruits, respectively. B. velezensis effectively controlled postharvest soft rot of eggplant fruits and directly inhibited Pcc growth in vitro. The volatile metabolites produced by B. velezensis showed no inhibition on Pcc. Whereas the cell-free filtrate of B. velezensis significantly inhibited the growth of Pcc in vitro and in vivo. Notably, methanol-soluble precipitates obtained from cell-free filtrate showed significant inhibition on Pcc, and the primary inhibitory substances were identified as surfactin isoforms. Besides, iturin and fengycin isoforms with much lower relative abundance were also detected in the methanol-soluble precipitates. Furthermore, B. velezensis enhanced the activities of reactive oxygen species (ROS) scavenging enzymes in eggplant fruits that alleviated ROS and oxidative damage; thereby, B. velezensis enhanced the fruits' disease resistance.

Overexpression of a C3HC4‐type E3‐ubiquitin ligase contributes to salinity tolerance by modulating Na+ homeostasis in rice

AbstractSoil salinity has a negative effect on crop yield. Therefore, plants have evolved many strategies to overcome decreases in yield under saline conditions. Among these, E3‐ubiquitin ligase regulates salt tolerance. We characterized Oryza sativa Really Interesting New Gene (RING) Finger C3HC4‐type E3 ligase (OsRFPHC‐4), which plays a positive role in improving salt tolerance. The expression of OsRFPHC‐4 was downregulated by high NaCl concentrations and induced by abscisic acid (ABA) treatment. GFP‐fused OsRFPHC‐4 was localized to the plasma membrane of rice protoplasts. OsRFPHC‐4 encodes a cellular protein with a C3HC4‐RING domain with E3 ligase activity. However, its variant OsRFPHC‐4C161A does not possess this activity. OsRFPHC‐4‐overexpressing plants showed enhanced salt tolerance due to low accumulation of Na+ in both roots and leaves, low Na+ transport in the xylem sap, high accumulation of proline and soluble sugars, high activity of reactive oxygen species (ROS) scavenging enzymes, and differential regulation of Na+/K+ transporter expression compared to wild‐type (WT) and osrfphc‐4 plants. In addition, OsRFPHC‐4‐overexpressing plants showed higher ABA sensitivity under exogenous ABA treatment than WT and osrfphc‐4 plants. Overall, these results suggest that OsRFPHC‐4 contributes to the improvement of salt tolerance and Na+/K+ homeostasis via the regulation of changes in Na+/K+ transporters.

Tussilagone inhibits osteoclastogenesis by modulating mitochondrial function and ROS production involved Nrf2 activation

Reactive Oxygen Species (ROS) play an essential role in the pathogenesis of osteoporosis mainly characterized by excessive osteoclasts (OCs) activity. OCs are rich in mitochondria for energy support, which is a major source of total ROS. Tussilagone (TSG), a natural Sesquiterpenes from the flower of Tussilago farfara, has plentiful beneficial pharmacological characteristics with anti-inflammatory and anti-oxidative activity, but its effects and mechanism in osteopathology are still unclear. In our study, we investigated the regulation of ROS generated from the mitochondria in OCs. We found that TSG inhibited OCs differentiation and bone resorption without any cytotoxicity. Mechanistically, TSG reduced RANKL-mediated total ROS level by down-regulating intracellular ROS production and mitochondrial function, leading to the suppression of NFATc1 transcription. We also found that nuclear factor erythroid 2-related factor 2 (Nrf2) could enhance ROS scavenging enzymes in response to RANKL-induced oxidative stress. Furthermore, TSG up-regulated the expression of Nrf2 by inhibiting its proteosomal degradation. Interestingly, Nrf2 deficiency reversed the suppressive effect of TSG on mitochondrial activity and ROS signaling in OCs. Consistent with this finding, TSG attenuated post-ovariectomy (OVX)- and lipopolysaccharide (LPS) induced bone loss by ameliorating osteoclastogenesis. Taken together, TSG has an anti-bone resorptive effect by modulating mitochondrial function and ROS production involved Nrf2 activation.

Combination of curcumin and catalase protects against chondrocyte injury and knee osteoarthritis progression by suppressing oxidative stress

Knee Osteoarthritis (KOA) is an age-related progressive degenerative joint disease, which is featured with pain, joint deformity, and disability. Accumulating evidence indicated oxidative stress plays a crucial role in the occurrence and development of KOA. Curcumin is a polyphenolic compound with significant antioxidant activity among various diseases while catalase (CAT) is an enzyme degrading hydrogen peroxide in treating oxidative diseases. We previously showed that the expression of CAT was low in cartilage. However, the combination of curcumin and CAT in KOA is still elusive. In this study, we demonstrated that the combination of curcumin and CAT has the potential to inhibit the IL1β-induced chondrocyte apoptosis without cytotoxicity in vitro. Mechanistically, we found that the synergistic application curcumin and CAT not only promotes curcumin's regulation of the NRF2/HO-1 signaling pathway to enhance antioxidant enzyme expression to remove superoxide radicals, but also CAT can further remove downstream hydrogen peroxide which enhances the ability to scavenge reactive oxygen species (ROS). In vivo, studies revealed that combination of curcumin and catalase could better inhibit oxidative stress-induced chondrocyte injury by promoting the expression of ROS scavenging enzymes. In sum, the combination of curcumin and catalase can be used to treat KOA. Thus, combination of curcumin and catalase may act as a novel therapeutic agent to manage KOA and our research gives a rationale for their combined use in the therapeutic of KOA.

Effect of tamoxifen treatment on catalase (CAT) and superoxide dismutase (SOD) expression and localization in the hen oviduct

The imbalance between free reactive oxygen species (ROS) generation and removal (e.g., by antioxidative enzymes) leads to the damage of important biomolecules and cells. Earlier studies in hens showed that treatment with tamoxifen (TMX; estrogen receptor inhibitor) modulates oxidative stress and causes the reproductive system regression realized by cell apoptosis. The aim of the present study was, therefore, to examine the expression and immunolocalization of the key enzymatic antioxidants, i.e. catalase (CAT) and superoxide dismutase (SOD), in the chicken oviduct following TMX treatment. Laying hens were treated daily with TMX until a pause in egg-laying occurred and then euthanized on day 8 of the experiment. Quantitative real-time PCR and western blot analyses showed the presence of CAT and SOD transcripts and proteins, respectively, in all oviductal segments, i.e., the infundibulum, magnum, isthmus, shell gland and vagina. In control hens (laying), the mRNA expression of CAT was the highest in the shell gland, lower in the isthmus and the lowest in other oviductal parts, whereas protein expression was the highest in the magnum, lower in the isthmus and the lowest in other segments. The SOD transcript and protein abundances only were lower in the magnum than in other segments. Immunoreactive CAT and SOD products were localized in all layers of the oviductal wall, but the intensity of staining depended on the cell type. TMX treatment affected CAT and SOD expression and the effect of TMX depended on gene, protein, cell type and oviductal part. Generally, CAT expression was elevated, while SOD expression was decreased under TMX treatment. These results point to the importance of CAT and SOD in the maintenance of proper oviduct health and function. Changes in ROS scavenging enzymes after estrogen receptor blockage indicate the significance of estrogen in the regulation of oxidative status in the avian oviduct.

Computational and experimental analysis of foxtail millet under salt stress and selenium supplementation.

Salinity stress is a major threat to crop growth and productivity. Millets are stress-tolerant crops that can withstand the environmental constraints. Foxtail millet is widely recognized as a drought and salinity-tolerant crop owing to its efficient ROS scavenging mechanism. Ascorbate peroxidase (APX) is one of the reactive oxygen species (ROS) scavenging enzymes that leads to hydrogen peroxide (H2O2) detoxification and stabilization of the internal biochemical state of the cell under stress. This inherent capacity of the APX enzyme can further be enhanced by the application of an external mitigant. This study focuses on the impact of salt (NaCl) and selenium (Se) application on the APX enzyme activity of foxtail millet using in silico and in-vitro techniques and mRNA expression studies. The NaCl was applied in the concentrations, i.e., 150mM and 200mM, while the Se was applied in 1μM, 5μM, and 10μM concentrations. The in silico studies involved three-dimensional structure modeling and molecular docking. The in vitro studies comprised the morphological and biochemical parameters, alongside mRNA expression studies in foxtail millet under NaCl stress and Se applications. The in silico studies revealed that the APX enzyme showed better interaction with Se as compared to NaCl, thus suggesting the enzyme-modulating role of Se. The morphological and biochemical analysis indicated that Se alleviated the NaCl (150mM and 200mM) and induced symptoms at 1µM as compared to 5 and 10µM by enhancing the morphological parameters, upregulating the gene expression and enzyme activity of APX, and ultimately reducing the H2O2 content significantly. The transcriptomic studies confirmed the upregulation of chloroplastic APX in response to salt stress and selenium supplementation. Hence, it can be concluded that Se as a mitigant at lower concentrations can alleviate NaCl stress in foxtail millet.

Flagellin, a plant-defense-activating protein identified from Xanthomonas axonopodis pv. Dieffenbachiae invokes defense response in tobacco

BackgroundSecretome analysis is a valuable tool to study host-pathogen protein interactions and to identify new proteins that are important for plant health. Microbial signatures elicit defense responses in plants, and by that, the plant immune system gets triggered prior to pathogen infection. Functional properties of secretory proteins from Xanthomonas axonopodis pv. dieffenbachiae (Xad1) involved in priming plant immunity was evaluated.ResultsIn this study, the secretome of Xad1 was analyzed under host plant extract-induced conditions, and mass spectroscopic analysis of differentially expressed protein was identified as plant-defense-activating protein viz., flagellin C (FliC). The flagellin and Flg22 peptides both elicited hypersensitive reaction (HR) in non-host tobacco, activated reactive oxygen species (ROS) scavenging enzymes, and increased pathogenesis-related (PR) gene expression viz., NPR1, PR1, and down-regulation of PR2 (β-1,3-glucanase). Protein docking studies revealed the Flg22 epitope of Xad1, a 22 amino acid peptide region in FliC that recognizes plant receptor FLS2 to initiate downstream defense signaling.ConclusionThe flagellin or the Flg22 peptide from Xad1 was efficient in eliciting an HR in tobacco via salicylic acid (SA)-mediated defense signaling that subsequently triggers systemic immune response epigenetically. The insights from this study can be used for the development of bio-based products (small PAMPs) for plant immunity and health.