ROS Reduction Research Articles

Protection of Qingfei Xieding prescription from idiopathic pulmonary fibrosis by regulating renin-angiotensin and ferroptosis in MLE-12 cells.

Idiopathic pulmonary fibrosis (IPF) is a lifelong lung disease, but there is no specific drug for treatment. Qingfei Xieding prescription (QF) is active in the treatment of lung diseases. More comprehensive mechanisms over how QF exhibits anti-pulmonary fibrosis need to be elucidated. TGF-β was used to construct a pulmonary fibrosis cell model in vitro. Bleomycin was applied to induce a lung tissue fibrosis model in mice in vivo. Flow cytometry was used to detect cellular ROS and lipid oxidation levels. Cell substructure was observed by Transmission Electron Microscopy. ELISA was used to determine the levels of inflammatory factors. HE staining, Masson staining and immunohistochemistry were performed to evaluate the degree of fibrosis. Western Blot assay was used to determine the protein expressions of different molecules. In TGF-β-exposed lung epithelial MLE-12 cell model, α-SMA and Collagen I were significantly elevated and cell viability was reduced. QF treatment restored the cell viability decreased by exogenous TGF-β. Ferroptosis inducer Erastin administration could reverse the beneficial effects such as lipid oxidation and ROS reduction caused by QF treatment. QF was proven to inhibit ferroptosis and alleviated the process of IPF by activating ACE2 signal axis. In bleomycin induced IPF mice model, QF altered lung coefficient, body weight and the expression of inflammatory factors, which were prevented by ferroptosis activator Erastin. QF was demonstrated to affect the ACE2-ERK signaling axis in vivo. QF alleviated idiopathic pulmonary fibrosis by regulating renin-angiotensin through blocking ferroptosis. This research offers evidence for the potentiality of QF in clinical application for IPF therapy.

Interleukin-17D accelerates atherosclerosis through promoting endothelial cells ferroptosis via CD93/miR-181a-5p/SLC7A11 signaling

IL-17D has been found to induce inflammatory cytokines in endothelial cells, but its exact role in atherosclerosis (AS) is unclear. This study aims to explore IL-17D’ function in AS development. The expression of IL-17D was examined in AS patients and mice, and its clinical significance was evaluated in patients with acute coronary syndrome (ACS). Apolipoprotein E and IL-17D deficient mice (ApoE−/−IL-17D−/−) were generated for this study. The inflammation response and ferroptosis status in vascular endothelial cells were assessed following IL-17D treatment. Flow cytometry was used to identify the functional receptor of IL-17D. Additionally, RNA-seq was utilized to analyze the miRNA expression profiles induced by IL-17D. Plasma levels of IL-17D were elevated in both AS patients and mice, and were correlated with an increased incidence of major adverse cardiovascular events (MACEs). ApoE−/−IL-17D−/− mice displayed reduced inflammation and fewer atherosclerotic lesions. Treatment with IL-17D resulted in elevated levels of IL-6, IL-8, and ROS, as well as impaired cell viability and GSH production in endothelial cells. Ferroptosis inhibitor (Fer-1) suppressed the proinflammatory effects by IL-17D. Furthermore, CD93 was identified as the functional receptor for IL-17D in endothelial cells. The inhibition of miR-181a-5p led to a significant increase in cell viability and GSH levels, alongside a reduction in ROS and IL-6/IL-8 levels, while the suppression of SLC7A11 abolished these effects. Our findings suggest that IL-17D promotes endothelial inflammation by causing ferroptosis via CD93/miR-181a-5p/SLC7A11 signaling pathway. These insights advance our understanding of the pathophysiology of AS and identify a potential target for therapeutic intervention.

Ginsenoside Rg3 activates the immune function of CD8+ T cells via circFOXP1-miR-4477a-PD-L1 axis to induce ferroptosis in gallbladder cancer.

Gallbladder cancer (GBC) is the most common and leading cause of cancer-associated mortality among biliary tract carcinomas worldwide and there is no specific drug for treatment. Activation of CD8+ T cell immune activity is one of the strategies to improve GBC treatment. This study is aimed to investigate the role of Ginsenoside Rg3 on CD8+ T cell activation and pathogenesis of GBC. In GBC cells, Rg3 administration led to the significant reduction of circFOXP1 and PD-L1 as measured by Quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting. Mechanistically, circFOXP1 acted as the sponge of miR-4477a to regulate PD-L1 expression as demonstrated by RNA pull-down assay and dual luciferase reporter assay. Rg3 treatment enhanced the activity of CD8+ T cells by inhibiting thecircFOXP1/miR-4477a/PD-L1 signaling axis. Besides, Rg3 administration induced lipid oxidation and ROS reduction as detected by Flow cytometry, resulting in ferroptosis via theinactivation of circFOXP1/miR-4477a/PD-L1 axis. Ferroptosis inhibitor Fer-1 administration could reverse the beneficial effects caused by Rg3 treatment while ferroptosis inducer Erastin treatment enhanced the effects. Moreover, Rg3 gavage alleviated tumor growth and elevated ferroptosis and apoptosis in tumor tissues, which were prevented by PD-L1 overexpression. Furthermore, Rg3 was demonstrated to activate the function of CD8+ T cells via regulating thecircFOXP1-miR-4477a-PD-L1 signaling axis in vivo. Rg3 inactivated thecircFOXP1-miR-4477a-PD-L1 signaling axis to activate the immune function of CD8+ T cells, thereby inducing ferroptosis and apoptosis in GBC cells. This research recognizes the mechanism of Rg3-mediated anti-cancer effect and offers evidence for the potentiality of Rg3 in clinical application for GBC therapy.

HMGCR Inhibitor Restores Mitochondrial Dynamics by Regulating Signaling Cascades in a Rodent Alzheimer's Disease Model.

Atorvastatin an HMGCR inhibitor may play a role in enhancing spatial and long-term memory and combating anxious behavior deficits induced by Aβ1-42. Behavioral deficit studies, immunoblotting for the antioxidant/apoptotic protein expression, flow cytometry (FACS) for mitochondrial ROS, membrane potential (▲ψm), and histopathological alterations were performed against Aβ1-42 toxicity. Aβ1-42 was infused directly into the brain through i.c.v for the establishment of the AD model. Atorvastatin (ATOR) was administered orally and was used to treat AD in adult male Wistar rats aged between 200 and 250g. We confirmed that ATOR administration significantly attenuates the Aβ1-42-induced cognitive decline targeted mitochondrial-mediated age-dependent disease progression. Nrf2 stabilizes to interact SOD2 antioxidant enzyme, allowing transcriptional activity by the steep increase in ▲ψm and a reduction in ROS by activating mitochondrial superoxide scavenger and Nrf2-dependent pathway. These findings confirmed that ATOR has the potential efficacy to modulate the interference in cognitive decline induced by Aβ1-42.

Evaluating sacha inchi (Plukenetia volubilis) oil stability and physicochemical properties: A comparison between conventional extraction and supercritical fluids

This study aimed to compare the effects of two extraction techniques (conventional n-hexane and supercritical CO2) on the oil extraction yields, fatty acids profile, anti-hyaluronidase activity, oxidative stability, and in vitro bioactivities of oils from Sacha Inchi (Plukenetia volubilis). Higher oil extraction yield (99 %) was achieved using the SC-CO2, although similar fatty acids profiles were depicted between both treatments (p < 0.05). The SC-CO2 oil presented higher anti-hyaluronidase (31 %) activity, but lower oxidative stability (5.05 h) compared to the solvent extraction (10 %, and 5.3 h, respectively). In vitro assays further revealed that the best human normal colon cells (FHC) cell viability (100 %), anti-inflammatory (50 % lower NO production), and antioxidant (20 % ROS reduction) activities were consistently observed in both extraction treatments at concentrations of 50 μg/mL and higher. These findings highlight the potential of supercritical CO2 extraction in yielding Sacha Inchi oil with enhanced bioactive properties without the disadvantages of the use of organic solvents extraction.

The ER Stress Induced in Human Neuroblastoma Cells Can Be Reverted by Lumacaftor, a CFTR Corrector.

Most neurodegenerative diseases share a common etiopathogenesis, the accumulation of protein aggregates. An imbalance in homeostasis brought on by the buildup of misfolded proteins within the endoplasmic reticulum (ER) results in ER stress in the cell. Three distinct proteins found in the ER membrane-IRE1α, PERK, and ATF6-control the unfolded protein response (UPR), a signal transduction pathway that is triggered to restore normal physiological conditions. Buildup of misfolded proteins in ER lumen leads to a shunting of GRP78/BiP, thus triggering the UPR. PERK autophosphorylation leads to activation of ATF4, the transcription factor; finally, ATF6 activates the UPR's target genes, including GRP78/Bip. Accordingly, the UPR is a cellular reaction to an ER stress state that, if left unchecked for an extended period, results in apoptosis and irreversible damage. The identification of caspase 4, which is in the ER and is selectively activated by apoptotic stimuli caused by reticular stress, further demonstrated the connection between reticular stress and programed cell death. Moreover, oxidative stress and ER stress are linked. Oxidative stress is brought on by elevated quantities of radical oxygen species, both mitochondrial and cytosolic, that are not under the enzymatic regulation of superoxide dismutases, whose levels fall with increasing stress. Here, we evaluated the activity of Vx-809 (Lumacaftor), a drug used in cystic fibrosis, in SH-SY5Y neuronal cells, in which an ER stress condition was induced by Thapsigargin, to verify whether the drug could improve protein folding, suggesting its possible therapeutic use in proteinopathies, such as neurodegenerative diseases (NDs). Our data show that Vx-809 is involved in the significant reduction in protein produced under ER stress, particularly in the levels of Bip, ATF4, and ATF6 by Western blotting analysis, the reduction in ROS in the cytosol and mitochondria, and the reduction in the activation of the apoptotic pathway, measured by flow cytofluorimetry analysis and in restoring calcium homeostasis.

AMPK activation reduces cancer cell aggressiveness via inhibition of monoamine oxidase A (MAO-A) expression/activity

AimAMPK can be considered as an important target molecule for cancer for its unique ability to directly recognize cellular energy status. The main aim of this study is to explore the role of different AMPK activators in managing cancer cell aggressiveness and to understand the mechanistic details behind the process. Main methodsFirst, we explored the AMPK expression pattern and its significance in different subtypes of lung cancer by accessing the TCGA data sets for LUNG, LUAD and LUSC patients and then established the correlation between AMPK expression pattern and overall survival of lung cancer patients using Kaplan-Meire plot. We further carried out several cell-based assays by employing different wet lab techniques including RT-PCR, Western Blot, proliferation, migration and invasion assays to fulfil the aim of the study. Key findings•Expression of AMPK is correlated with survival and prognosis of lung cancer patients.•AMPK activators like Metformin and Phenformin downregulate A549 and HCT-116 cell proliferation and migration via repression of p38MAPK activity, subsequent augmentation of R1 repressor and corresponding downregulation of MAO-A expression/activity resulting reduction in the intracellular ROS.•SRT-1720 directly activates AMPK in LKB1 mutant A549 cells either alone or in combination with Metformin resulting regulation of cancer cell aggressiveness. SignificanceThis study identifies the importance of AMPK activators as a repurposing agent for combating lung and colon cancer cell aggressiveness. It also suggests SRT-1720 as a potent repurposing agent for cancer treatment especially in NSCLC patients where a point mutation is present in LKB1.

Adjustments of the Phytochemical Profile of Broccoli to Low and High Growing Temperatures: Implications for the Bioactivity of Its Extracts.

Climate change causes shifts in temperature patterns, and plants adapt their chemical content in order to survive. We compared the effect of low (LT) and high (HT) growing temperatures on the phytochemical content of broccoli (Brassica oleracea L. convar. botrytis (L.) Alef. var. cymosa Duch.) microgreens and the bioactivity of their extracts. Using different spectrophotometric, LC-MS/MS, GC-MS, and statistical methods, we found that LT increased the total phenolics and tannins in broccoli. The total glucosinolates were also increased by LT; however, they were decreased by HT. Soluble sugars, known osmoprotectants, were increased by both types of stress, considerably more by HT than LT, suggesting that HT causes a more intense osmotic imbalance. Both temperatures were detrimental for chlorophyll, with HT being more impactful than LT. HT increased hormone indole-3-acetic acid, implying an important role in broccoli's defense. Ferulic and sinapic acid showed a trade-off scheme: HT increased ferulic while LT increased sinapic acid. Both stresses decreased the potential of broccoli to act against H2O2 damage in mouse embryonal fibroblasts (MEF), human keratinocytes, and liver cancer cells. Among the tested cell types treated by H2O2, the most significant reduction in ROS (36.61%) was recorded in MEF cells treated with RT extracts. The potential of broccoli extracts to inhibit α-amylase increased following both temperature stresses; however, the inhibition of pancreatic lipase was increased by LT only. From the perspective of nutritional value, and based on the obtained results, we conclude that LT conditions result in more nutritious broccoli microgreens than HT.

Feedback regulation of the isoprenoid pathway by SsdTPS overexpression has the potential to enhance plant tolerance to drought stress.

In order to maintain the dynamic physiological balance, plants are compelled to adjust their energy metabolism and signal transduction to cope with the abiotic stresses caused by complex and changeable environments. The diterpenoid natural compound and secondary metabolites, sclareol, derived from Salvia sclarea, has gained significant attention owing to its economic value as a spice material and diverse physiological activities. Here, we focused on the roles and regulatory mechanisms of the sclareol diterpene synthase gene SsdTPS in the resistance of S. sclarea to abiotic stresses. Our results suggested that abiotic stresses could induce the response and upregulation of SsdTPS expression and isoprenoid pathway in S. sclarea. Ectopic expression of SsdTPS conferred drought tolerance in transgenic Arabidopsis, compared with wild-type. Overexpression of SsdTPS enhanced the transcription of ABA signal transduction synthetic regulators and induced the positive feedback upregulating key regulatory genes in the MEP pathway, thereby promoting the increase of ABA content and improving drought tolerance in transgenic plants. In addition, SsdTPS-overexpressed transgenic Arabidopsis improved the responses of stomatal regulatory genes and ROS scavenging enzyme activities and gene expression to drought stress. This promoted the stomatal closure and ROS reduction, thus enhancing water retention capacity and reducing oxidative stress damage. These findings unveil the potentially positive role of SsdTPS in orchestrating multiple regulatory mechanisms and maintaining homeostasis for improved abiotic stress resistance in S. sclarea, providing a novel insight into strategies for promoting drought resistance and cultivating highly tolerant plants.

Unveiling the potential of Butylphthalide: inhibiting osteoclastogenesis and preventing bone loss.

Osteoporosis, resulting from overactive osteoclasts and leading to elevated fracture risk, has emerged as a global public health concern due to the aging population. Therefore, inhibiting osteoclastogenesis and bone resorption function represents a crucial approach for preventing and treating osteoporosis. The purpose of this study was to examine the effects and molecular mechanisms of Butylphthalide (NBP) on the differentiation and function of osteoclasts induced by RANKL. Osteoclastogenesis was assessed through TRAP staining and bone slice assay. An animal model that underwent ovariectomy, simulating postmenopausal women's physiological characteristics, was established to investigate the impact of Butylphthalide on ovariectomy-induced bone loss. To delve deeper into the specific mechanisms, we employed Western blot, PCR, immunofluorescence, and immunohistochemical staining to detect the expression of proteins that are associated with the osteoclast signaling pathway. In this study, we found that Butylphthalide not only suppressed osteoclastogenesis and bone resorption in vitro but also significantly decreased TRAcP-positive osteoclasts and prevented bone loss in vivo. Further mechanistic experiments revealed that Butylphthalide reduces intracellular ROS in osteoclasts, inhibits the MAPK and NFATc1 signaling pathways, and downregulates the key genes and proteins of osteoclasts. This inhibits osteoclast formation and function. The reduction in ROS in osteoclasts is intricately linked to the activity of Butylphthalide-modulated antioxidant enzymes. Overall, NBP may offer a alternative treatment option with fewer side effects for skeletal diseases such as osteoporosis.

Exploring the ROS reduction strategies in chronic lupus management.

Exploring the ROS reduction strategies in chronic lupus management.

Exploring the Potential of Isalo Scorpion Cytotoxic Peptide in Enhancing Gill Barrier Function and Immunity in Grass Carp (Ctenopharyngodon idella) Infected with Flavobacterium columnare.

The objective of this research was to investigate how dietary antimicrobial peptides (AMP), namely, Isalo scorpion cytotoxic peptide (IsCT), affect the gill physical barrier function and immune function of grass carp challenged with Flavobacterium columnare (F. columnare). Five hundred forty grass carp were randomly allocated to six groups and fed to varying levels of IsCT in the diet (0, 0.6, 1.2, 1.8, 2.4, and 3.0 mg/kg diet) for a duration of 60 days. Afterward, the grass carps faced a challenge from F. columnare. The results revealed that the use of optimal IsCT dramatically mitigated gill damage in grass carp that were infected with F. columnare. Additionally, IsCT exhibited a notable enhancement in gill antioxidant capabilities, as evidenced by a significant reduction in ROS, MDA, and PC levels, an elevation in antioxidant enzyme activities, and an upregulation of antioxidant-related genes and Nrf2 mRNA levels. Conversely, the expression of Keap1a and Keap1b mRNA was decreased. Besides, IsCT exhibited its capability to inhibit apoptosis via downregulating the mRNA levels of caspase-2, caspase-3, caspase-7, caspase-8, caspase-9, Apaf1, Fasl, Bax, and JNK while concurrently increasing the mRNA levels of Bcl-2, Mcl-1, and IAP in fish gills. Additionally, IsCT promoted the integrity of tight junction barrier by increasing the gene expression of claudin-b, claudin-c, claudin-3c, ZO-1, ZO-2b, occludin, and JAM while suppressing MLCK signaling. Additionally, optimal dietary IsCT improved antibacterial ability, as evidenced by heightened LZ, ACP activities, and elevated levels of C3, C4, and IgM. Additionally, there was an upregulation in β-defensin-1, LEAP-2A, LEAP-2B, hepcidin, and mucin-2 mRNA expression in the gills. Simultaneously, the inclusion of optimal dietary IsCT in the diet resulted in improved gill immunity barriers through the reduction of proinflammatory cytokine mRNA levels and the increase in the expression of anti-inflammatory cytokine mRNA levels. This was partly facilitated by the IκBa/NF-κB p65 signaling pathway and TOR/S6K1 signaling pathways in the gills of grass carp. Therefore, supplementing the diet with IsCT has potential advantages in enhancing gill health by improved physical barriers and immunity in grass carp. Based on LZ activity and against lipid peroxidation, optimum IsCT concentrations in on-growing grass carp (136.88 ± 0.72 g) were found to be 1.68 and 1.54 mg/kg diet, respectively.

Atrial natriuretic peptide protects against gut barrier injury through PLC-γ1/ROS feedback loop in rats following traumatic hemorrhagic shock

Introduction The mechanisms underlying the protective effects of atrial natriuretic peptide (ANP) on the gut barrier during traumatic hemorrhagic shock (THS) remain elusive. This study aimed to explore the potential role of ANP in safeguarding against gut barrier dysfunction after THS, focusing on the PLC-γ1/ROS feedback loop. Methods In our THS rat model, we randomly allocated male Sprague-Dawley rats to receive intravenous ANP with or without a concurrent NADPH oxidase/p38 MAPK inhibitor during the shock phase. After 24 h, we assessed circulatory and jejunal ANP, ROS, intestinal tight junction proteins, and apoptosis to evaluate the effects of ANP on the gut barrier and its interplay with intestinal ANP and ROS. Rat small intestinal epithelial cells (IECs) were also treated with ANP and subjected to hypoxia/re-oxygenation injury, with or without PI3K/PLC inhibition, to elucidate the relationship between ANP/ROS signaling and PLC-γ1. Furthermore, we modulated PLC-γ1 expression in these IECs to examine its impact on ROS and ANP production. Results Intravenous ANP administration at 0.025 μg/kg/min during THS significantly increased intestinal ANP and ROS levels at 24 h. ANP treatment enhanced the expression of intestinal tight junction proteins and reduced IEC apoptosis. Inhibition of circulatory ROS diminished intestinal ANP levels, while suppression of circulatory ANP led to a reduction in intestinal ROS. Decreasing PLC-γ1 expression in hypoxia/re-oxygenation-treated IECs resulted in lower ROS and ANP levels, whereas augmenting PLC-γ1 expression did not alter these levels. Additionally, PI3K inhibition markedly decreased PLC-γ1 expression in these cells. Conclusion ANP-induced protection of the intestinal barrier in THS is mediated by an intrinsic PLC-γ1/ROS positive feedback loop. ANP preserves gut barrier integrity and reduces IEC apoptosis through this mechanism. Further studies are warranted to investigate the interaction between IECs and other cellular components within the PLC-γ1/ROS loop.

Mitochondria-targeted docetaxel loaded DQAsomes for the treatment of multi-drug resistant breast tumor: In vitro and in vivo studies

Multidrug resistance (MDR) has become a major obstacle in the successful chemotherapy of breast cancer which is the leading cause of death in women. In our recent work, we successfully demonstrated the remarkable anticancer potential of docetaxel-loaded DQAsomes (DOC-DQAsomes) against MDR-resistant MDA-MB-231 and normal MCF-7 breast cancer cell lines. However, the in vitro performance of any drug delivery system does not guarantee similar behaviour in vivo. Thus the present work was planned to assess the in vivo performance of prepared DOC-DQAsomes and to generate additional evidence for their anticancer activity. In vitro, cell uptake studies, ROS assay and mitochondrial targeting assays were carried out on DQAsomes along with in vivo antitumor efficacy in the MDR-resistant MDA-MB-231 tumor-induced SCID mice. Rhodamine-labelled DQAsomes were readily taken up by MDA-MB-231 cells in a time-dependent manner. Further DOC-DQAsomes showed an increment in ROS generation and reduction in mitochondrial membrane potential which revealed their anticancer activity. Additionally, DOC-DQAsomes demonstrated remarkable in vivo antitumor activity in tumour-bearing mice. The anticancer potential of DOC-DQAsomes was attributed to their ability to reduce mitochondrial membrane potential, generation of ROS and in turn release cytochrome C from mitochondria, leading to enhancement in apoptosis of MDA-MB-231 cells. Moreover, the treatment of DOC-DQAsomes did not affect the survival rate of xenografts which is the major drawback of many anticancer formulations. Thus DOC-DQAsomes could be the choice of treatment in MDR-resistant breast cancer demonstrating activity through the mitochondria signaling pathway.

Abstract C036: Investigating the phenotypic changes in in vitro and in vivo cancer models expressing isocitrate dehydrogenase 1 (IDH1) mutations R132H and R132Q

Abstract According to the National Cancer Institute, there were 25,050 new brain cancer cases in 2022, representing 1.3% of all cancer incidents in the United States. In the same year, the estimated death was 18,280 cases, representing 3.0% of all cancer deaths. Relative survival statistics between 2017 and 2018 showed that only 32.5% of patients would survive five years or more. Disparities in cancer incidence and outcomes, including brain cancers, can be due to biological, socioeconomic, and care access factors that often have roots in structural racism. Research shows lower three years survival rates of glioblastoma in the African American population by 28.8% compared to the Asian Pacific Islanders at 36.6% and Hispanic at 32.3%. Mutant isocitrate dehydrogenase 1 (IDH1) can lead to brain cancer, including &amp;gt; 80% of lower-grade gliomas and secondary glioblastomas. Mutant IDH1 catalyzes the NADPH-dependent conversion of α-ketoglutarate to D2-hydroxyglutarate (D2HG), an oncometabolite linked to many cancer types. IDH1 mutation occurs primarily at residue 132, and our lab has shown that the catalytic activity differs based on the amino acid substituted at that location. There is a lack of information describing the relation of D2HG produced to cancer cells’ phenotype severity. Understanding IDH1 mutation will clarify cancer progression, patients’ prognosis, and treatment responses. Here, we hypothesize that the toxic effect of D2HG will impair the viability of cells expressing mutant IDH1 and will cause a reduction in ROS and NADPH cellular levels. This work examines D2HG production efficiency in vivo and in vitro and the metabolic consequences of this oncometabolite. We created cell lines stably overexpressing IDH1-R132Q and IDH1-R132H to conduct this work. Our results showed higher levels of D2HG in cells expressing IDH1-R132Q compared to DH1-wild type and IDH1-R132H, supportive of our previous finding that IDH1-R132Q mutation has higher catalytic activities (higher efficiency in producing mutant IDH1). These higher D2HG levels caused cells to proliferate and migrate at lower rates. D2HG interferes with DNA demethylation by inhibiting the TET enzyme, and we observed higher levels of histone protein expression in IDH1-R132Q compared to the other modified cells. In in vivo studies, we identified elevated D2HG levels in mouse xenografts expressing IDH1-R132Q that affected tumor growth. Citation Format: Mowaffaq A.A. Adam, Mikella Robinson, An Hoang, Grace A. Wells, David A. Scott, Olga Zagnitko, Carrie House, Christal D. Sohl. Investigating the phenotypic changes in in vitro and in vivo cancer models expressing isocitrate dehydrogenase 1 (IDH1) mutations R132H and R132Q [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C036.

Simultaneous irradiation of 660 and 808 nm on gingival epithelial cells and fibroblasts induces different patterns of oxidative/antioxidative activities: What is the role of the cell type and irradiation parameters?

The aim of this study was to investigate whether simultaneous irradiation at 660 and 808 nm generates different patterns of oxidative/antioxidative activities compared to consecutive irradiation. Primary cultures of gingival keratinocytes and fibroblasts were exposed to a diose laser (660 ± 2 nm and 808 ± 2 nm, 100 mW, 0.09 cm2 spot area) using double irradiation with the two wavelengths (consecutive or simultaneous) for 6, 10, and 20 s. The two irradiation regimens did not increase cell viability in any of the experimental conditions. Lipid peroxidation was increased after consecutive irradiation in epithelial cells, which was not detected after simultaneous irradiation. After 20s of the simultaneous mode,ROS levels increased, but antioxidative balance decreased. In the fibroblasts, the two double irradiations induced ROS reduction, increase in lipid peroxidation, and improvement of antioxidative balance, mainly after the 20 s irradiation time. In conclusion, simultaneous and consecutive irradiation induced distinct oxidative stress modulation in oral epithelial cells and fibroblasts. The imbalance in the oxidative system observed after longer exposures, allied with the absence of a significant increase in the viability of the two cell types, suggests a contraindication for longer simultaneous irradiation in clinical situations that demand cellular stimulation.

Mitochondrial dysfunction in the pathogenesis of endothelial dysfunction.

Cardiovascular diseases (CVDs) are a matter of concern worldwide, and mitochondrial dysfunction is one of the major contributing factors. Vascular endothelial dysfunction has a major role in the development of atherosclerosis because of the abnormal chemokine secretion, inflammatory mediators, enhancement of LDL oxidation, cytokine elevation, and smooth muscle cell proliferation. Endothelial cells transfer oxygen from the pulmonary circulatory system to the tissue surrounding the blood vessels, and a majority of oxygen is transferred to the myocardium by endothelial cells, which utilise a small amount of oxygen to generate ATP. Free radicals of oxide are produced by mitochondria, which are responsible for cellular oxygen uptake. Increased mitochondrial ROS generation and reduction in agonist-stimulated eNOS activation and nitric oxide bioavailability were directly linked to the observed change in mitochondrial dynamics, resulting in various CVDs and endothelial dysfunction. Presently, the manuscript mainly focuses on endothelial dysfunction, providing a deep understanding of the various features of mitochondrial mechanisms that are used to modulate endothelial dysfunction. We talk about recent findings and approaches that may make it possible to detect mitochondrial dysfunction as a potential biomarker for risk assessment and diagnosis of endothelial dysfunction. In the end, we cover several targets that may reduce mitochondrial dysfunction through both direct and indirect processes and assess the impact of several different classes of drugs in the context of endothelial dysfunction.

Cellular antioxidant and cytotoxic activity of astaxanthin and ellagic acid on UV irradiated skin melanoma cells and gel formulation

Ultraviolet radiation accounts for most non-melanoma skin cancers (carcinoma of basal and squamous cells) and melanoma skin cancers. Hence, UV radiation is considered one of the most potent physical carcinogens. Radiation exposure leads to water hydrolysis and heightens free radical production. The free radicals, in turn, react with macromolecules such as proteins, DNA, RNA, and lipids and alters their structure. Antioxidants protect the cells against radiation-induced cell death and DNA damage. It was also attributed to the reduction in ROS induced by UV radiation. Both ellagic acid and astaxanthin has antioxidant property. In addition to the antioxidant property, these compounds possess inflammatory, anti-proliferative, and wound-healing properties. The study aims to evaluate the ROS scavenging potential of ellagic acid and astaxanthin in Skin melanoma (SK-Mel-28) cells and formulate a gel that can treat radiation dermatitis. The spectral characterization of ellagic acid and astaxanthin was carried out using UV–Vis spectroscopy and FTIR. The antioxidant potential of astaxanthin and ellagic acid was measured using probe 2′, 7′-dichlorofluorescein-diacetate in UV-treated skin melanoma cells. These antioxidants attenuated the intracellular ROS in UV-treated SK-Mel-28 cells, evident at 25 μg/ml and 50 μg/ml. Comparative fluorescent intensity profile analyzed using ImageJ also showed that both astaxanthin and ellagic acid exhibit high ROS scavenging potential without prooxidant capacity. The ellagic acid extract and astaxanthin decreasedSK-Mel-28 cell viability with a median inhibitory concentration IC50 of 47.23 μg/ml and 41.39 μg/ml. The ellagic acid enhanced thiobarbituric acid reactive substance level, an indicator of lipid peroxidation with a median stimulatory concentration EC50 of 53.7 μg/ml in SK-Mel-28 cells. This substantiated that ellagic acid potentiates the antitumor effect through lipid peroxidation. It was proposed that the astaxanthin-ellagic acid gel formulation produced better results in reducing UV-induced skin cancers due to positive synergism.

The Protective Role of Glutathione against Doxorubicin-Induced Cardiotoxicity in Human Cardiac Progenitor Cells.

This study investigated the protective effect of glutathione (GSH), an antioxidant drug, against doxorubicin (DOX)-induced cardiotoxicity. Human cardiac progenitor cells (hCPCs) treated with DOX (250 to 500 nM) showed increased viability and reduced ROS generation and apoptosis with GSH treatment (0.1 to 1 mM) for 24 h. In contrast to the 500 nM DOX group, pERK levels were restored in the group co-treated with GSH and suppression of ERK signaling improved hCPCs' survival. Similarly to the previous results, the reduced potency of hCPCs in the 100 nM DOX group, which did not affect cell viability, was ameliorated by co-treatment with GSH (0.1 to 1 mM). Furthermore, GSH was protected against DOX-induced cardiotoxicity in the in vivo model (DOX 20 mg/kg, GSH 100 mg/kg). These results suggest that GSH is a potential therapeutic strategy for DOX-induced cardiotoxicity, which performs its function via ROS reduction and pERK signal regulation.

Melatonin supplementation to the freezing medium enhances post-thaw sperm quality and fertility of giant grouper (Epinephelus lanceolatus)

Giant grouper (Epinephelus lanceolatus) is not only favored in the international live reef-fish trade but also a promising species for mariculture in coastal communities throughout Asia. Sperm cryopreservation contributes to the extensive utilization of artificial insemination in grouper aquaculture industry. Inevitable damage during the process of freezing can negatively affect post-thaw sperm quality and fertility, thus hindering its routine application in practice. Melatonin is an antioxidant that can protect sperm from the adverse effects of cryopreservation in a variety of species. This study was aimed to assess the effect of melatonin addition on post-thaw sperm function of giant grouper. Semen samples were cryopreserved with freezing medium containing different final concentrations of melatonin (1, 2 and 4 μM) and the control group (without melatonin). After thawing, multiple sperm functional parameters were evaluated. Generally, a positive effect on cryopreservation was observed for all doses of melatonin. Melatonin treatment at 1 and 2 μM seemed to be efficient in preventing sperm cryodamage, which yielded significantly higher total motility, progressive motility, mitochondrial membrane potential (P < 0.05) and improved velocity (VCL, VAP, VSL) compared to controls. Lower apoptotic-like changes were detected at 2 μM treatment. Samples frozen with melatonin also contributed to greater sperm viability and lesser DNA damage. Although not statistically different from control, a reduction in intracellular ROS was found under the melatonin treatments. Moreover, melatonin supplementation improved the fertility of frozen-thawed sperm, which increased significantly the hatching rate under a limited sperm/egg ratio (10,000:1). Hence, our study suggests inclusion of melatonin to semen extender can improve the post-thaw quality of cryopreserved giant grouper spermatozoa, the recommended concentration of melatonin is 1–2 μM in the cryomedium.