Levels Of Stress Markers Research Articles

Farnesol Inhibits Cell Viability and Activates Apoptosis in Glioma C6 Cells via Downregulating Phosphoinositide 3-kinase (PI3K)/Protein Kinase B (AKT)/Mammalian Target of Rapamycin (mTOR) Pathway

Background Gliomas are the most aggressive and devastating brain tumors, with a poor prognosis due to their complex etiology, diverse clinical manifestations, and the limited efficacy of existing treatment methods. Purpose The present work was conducted to understand the anticancer activities of farnesol against glioma C6 cells. Methods The effects of farnesol on in vitro free radical scavenging were evaluated at various dosages (1–100 µM). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was used to assess the influence of farnesol on the growth of glioma C6 cells and nonmalignant Vero cells. The apoptotic levels in the cells were investigated using 4′,6-diamidino-2-phenylindole (DAPI) and dual staining assays. The levels of oxidative stress markers, inflammatory biomarkers, apoptotic protein levels, and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway proteins were assessed using the respective test kits. Results The results of the free radical scavenging assays proved the antioxidant properties of farnesol. The MTT assay findings demonstrated a significant reduction in C6 cell viability following treatment with farnesol. The findings of the fluorescent assays proved the onset of apoptosis in the farnesol-treated C6 cells. Furthermore, the farnesol treatment effectively increased oxidative stress, increased apoptotic protein levels, and reduced inflammatory marker levels in the C6 glioma cells. The PI3K, AKT, and mTOR protein expression were also successfully downregulated in the C6 glioma cells by farnesol. Conclusion The current results demonstrate that farnesol effectively suppresses viability and triggers apoptosis in C6 cells. Thus, it has the capacity to be an effective anticancer agent to treat glioma. Nonetheless, additional work is still necessary in the future to determine the specific molecular mechanisms involved in farnesol-induced apoptosis in C6 cells.

Blackcurrant (Ribes nigrum L.) and Its Association with Donepezil Restore Cognitive Impairment, Suppress Oxidative Stress and Pro-inflammatory Responses, and Improve Purinergic Signaling in a Scopolamine-Induced Amnesia Model in Mice.

Purinergic signaling plays a major role in aging and neurodegenerative diseases, which are associated with memory decline. Blackcurrant (BC), an anthocyanin-rich berry, is renowned for its antioxidant and neuroprotective activities. However, evidence on the effects of BC on purinergic signaling is lacking. This study investigated the effects of BC and its association with Donepezil (DNPZ) on learning and memory, on the modulation of purinergic signaling, pro-inflammatory responses, and oxidative markers in a mouse model of cognitive impairment chronically induced by scopolamine (SCO). Animals were divided into twelve groups and treated with BC (50 or 100mg/kg), and/or DNPZ (5mg/kg), and/or SCO (1mg/kg). Results showed that SCO decreased spatial learning and memory as assessed by the Morris Water Maze test, and treatment with BC and/or DNPZ restored these effects. Furthermore, BC and/or DNPZ treatments also prevented changes in ecto-nucleoside triphosphate diphosphohydrolase (NTPDase) and adenosine deaminase (ADA) activities and restored the increased density of P2X7 and A2A receptors in synaptosomes of the cerebral cortex of SCO-induced mice. Moreover, the increased Nod-like receptor protein 3 (NLRP3) and interleukin-1β expression, and the oxidative stress markers levels were reduced by BC and/or DNPZ treatments, compared with the SCO group. Overall, BC and/or DNPZ treatments ameliorated SCO-induced cognitive decline, alleviated oxidative stress and pro-inflammatory responses, and improved purinergic signaling. These findings underscore the potential of BC, especially when in combination with DNPZ, as a therapeutic agent for the prevention of memory deficits associated with aging or neurological diseases.

Evaluation of the therapeutic effects of nebulized inhalation of hydrogen-rich water on primary blast lung injury in C57BL/6 mice.

Primary blast lung injury is a common and severe consequence of explosion events, characterized by immediate and delayed effects such as apnea and rapid shallow breathing. The overpressure generated by blasts leads to alveolar and capillary damage, resulting in ventilation-perfusion mismatch and increased intrapulmonary shunting. This reduces the effective gas exchange area, causing hypoxemia and hypercapnia. Hydrogen (H2), a small-molecular-weight, nonpolar diatomic molecule, has shown potential in treating various diseases due to its antioxidant and anti-inflammatory properties. This study evaluates the therapeutic effects of nebulized hydrogen-rich water on primary blast lung injury in C57BL/6 mice and explores the underlying mechanisms. C57BL/6 mice (n= 150), aged 6-8weeks, were randomly divided into 2 groups: the blast injury control group (n= 75) and the nebulized hydrogen-rich water treatment group (n= 75). Mice were exposed to a blast overpressure of 266±9.156 kPA and treated with either nebulized hydrogen-rich water or sterile injection water immediately postinjury. Observations were made at 6, 12, 24, and 48hours postinjury. Lung function was assessed using whole-body plethysmography, and arterial blood gases were analyzed. Lung tissue was examined histologically and biochemically for markers of inflammation and oxidative stress. The survival rates at different time points postinjury in the nebulized hydrogen-rich water treatment group were significantly higher than those in the blast injury control group (6hours postinjury: 89.3% vs 78.6%; 12hours postinjury: 81.3% vs 72%; 24hours postinjury: 81.3% vs 61.3%, P < .01). Lung function tests revealed significant improvements in tidal volume (12hours postinjury: 0.11±0.018 vs 0.08±0.016, 24hours postinjury: 0.16±0.013 vs 0.12±0.013, 48hours postinjury: 0.18±0.02 vs 0.13±0.014), respiratory rate (6hours postinjury: 235.07±12.82 vs 268.29±13.73; 12hours postinjury: 265.47±10.06 vs 342.16±16.34; 24hours postinjury: 248.20±9.28 vs 352.80±15.99; 48hours postinjury: 226.12±15.81 vs 318.18±15.81), and minute ventilation (12hours postinjury: 22.05±3.46 vs 15.93±3.68; 24hours postinjury: 27.30±2.15 vs 21.62±2.48; 48hours postinjury: 37.48±3.93 vs 28.32±2.98) in the nebulized hydrogen-rich water treatment group (P < .01). Arterial blood gas analysis indicated better oxygenation and reduced hypercapnia in the nebulized hydrogen-rich water treatment group (P < .05). Histologic examination showed reduced lung edema and hemorrhage in the nebulized hydrogen-rich water treatment group. Levels of inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor α) and oxidative stress markers (ie, malondialdehyde) were significantly lower, whereas antioxidant enzyme (total superoxide dismutase) activity was higher in the nebulized hydrogen-rich water treatment group compared with the blast injury control group (P < .01). Arterial blood gas analysis indicated better oxygenation and reduced hypercapnia in the nebulized hydrogen-rich water treatment group (P < .05). Histologic examination showed reduced lung edema and hemorrhage in the nebulized hydrogen-rich water treatment group. Levels of inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor α) and oxidative stress markers (ie, malondialdehyde) were significantly lower, whereas antioxidant enzyme (total superoxide dismutase) activity was higher in the nebulized hydrogen-rich water treatment group compared with the blast injury control group (P < .01). Hydrogen-rich water treatment significantly improves survival rates and lung function and reduces inflammation and oxidative stress in mice with primary blast lung injury. These findings suggest that hydrogen's antioxidant and anti-inflammatory properties play a crucial role in mitigating lung damage and improving respiratory function postinjury. Further long-term studies and imaging analyses are needed to confirm these findings and elucidate the molecular mechanisms involved. This study provides a theoretical basis for the clinical application of hydrogen-rich water in treating blast-induced lung injuries.

Tocilizumab alleviated lipopolysaccharide-induced acute lung injury by improving PI3K/AKT pathway.

Acute lung injury (ALI) is a severe inflammatory condition of the respiratory system, associated with high morbidity and mortality. This study investigates the therapeutic potential of tocilizumab (TZ), an IL-6 receptor inhibitor, in mitigating lipopolysaccharide (LPS)-induced ALI by modulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway. An ALI model was established using LPS induction. Lung damage was assessed by hematoxylin-eosin (H&E) staining, alongside measurements of respiratory function, including PaO2/Fio2 ratios, lung edema, airway resistance, and lung compliance. Western blotting analyzed the expression of phosphorylated PI3K and AKT (P-PI3K, P-AKT), while ELISA quantified levels of TNF-α, IL-1β, IL-6, and oxidative stress markers. Apoptosis was evaluated using the TUNEL assay, and key apoptotic proteins (Bcl-2, Bax, and caspase-3) were measured by Western blotting. The Cell Counting Kit-8 (CCK-8) assay was employed to determine cell viability. The LPS-induced model exhibited decreased P-PI3K and P-AKT levels, while TZ treatment significantly elevated these markers. TZ also reduced lung tissue damage, improved respiratory function, and decreased inflammation, oxidative stress, and apoptosis. However, co-administration with LY294002 (a PI3K inhibitor) blocked these benefits, reversing the protective effects of TZ. TZ alleviates lung injury and improves outcomes in LPS-induced ALI by enhancing the PI3K/AKT pathway.

The profile of oxidative stress markers (arachidonic and linoleic acid derivatives) in patients with benign prostatic hyperplasia in relation to metabolic syndrome.

So far, it has been proven that benign prostatic hyperplasia (BPH) is strongly associated with inflammation resulting from, i.a. the presence of infectious agent, autoimmune disease, aging process and lipid disorders associated with metabolic syndrome (MetS). We analyzed the association between serum eicosanoides (HETE, HODE, lipoxins, prostaglandin, and leucotrien) in aging man with benign prostatic hyperplasia (BPH) and healthy controls. The study involved 219 men (with BPH, n = 144; healthy controls, n = 75). We assessed the content arachidonic and linoleic acid derivatives in the serum samples of the study participants using liquid chromatography (HPLC). The levels of: RvE1 (p < 0.001); LXA4 5S,6R,15R (p = 0.001); 10S,17R-DiDHA (p < 0.001); MaR1 (p = 0.002); 9S-HODE (p < 0.05); 15S-HETE (p < 0.05); 12S-HETE (p < 0.001); 5-oxoETE (p < 0.05) and 5-HETE (p < 0.001) were significantly higher in patients with BPH than in the control group. PGE2 (p = 0.007), LTB4 (p < 0.001), and 18RS-HEPE (p < 0.001) were significantly higher in control group. We also analyzed the relationship between LXA4 5S,6R,15R serum levels of oxidative stress markers and concomitance of MetS. We noticed a relationship between levels and MetS (F1216 = 6.114965, p = 0.01). Our research results suggest that pro-inflammatory mediators and suppressors of inflammation are involved in the development of BPH, but their exact contribution has yet to be investigated.

Depressive symptoms in older adults are associated with changes in stress-related markers, functional connectivity and brain volume

BackgroundSubclinical depressive symptoms increase the risk of developing Alzheimer’s disease (AD). The neurobiological mechanisms underlying this link may involve stress system dysfunction, notably related to the hippocampus which is particularly sensitive to AD. We aimed to investigate the links between blood stress markers and changes in brain regions involved in the stress response in older adults with or without subclinical depressive symptoms.MethodsThis cross-sectional study was conducted using baseline data from the Age-Well trial. Cognitively unimpaired (CU) older adults with (DepS; n = 73) or without (NoDepS; n = 58) subclinical depressive symptoms (defined using the 15-item Geriatric Depression Scale) were included in the analyses. Blood cortisol, epinephrine and norepinephrine were measured; as well as the resting-state functional connectivity (rs-FC) between, and gray matter (GM) volume of, the hypothalamus, hippocampus and hippocampal subfields. Blood stress markers levels and neuroimaging measures were compared between groups; then regression analyses were conducted between these measures.ResultsDepS participants showed higher plasma epinephrine levels, which was associated with greater rs-FC between the CA1 and Subiculum hippocampal subfields and the hypothalamus. Lower GM volume in the CA1 and DG/CA2-3–4 subfields was also found in DepS. No between-group differences were observed for blood cortisol and norepinephrine.ConclusionsOur findings show that subclinical depressive symptoms are associated with increased sympatho-adrenomedullary axis activity, together with lower GM volume in a hippocampal subfield (i.e., CA1) particularly sensitive to AD. While causation cannot be inferred, these results suggest that screening and treating subclinical depressive symptoms in CU older adults could reduce AD risk.Trial registrationClinicalTrials.gov Identifier: NCT02977819, Registration Date: 2016–11-25.

Network pharmacology-based strategy to reveal Acacetin against lipopolysaccharide-induced lung injury.

Acacetin, a flavonoid isolated from Agastache rugosa, exhibits diverse biological activities, such as anti-tumor, anti-inflammatory and antioxidant activities. Its role in treating Lipopolysaccharide (LPS)-induced acute lung injury (ALI) remains incompletely illuminated. To explore the potential molecular mechanisms of Acacetin in alleviating ALI. The network pharmacological approach was employed to screen the target genes and pathways of Acacetin. Lung injury was analyzed by Hematoxylin-Eosin (H&E) staining. Bronchoalveolar lavage fluid, serum and lung tissues were collected to detect the levels of proinflammatory cytokines and oxidative stress markers. Immunofluorescence and RT-qPCR experiments were used to observe the expression of CD45, COX2, Ly6G, and related-target proteins. In vitro, RAW264.7 macrophages were stimulated with LPS and treated with AMPK siRNA or an AMPK inhibitor Coumpound C to verify the role of AMPK/nuclear factor erythroid 2-related factor 2 (Nrf2)/high-mobility group box 1 (HMGB1) signaling in Acacetin-mediated alleviation of ALI. Network data revealed that Acacetin could regulate HMGB1, AMPK, Nrf2, and IL-6. In vivo, Acacetin reversed pathological damage and the release of inflammatory factors, and alleviated oxidative stress and immune cell infiltration in ALI development. Acacetin remarkably upregulated the expression of AMPK and Nrf2, accompanied by HMGB1 downregulation. In vitro, inhibiting AMPK reversed the effects of Acacetin in LPS-treated RAW264.7, due to inactivation of AMPK/Nrf2/HMGB1 pathway. The combination of network pharmacology and experimental studies revealed the role of Acacetin in improving ALI via the AMPK/Nrf2/HMGB1 signaling axis, which provided new insights into the treatment of ALI with Acacetin as a candidate drug.

Substituting animal protein with black soymilk reduces advanced glycation end product level and improves gut microbiota composition in obese prediabetic individuals: a randomized crossover intervention trial.

Prediabetes (PreDM) and obesity increase the risk of type 2 diabetes. Individuals with these conditions often consume diets higher in animal protein than in plant protein, which are associated with elevated levels of dietary advanced glycation end products (dAGEs). Increased dAGE intake has been linked to blood glucose abnormalities, oxidative stress, and dysbiosis of the microbiota, all of which exacerbate metabolic disorders. Black soybeans, as a plant-based protein source, contain substantially lower levels of dAGEs compared with pork. This study aimed to investigate the effects of substituting animal protein with black soybeans on advanced glycation end product (AGE) levels, oxidative stress, and the gut microbiota in individuals with both PreDM and obesity. This study was a randomized crossover intervention trial conducted over 16 weeks. We recruited men and women aged 20-64 years with both prediabetes and obesity. This study had four periods: 0-4 weeks for the run-in period, 4-8 weeks and 12-16 weeks for the pork or black soymilk intervention period, and 8-12 weeks for the wash-out period. During the intervention period, the participants consumed pork or black soymilk with similar protein content as their dietary protein source. The participants maintained 3 day dietary records, and we measured anthropometric items and collected blood and fecal samples for analysis. The results showed that partially substituting pork with black soymilk as a dietary protein source for 4 weeks significantly reduced dAGE intake. The black soymilk group also exhibited significantly lower blood AGE fluorescence intensity, oxidative stress, and levels of glycative stress markers. Furthermore, black soymilk consumption significantly increased the relative abundance of short-chain fatty acid-producing genera compared with pork consumption. In conclusion, partially substituting dietary pork with black soymilk may reduce serum AGE levels, reduce oxidative and glycation stress, and increase the abundance of short-chain fatty acid-producing microbiota in individuals with both PreDM and obesity. Registration number of Clinical Trial: NCT05290519 (ClinicalTrials.gov).

Pterostilbene attenuates oxidative stress induced by hydrogen peroxide in MAC-T cells through activating PINK1/Parkin-mediated mitophagy

High‐producing dairy cows are susceptible to altered redox balance owing to their high secretion and strong metabolism during transition to lactation. Previously reports demonstrated that pterostilbene (PTE) alleviate hydrogen peroxide (H2O2)-induced oxidative damage. However, molecular mechanism underlying protective role of PTE in H2O2-elicited oxidative stress in bovine mammary epithelial cells (MAC-T cells) remains unclear. The aims of our research is to clarify the potential molecular mechanisms of PTE in H2O2-triggered oxidative damage. MAC-T cells were pre-treated with PTE (0, 10, 25 and 50 μM) for 12 h and then cultured with H2O2 (0, 100, 200, 400, 600, 800 and 1000 μM) for another 24 h. The results were interpreted by CCK8, western blot, and immunofluorescence assay. Results showed that PTE significantly attenuated H2O2-mediated reduction in T-AOC, SOD and GSH-Px activity. Mechanistic studies found that PTE restricted the H2O2-induced protein expression of HO-1, GPX4 and SOD1, and PTE reversed intracellular MDA and ROS generation and decreased the MMP in MAC-T cells. Moreover, H2O2 exposure markedly inhibited mitophagy, whereas PTE pre-treatment activated PINK1/Parkin-mediated mitophagy pathway, which further limited ROS production. Overall, PTE can be used to improve oxidative stress and as a novel antioxidant agent in dairy cows. Highlights PTE restored the levels of H2O2-induced oxidative stress markers (T-AOC, SOD, GSH-Px and MDA) in MAC-T cells. PTE effectively blocked the increase of protein expression (HO-1, SOD1 and GPX4) in H2O2-induced MAC-T cells. PTE remarkably decreased the accumulation of ROS and improved MMP in H2O2-induced MAC-T cells. PTE exerted a significant protective effect against oxidative stress injury by reducing the inhibition of H2O2 on mitophagy.

CHANGES IN NEUROHUMORAL STATUS OF PATIENTS WITH COMMUNITY-ACQUIRED PNEUMONIA IN ASSOCIATION WITH CORONAVIRUS INFECTION COVID-19

The aim of this study was to analyze and summarize the available data on changes in neurohumoral status in patients with community-acquired pneumonia associated with COVID-19, as well as to identify the main mechanisms explaining these changes. Materials and methods. For a systematic review of the literature, scientific articles published in leading scientific databases, including PubMed, Scopus, Google Scholar and Web of Science, which provide access to peer-reviewed articles and other scientific publications, were selected. Results. Among patients with community-acquired pneumonia caused by COVID-19, the most common forms of involvement were bilateral multisegmental and right-sided lower lobe pneumonia. The localization of pneumonia varies significantly by gender and age group, with right-sided lower lobe pneumonia more common in men aged 20-30 and 51-60 years, and right-sided upper lobe pneumonia in women aged 20-30 years and left-sided lower lobe pneumonia in women aged 31-50 years. The most common forms in mild disease are left-sided and right-sided lower lobe pneumonia, while in severe disease, bilateral multisegmental pneumonia is the most common. Changes in the levels of immunoinflammatory markers (CRP, IL-6), the stress hormone cortisol, and the thrombosis marker D-dimer were important prognostic markers of disease severity. In patients with severe pneumonia, a significant increase in these parameters was observed, which is associated with a high risk of developing thromboembolic complications and required intensive care. Elevated levels of CRP, IL-6, D-dimer and cortisol in patients with severe COVID-19 are prognostic markers for assessing the severity of the disease and the development of complications such as thromboembolism. Conclusion. Thus, early detection of changes in the levels of inflammatory and stress markers allows predicting the severity of the disease, assessing the risk of complications, and timely applying treatment strategies to reduce mortality and improve treatment outcomes.

EZH2/miR-142-3p/HMGB1 axis mediates chondrocyte pyroptosis by regulating endoplasmic reticulum stress in knee osteoarthritis.

Knee osteoarthritis (OA) is still challenging to prevent or treat. Enhanced endoplasmic reticulum (ER) stress and increased pyroptosis in chondrocytes may be responsible for cartilage degeneration. This study aims to investigate the effect of ER stress on chondrocyte pyroptosis and the upstream regulatory mechanisms, which have rarely been reported. The expression of the histone methyltransferase enhancer of zeste homolog 2 (EZH2), microRNA-142-3p (miR-142-3p), and high mobility group box 1 (HMGB1) and the levels of ER stress, pyroptosis, and metabolic markers in normal and OA chondrocytes were investigated by western blotting, quantitative polymerase chain reaction, immunohistochemistry, fluorescence in situ hybridization, fluorescein amidite-tyrosine-valine-alanine-aspartic acid-fluoromethyl ketone (FAM-YVAD-FMK)/Hoechst 33342/propidium iodide (PI) staining, lactate dehydrogenase (LDH) release assays, and cell viability assessments. The effects of EZH2, miR-142-3p, and HMGB1 on ER stress and pyroptosis and the hierarchical regulatory relationship between them were analyzed by chromatin immunoprecipitation, luciferase reporters, gain/loss-of-function assays, and rescue assays in interleukin (IL)-1β-induced OA chondrocytes. The mechanistic contribution of EZH2, miR-142-3p, and HMGB1 to chondrocyte ER stress and pyroptosis and therapeutic prospects were validated radiologically, histologically, and immunohistochemically in surgically induced OA rats. Increased EZH2 and HMGB1, decreased miR-142-3p, enhanced ER stress, and activated pyroptosis in chondrocytes were associated with OA occurrence and progression. EZH2 and HMGB1 exacerbated and miR-142-3p alleviated ER stress and pyroptosis in OA chondrocytes. EZH2 transcriptionally silenced miR-142-3p via H3K27 trimethylation, and miR-142-3p posttranscriptionally silenced HMGB1 by targeting the 3'-UTR of the HMGB1 gene. Moreover, ER stress mediated the effects of EZH2, miR-142-3p, and HMGB1 on chondrocyte pyroptosis. In vivo experiments mechanistically validated the hierarchical regulatory relationship between EZH2, miR-142-3p, and HMGB1 and their effects on chondrocyte ER stress and pyroptosis. A novel EZH2/miR-142-3p/HMGB1 axis mediates chondrocyte pyroptosis and cartilage degeneration by regulating ER stress in OA, contributing novel mechanistic insights into OA pathogenesis and providing potential targets for future therapeutic research.

Limonin alleviates imiquimod-induced psoriasis-like skin inflammation in mice model by downregulating inflammatory responses.

Psoriasis is a chronic inflammatory condition affecting 1-2% of the global population. Phytomedicine, which uses plant-based compounds, is emerging as a promising approach to managing such inflammatory diseases. Limonin, a phytochemical found in citrus fruits and known for its bitter taste, possesses significant pharmacological properties. In this study, we evaluated the anti-psoriatic effects of limonin using a psoriasis-induced mice model. BALB/c mice were treated with imiquimod to induce psoriasis and then administered limonin at doses of 20 and 40 mg/kg/day for 6 days. Tacrolimus ointment served as a positive control. We assessed the hematological profile to determine limonin's impact on leukocytes in the psoriasis model. Additionally, histomorphometric analysis of ear and skin tissues was conducted to evaluate the therapeutic effects of limonin. We further investigated the antioxidant properties of limonin by measuring levels of antioxidants and oxidative stress markers. The anti-inflammatory effects were evaluated by quantifying inflammatory cytokines and signaling proteins. In vitro, the cytotoxicity and anti-inflammatory potential of limonin were assessed using murine macrophage RAW264.7 cells. Our findings showed that limonin significantly reduced leukocyte counts, decreased inflammatory cell infiltration, and improved skin histoarchitecture in psoriasis-induced mice. Limonin also effectively scavenged free radicals and reduced levels of inflammatory cytokines and proteins without causing cytotoxicity in RAW264.7 cells. Overall, our in vivo and in vitro results confirm that limonin is a potent anti-inflammatory agent that effectively ameliorates imiquimod-induced psoriasis.

MiR-378 exaggerates angiogenesis and bone erosion in collagen-induced arthritis mice by regulating endoplasmic reticulum stress

Rheumatoid arthritis (RA) is a chronic autoimmune disorder marked by pain, inflammation, and discomfort in the synovial joints. It is critical to understand the pathological mechanisms of RA progression. MicroRNA-378 (miR-378) is highly expressed in the synovium of RA patients and positively correlated with disease severity, but its function and underlying mechanisms remain poorly understood. In this study, miR-378 transgenic (miR-378high) mice were used to construct the collagen-induced arthritis (CIA) model for exploring the role of miR-378 in RA development. miR-378high CIA mice showed accelerated RA development, as evidenced by exaggerated joint swelling and bone structural deformities. More severe endoplasmic reticulum (ER) stress and the consequent angiogenesis and osteoclastogenesis were also activated in the synovial tissue and calcaneus, respectively, in the miR-378high group, suggesting that ER plays a significant role in miR-378-mediated RA pathogenesis. Upon in vitro RA induction, fibroblast-like synoviocytes (FLSs) isolated from miR-378high mice showed a higher expression level of ER stress markers. The conditioned medium (CM) from RA-FLSs of miR-378high mice stimulated more intensive angiogenesis and osteoclastogenesis. The ER stress-related protein Crebrf was identified as a downstream target of miR-378. Crebrf knockdown diminished the promoting effect of miR-378 on ER stress, as well as its downstream angiogenesis and osteoclastogenesis activities. Tail vein injection of anti-miR-378 lentivirus in an established RA mouse model was shown to ameliorate RA progression. In conclusion, miR-378 amplified RA development by promoting ER stress and downstream angiogenesis and osteoclastogenesis, thus indicating that miR-378 may be a potential therapeutic target for RA treatment.

Individual muscle hypertrophy in high-load resistance training with and without blood flow restriction: A near-infrared spectroscopy approach

ABSTRACT We aimed to compare individual hypertrophic responses to high-load resistance training (HL-RT) or high-load with blood flow restriction (HL-BFR). Furthermore, we investigated whether greater responsiveness to one of the protocols could be explained by acute changes in blood deoxyhemoglobin concentration (HHb) and total hemoglobin concentration (tHb) (proxy markers of metabolic stress). Ten untrained participants had their legs randomized into both HL-RT and HL-BFR and underwent 10 weeks of training. Muscle cross-sectional area (mCSA) was measured at baseline and post training, while HHb and tHb during the final session. Using a threshold of 2 × typical errors (3.24%) to compare protocols, five participants showed greater mCSA increases after HL-RT (16.44 ± 7.90%) compared to HL-BFR (10.74 ± 7.12%, p = 0.0054) and five did not respond better to HL-RT (8.95 ± 10.83%) compared to HL-BFR (13.33 ± 8.59%) (p = 0.3105). Additionally, HL-RT induced lower HHb (5855.78 ± 12905.99; p = 0.0101) and tHb (−43169.70 ± 37793.17; p = 0.0030) AUC values compared to HL-BFR (HHb: 39254.80 ± 27020.15; tHb: 46309.40 ± 31613.97). In conclusion, despite the higher levels of metabolic stress markers, most participants did not present greater muscle hypertrophy by combining blood flow restriction with HL-RT.

Entacapone alleviates muscle atrophy by modulating oxidative stress, proteolysis, and lipid aggregation in multiple mice models

BackgroundSkeletal muscle atrophy significantly affects quality of life and has socio-economic and health implications. This study evaluates the effects of entacapone (ENT) on skeletal muscle atrophy linked with oxidative stress and proteolysis.MethodsC2C12 cells were treated with dexamethasone (Dex) to simulate muscle atrophy. Four murine models were employed: diaphragm atrophy from mechanical ventilation, Dex-induced atrophy, lipopolysaccharide (LPS)-induced atrophy, and hyperlipidemia-induced atrophy. Each model utilized entacapone (10 mg/kg), with sample sizes: Control (9), MV (11), MV + ENT (5) for diaphragm atrophy; Control (4), Dex (4), Dex + ENT (5) for Dex model; Control (4), LPS (4), LPS + ENT (5) for LPS model; and similar for hyperlipidemia. Measurements included muscle strength, myofiber cross-sectional area (CSA), proteolysis, oxidative stress markers [uperoxide dismutase 1 (SOD1), uperoxide dismutase 2 (SOD2), 4-hydroxynonenal (4-HNE)], and lipid levels.ResultsOur findings confirm Dex-induced muscle atrophy, evidenced by increased expression of muscle atrophy-associated proteins, including Atrogin-1 and Murf-1, along with decreased diameter of C2C12 myotubes. Atrogin-1 levels rose by 660.6% (p &amp;lt; 0.05) in the Dex group compared to control, while entacapone reduced Atrogin-1 by 84.4% (p &amp;lt; 0.05). Similarly, Murf-1 levels increased by 365% (p &amp;lt; 0.05) in the Dex group and were decreased by 89.5% (p &amp;lt; 0.05) with entacapone. Dexamethasone exposure induces oxidative stress, evidenced by the upregulation of oxidative stress-related proteins Sod1, Sod2, and 4-HNE. Entacapone significantly reduced the levels of these oxidative stress markers, enhancing GSH-PX content by 385.6% (p &amp;lt; 0.05) compared to the Dex-treated group. Additionally, ENT effectively reduced the Dex-induced increase in MDA content by 63.98% (p &amp;lt; 0.05). Furthermore, entacapone effectively prevents the decline in diaphragm muscle strength and myofiber CSA in mice. It also mitigates diaphragm oxidative stress and protein hydrolysis. Additionally, entacapone exhibits the ability to attenuate lipid accumulation in the gastrocnemius muscle of hyperlipidemic mice and alleviate the reduction in muscle fiber CSA.ConclusionOur findings suggest that entacapone is a promising therapeutic candidate for muscle atrophy, functioning through the reduction of oxidative stress, proteolysis, and lipid aggregation. Future research should explore the underlying mechanisms and potential clinical applications of entacapone in muscle-wasting conditions.

Protective effects of naringenin against methamphetamine-induced cell death in dopaminergic SH-SY5Y cells

ABSTRACT Background: Methamphetamine is a psychoactive substance that competes with the dopamine transporter, disrupting its flow and storage. This can trigger oxidative stress, finally resulting in neural cell death. Due to the increasing prevalence of methamphetamine use, extensive research has been devoted to finding treatments that ameliorate its detrimental effects. Naringenin, a dietary flavonoid found in citrus fruits, has shown several neuroprotective and pharmacological properties. Objectives: This study was aimed to assess the protective effects of naringenin against methamphetamine-induced dopaminergic cell death. Methods: Before exposure to methamphetamine, human neuroblastomaSH-SY5Y cells were either pretreated or not treated (controls) with naringenin. Cell viability, level of oxidative stress markers, and expression of some genes involved in apoptosis and autophagy processes were then assessed using MTT, ROS, and MMP assays, and qRT-PCR and Western blotting techniques. Results: Naringenin pretreatment significantly enhanced cell viability following methamphetamine exposure (p < .01). It significantly decreased ROS levels (p < .001), preserved mitochondrial membrane potential, and moderated upregulation of apoptotic (CytC, Casp3, and Bax) and autophagic genes (Beclin-1, and LC-3) and down-regulation of Bcl-2 as an anti-apoptotic gene. Similar naringenin-mediated patterns were observed for cytochrome C and caspase 3 proteins. Conclusion: Naringenin administration can be considered for treating the neurotoxic effects of methamphetamine.

Effect of aqueous extract of Mondia whitei fruits on high sucrose-induced oxidative stress in Drosophila melanogaster

Background/Aim: High sugar intake plays an important role in oxidative stress, which has been linked to development of diseases like diabetes. In this study, the effect of aqueous extract of Mondia whitei (AEMW) fruits on high sucrose-induced oxidative stress in Drosophila melanogaster was investigated. Materials and Methods: Adult flies (1 – 3 days old) were exposed to graded doses of AEMW for 12 days to determine the safe dose. Based on this, new flies were grouped into 5 (n = 5; 40 flies per vial). Group 1 served as control and were fed with basal diet, and group 2 with basal diet containing 30% sucrose. Groups 3 and 4, in addition to diet containing 30 % sucrose, were exposed to 0.25 and 0.5 mg AEMW/g diet, respectively, and group 5 flies were exposed to diet containing 0.5 mg AEMW/g diet only. Treatment lasted for 7 days. Biochemical assays such as glucose level, markers of antioxidant and oxidative stress were carried out to assess the effect of high sucrose and/or AEMW on flies. Results: Doses of AEMW up to 0.5 mg/g diet was safe for the flies. High sucrose diet significantly (p &lt; 0.05) increased the level of glucose and oxidative stress markers, and reduced the levels of antioxidant parameters when compared to the control. These changes were prevented by the doses of AEMW. Conclusion: It can be concluded that AEMW possesses anti-hyperglycemic and antioxidant properties, and could be suggested to act against oxidative stress induced by high sucrose in fruit flies.

Protective effects of insoluble dietary fiber from cereal bran against DSS-induced chronic colitis in mice: From inflammatory responses, oxidative stress, intestinal barrier, and gut microbiota.

Insoluble dietary fiber (IDF) is a crucial component of cereals, and IDF from cereal bran (IDF-CB) has been reported to have multiple biological activities. However, the effect of IDF-CB on chronic colitis remains underexplored. The study aimed to investigate the impact of IDFs from wheat bran (WBIDF), rice bran (RBIDF), millet bran (MBIDF) and oat bran (OBIDF) on chronic colitis induced by dextran sulfate sodium (DSS). Our findings demonstrated that IDFs-CB supplementation mitigated DSS-induced weight loss and reduced lesions in the colon and spleen. Levels of proinflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) and oxidative stress markers (MPO, iNOS and MDA)were decreased, and anti-inflammatory cytokine (IL-10) and T-SOD activity were increased after IDF-CB inclusion. Furthermore, IDFs-CB restored intestinal barrier function by regulating gene expression (up-regulated Muc-2, ZO-1 and Occludin, and down-regulated Claudin-1 and Claudin-4). Additionally, we analyzed the gut microbiota and SCFAs composition. WBIDF, MBIDF and OBIDF inhibited the growth of Muribaculaceae_unclassified, Bacteroides and Parasutterella. Conversely, IDFs-CB promoted the growth of Candidatus_Saccharimonas and norank_f__norank_o__Clostridia_UCG-014. Notably, WBIDF enhanced the abundance of Allobaculum, while MBIDF and OBIDF increased the abundance of Lachnospiraceae_NK4A136. Moreover, supplementation with IDFs-CB significantly elevated certain SCFA concentrations-particularly acetic acid and isohexanoic acid. Our results suggested that IDF-CB effectively alleviated DSS-induced chronic colitis; among them，WBIDF exhibited superior efficacy followed by OBIDF，MBIDF，and RBIDF. This study provides a theoretical foundation for dietary recommendations for patients suffering from inflammatory bowel disease.

Ameliorating Effect of S-Allyl Cysteine (Black Garlic) on 6-OHDA Mediated Neurotoxicity in SH-SY5Y Cell Line

Therapeutic approaches based on isolated compounds derived from natural products are more common in preventing diseases involving inflammation and oxidative stress at present. S-allyl cysteine (SAC) is a promising garlic-derived organosulfur compound with many positive effects in cell models and living systems. SAC has biological activity in various fields, enclosing healing in learning and memory disorders, neurotrophic effects, and antioxidant activity. In this study, we purposed to identify the neuroprotective activity of SAC toward 6-OHDA-induced cell demise in the SH-SY5Ycell line. For this purpose, 6-OHDA-induced cytotoxicity, and biochemical, and gene expression changes were evaluated in SH-SY5Y cells. SH-SY5Y cells grown in cell culture were treated with SAC 24hours before and after 6-OHDA application. Then, cell viability, antioxidant parameters, and gene expressions were measured. Finally, immunofluorescence staining analysis was performed. Our results showed that SAC increased cell viability by 144% at 80µg/mL with pre-incubation (2hours). It was observed that antioxidant levels were significantly increased and oxidative stress marker levels were decreased in cells exposed to 6-OHDA after pre-treatment with SAC (p<0.05). SAC supplementation also suppressed the increase in pro-inflammation levels (TNF-α/IL1/IL8) caused by 6-OHDA (p < 0.05). While 8-OHdG and Nop10 expressions were observed at a mild level in SAC pretreatment depending on the dose, 8-OHdG, and Nop10 expressions were observed at a moderate level in SAC treatment after 6-OHDA application (p<0.05). Our findings demonstrate the positive effect of pretreatment with SAC on SH-SY5Y cells injured by 6-OHDA, suggesting that SAC may be beneficial for neuroprotection in regulating oxidative stress and neuronal survival in an in vitro model of Parkinson's disease.

The therapeutic effects of curcumin on polycystic ovary syndrome by upregulating PPAR-γ expression and reducing oxidative stress in a rat model.

Polycystic ovary syndrome (PCOS) is a prevalent endocrine and metabolic disorder that impacts 8-13% of women in their reproductive years. However, the drugs commonly used to treat PCOS are often prescribed off-label and may carry potential side effects. This study aimed to investigate the therapeutic effects of curcumin in a PCOS rat model. A PCOS rat model was established through daily subcutaneous injection of 60 mg/kg body weight of dehydroepiandrosterone (DHEA) for 21 days. The PCOS rats received a daily intragastric dose of 50 mg/kg body weight of curcumin for another 21 days. Ovarian morphological changes, estrous cycle changes, and hormone level changes were measured to evaluate the therapeutic effectiveness of curcumin in PCOS rats. Oxidative stress markers in the ovaries were analyzed to explore the mechanisms of curcumin in PCOS rats. This study demonstrated that curcumin alleviated insulin resistance and significantly reduced serum levels of estradiol (p = 0.02), luteinizing hormone (p = 0.009), testosterone (p = 0.003), and the LH/FSH ratio (p = 0.008) in PCOS rats. Curcumin also restored normal ovarian morphology and the estrous cycle in these rats. Furthermore, curcumin treatment significantly decreased levels of oxidative stress markers, including malondialdehyde (p = 0.004) and reactive oxygen species (p = 0.005), while increasing antioxidant levels such as superoxide dismutase (p = 0.04), glutathione peroxidase (p = 0.002), and glutathione (p = 0.02) in ovarian tissues. Additionally, curcumin significantly upregulated PPAR-γ in the ovarian tissues of PCOS rats. This study demonstrates that curcumin effectively restores ovarian morphology, hormone levels, and estrous cycles in PCOS rats. These effects may be linked to its ability to reduce oxidative stress in ovaries via the upregulation of PPAR-γ. Curcumin shows promise as a potential drug for the treatment of PCOS.