Oxidative Markers Research Articles

Antioxidant supplementation boosts the advantages of CrossFit workouts on oxidative and muscle damage markers in obese males.

Supplementing with antioxidants may be one of the most efficient means of minimizing oxidative stress during workouts in obese individuals. The aim of this study is to identify the results after twelve weeks of CrossFit workouts combined with Spinach thylakoid extract on the levels of insulin resistance (insulin and Homeostatic Model Assessment of Insulin Resistance [HOMA-IR]), fasting blood sugar (FBS), malondialdehyde (MDA), creatine kinase (CK), lactate dehydrogenase (LDH), total antioxidant capacity (TAC), and superoxide dismutase (SOD), glutathione peroxidase (GPx) in obese males. Sixty-eight males with an average age of 27 ± 8 yrs and a BMI of 32.6 ± 2.6kg.m- 2 were randomly split into four groups each consisting of seventeen individuals. : control group (CG), supplement group (SG), training group (TG), and training + supplement group (TSG). After initial assessments, the two training groups (TG and TSG) started on a 12 weeks of the CrossFit workouts program involving three sessions per week each lasting up to 60min. Participants in supplement groups ingested 30min before lunch, 5 gof Spinach thylakoid extract per day or one sachet of raw corn starch in the control group. Baseline and post-intervention measurements were performed 48h pre- and post-last session, respectively. The findings revealed noteworthy relationships between the exercise groups and timefor TAC, SOD, GPx, MDA, CK, and LDH (p < 0.001, ES: 0.88, 0.88, 0.8, 0.4, 0.7, and 0.7, respectively). In addition, there were statistically significant differences among study groups after attending the intervention program in TAC (ES: 0.88), SOD (ES: 0.92), GPX (ES: 0.85), MDA (ES: 0.5), CK (ES: 0.7) and LDH (ES: 0.8). The effect sizes of insulin (0.77), glucose (0.21), and HOMA-IR (0.44) varied significantly (p < 0.05) among the groups. The results demonstrated that CrossFit workouts for 12 weeks combined with Spinach thylakoid extract in men with obesity may prevent oxidative damage caused by obesity and CrossFit workouts.

Embryonic Lethal Abnormal Visual-Like Protein 1 Aggravates Caerulein-Induced AR42J Cell Injury and Macrophage M1 Polarization to Accelerate Acute Pancreatitis by Upregulating TRAF6.

Tumor necrosis factor receptor-associated factor 6 (TRAF6) has been found to promote the progression of acute pancreatitis (AP). However, its underlying molecular mechanisms in AP need to be further revealed. Caerulein-induced AR42J cells were used to construct AP cell models. Cell viability and apoptosis were measured by Cell Counting Kit 8 assay and flow cytometry. Levels of inflammatory factors and oxidative stress-related markers were assessed. The medium of AR42J cells was collected for coculturing RAW264.7 cells. Macrophage marker CD86+ cell rates were checked with flow cytometry. The levels of TRAF6, embryonic lethal abnormal visual-like protein 1 (ELAVL1), and inducible nitric oxide synthase (iNOS) were examined by Western blot or quantitative real-time polymerase chain reaction. RNA immunoprecipitation assay was performed to evaluate the interaction between ELAVL1 and TRAF6. TRAF6 mRNA stability was tested using actinomycin D treatment. Caerulein treatment suppressed viability, induced AR42J cell apoptosis, inflammation, oxidative stress, and accelerated macrophage M1 polarization. TRAF6 downregulation could alleviate caerulein-induced AR42J cell injury and macrophage M1 polarization. ELAVL1 interacted with TRAF6 to stabilize its expression. Meanwhile, ELAVL1 knockdown relieved caerulein-induced AR42J cell injury and macrophage M1 polarization, while these effects were abolished by TRAF6 overexpression. TRAF6, stabilized by ELAVL1, promoted caerulein-induced AR42J cell injury and macrophage M1 polarization, suggesting that it might accelerate AP9 progression.

Abstract 4134707: Metabolomics Discovery and Epicardial Adipose Tissue : a PROMISE Clinical Trial Substudy

Introduction: Epicardial adipose tissue (EAT) is a metabolically active tissue associated with cardiometabolic disease. EAT likely has local paracrine inflammatory and atherogenic effects on coronary vessels, but the metabolic underpinnings related to systemic metabolic pathways has not been evaluated. Hypothesis: We hypothesized that metabolic markers of lipid and fatty acid oxidation will be positively associated with volume of EAT. Methods: In a biomarker substudy of the Prospective Multicenter Imaging Study for Evaluation of Chest Pain (PROMISE) clinical trial which randomized participants to computed tomography angiography (CTA) vs. stress testing, we analyzed 1542 participants randomized to the CTA arm with available plasma at baseline. CTA phenotyping of EAT volume indexed to body surface area (cm 3 /m 2 ) was determined using a validated deep-learning algorithm. Comprehensive metabolomic profiling was performed on plasma samples using mass spectrometry. Principal components analysis (PCA) was used for dimensionality reduction of 571 named metabolites. Linear regression tested the association of metabolomic factors with EAT. Univariate models were adjusted for multiple comparisons and multivariate models were adjusted for age, race and ethnicity, sex, diabetes, BMI, LDL-C, HDL-C and metabolic syndrome. Results: Of 82 PCA-derived metabolomic factors, 16 were associated with EAT in univariate models and nine were associated with EAT in multivariate models (p=0.001-0.046); 116 individual metabolites heavily loaded within these factors were significantly associated with EAT volume in multivariate models ( Figure 1 ). This included three linoleic acid derivatives, six cholesterol esters, and glycine that were inversely associated with EAT volume (p&lt;0.04) and 106 metabolites positively associated with EAT volume including di- and triacylcerols, glycerophospholipids, fatty acyls and dimethylguanidino valeric acid (p&lt;0.05). Conclusions: Leveraging a large clinical trial of CTA to study metabolomics of EAT, we have identified circulating lipid species associated with EAT independent of BMI and conventional lipids. These findings highlight biomarkers of systemic metabolic dysregulation as risk modifiers and therapeutic targets in cardiometabolic disease prevention and treatment.

Ribes khorasanicum: A Potent Antioxidant Against Organ Toxicity by Effect on the NF‐κB Pathway

ABSTRACTAcetaminophen (APAP) is a well‐known drug that, in high doses, induces hepatotoxicity and nephrotoxicity. This study has investigated the preventive effect of the extract and fractions of Ribes khorasanicum on APAP‐induced liver and kidney damage. In this experiment, after analysis of the extract using FTIR, toxicity was induced by APAP on the 7th day. Before that, the extract and their aqueous, ethyl acetate, and n‐hexane fractions were administrated orally. 24 h after APAP administration, the animals were sacrificed. The liver and kidney were removed for the evaluation of oxidation and biochemical markers, including SGOT, SGPT, ALP, LDH, albumin, urea, creatinine, and bilirubin and also for histopathological evaluations. The safety of the extract was evaluated based on the MTT. Also, expression of the NF‐kB is done. Administration of Ribes khorasanicum significantly decreased the biochemical parameters compared to the APAP. Oxidative parameters, as well as histopathological changes in preventive groups, were improved compared to the APAP in both tissues. The results showed that the aqueous and ethyl acetate fractions of the extract had a better effect than the whole extract and n‐hexane fraction against APAP toxicity.

Evaluation of the cell death markers for aberrated cell free DNA release in high altitude pulmonary edema.

The effect of high altitude (HA, altitude >2500 m) can trigger a maladaptive response in unacclimatized individuals, leading to various HA illnesses such as high altitude pulmonary edema (HAPE). The present study investigates circulating cell free (cf) DNA, a minimally invasive biomarker that can elicit a pro-inflammatory response. Our earlier study observed altered cfDNA fragment patterns in HAPE patients and the significant correlation of these patterns with peripheral oxygen saturation levels. However, the unclear release mechanisms of cfDNA in circulation limit its characterization and clinical utility. The present study not only observed a significant increase in cfDNA levels in HAPE patients (27.03 ± 1.37 ng/ml; n = 145) compared to healthy HA sojourners (controls, 14.57 ± 0.74 ng/ml; n = 65) and highlanders (HLs, 15.50 ± 0.8 ng/ml; n = 34) but also assayed the known cell death markers involved in cfDNA release at HA. The study found significantly elevated levels of the apoptotic marker, annexin A5, and secondary necrosis or late apoptotic marker, High mobility group box 1, in HAPE patients. In addition, we observed ahigher oxidative DNA damage marker, 8-hydroxy-2'-deoxyguanosine, in HAPE compared to controls, suggestive of the role of oxidative DNA status in promoting the inflammatory potential of cfDNA fragments and their plausible role in manifesting HAPE pathophysiology. Extensive in vitro future assays can confirm theimmunogenic role of cfDNA fragments that may act as a danger-associated molecular pattern and associate with markers of cellular stresses in HAPE.

Thermal-Induced Alterations in Phenolic and Volatile Profiles of Monovarietal Extra Virgin Olive Oils.

In the present study, the influence of heating on the evolution of oxidative indices, antioxidant activity, phenolic and volatile compounds in monovarietal extra virgin olive oils (EVOOs) obtained from Leccino, Istarska bjelica, and Buža cultivars was investigated. The samples were submitted to heating in an air oven (180 °C and 220 °C), simulating usual roasting conditions typical for Mediterranean cuisine. The decreases in the oxidative indicators, phenolic and volatile compounds were more pronounced at higher heating temperatures, underlining the temperature dependency of the oxidative degradation during heating conditions. Despite this, it must be emphasized that a significant amount of phenolic compounds and antioxidative activity remained preserved after the heating treatment. Each oil cultivar showed some specificity during the course of the thermal degradation. Hydroxytyrosol acetate among phenolic compounds and octanal, (E)-2-octenal, hexanal, 3-pentanone, and 1-penten-3-one among the volatiles were underlined as possible markers of thermal oxidation. Principal component analysis revealed that the content of volatile compounds in monovarietal EVOO samples distinguished samples primarily by the heating temperature, while the changes in the phenolic compounds were cultivar-dependent aside from being influenced by the temperature of heating.

Serum oxidative markers and delta neutrophil index in hyperemesis gravidarum

Objectives: To evaluate the relationship between different serum oxidative markers and the delta neutrophil index and hyperemesis gravidarum. Methods: One hundred pregnant women were enrolled in the study and divided into two groups. Group 1 included 50 women with hyperemesis gravidarum, while Group 2 (control group) included 50 pregnant women similar in age, gestational week, and body mass index. Serum oxidative markers and complete blood count inflammatory markers were compared. Results: Native thiol and total thiol were significantly lower in the Group 1 when compared with the control group (P=0.029 for native thiol; P=0.035 for total thiol). Moreover, ischemia-modified albumin (IMA) and catalase values were significantly higher in the Group 1 than in the control group (P=0.023 for IMA; P=0.021 for catalase). Index1% shows the disulfide/native thiol percent ratio and means that the Group 1 oxidant load is increased but not statistically significant. Myeloperoxidase, ferroxidase, and the delta neutrophil index did not differ significantly between the two groups (P=0.591, P=0.793, and P=0.52; respectively). Conclusions: According to our study, contrary to the literature, although there are differences in some values, when evaluated individually hyperemesis gravidarum does not impose an extra burden on maternal oxidant-antioxidant balance.

Effects of a Cyperus esculentus L. plant-based beverage on FVB/n female mice

This work evaluated the effects of the consumption of a plant-based beverage (Cyperus esculentus L., Adansonia digitata L., and thermal water) on the physiological parameters of mice over a 28-day period. Thirty-two female FVB/n mice (n = 8) were randomly assigned to one of four groups divided into two experimental protocols: Group 1 drank water and Group 2 drank a plant-based beverage for 24 hours; in the second experimental protocol, Group 3 consumed water and Group 4 consumed plant-based beverage for a limited time (4 hours). Two experimental protocols were conducted to assess whether the exposure time to the beverage affects the animals' physiological parameters, to determine if there is a possible daily limit for consumption of the product and to analyze their adverse effects. The mice consuming a beverage ingested a larger amount of drink and a smaller amount of food. Histologically there are no pathological changes in collected organs. The consumption of a plant-based beverage has been shown to have a positive effect on oxidative markers and can have a diuretic action. According to results, no behavioral changes or clinical signs of disease were observed throughout both experimental protocols, and no mortality was recorded.

20(R)-ginsenoside Rg3 Alleviates Diabetic Retinal Injury in T2DM Mice by Attenuating ROS-mediated ER Stress Through the Activation of the Nrf2/HO-1 Axis

BackgroundAlthough our previous work confirmed 20(R)-ginsenoside Rg3 (R-Rg3), which is an active ingredient in the Panax Ginseng C.A. Meyer, to have good anti-diabetic activity, its beneficial effect on diabetic retinal injury was found to be limited. PurposeThis study aims to investigate the protective effects of R-Rg3 on diabetes-induced retinal injury and the associated molecular mechanisms of action. MethodsDiabetic retinal injury was induced in mice using a combination of a high-fat diet (HFD) and intraperitoneal injection of streptozotocin (STZ). R-Rg3 (10 and 20 mg/kg) was subsequently administered for 6 weeks. The human retinal endothelial cells (HRECs) were subjected to high glucose (HG)-induced injury for the in vitro analysis and treated with R-Rg3 (4, 8, 16 μM), antioxidant N-Acetylcysteine (NAC, 1 mM) and Nrf2 inhibitor ML385 (5 μM). The mice retinas then underwent functional and histopathological analysis. Expression levels of proteins related to the Nrf2/HO-1 axis, tight junction proteins, endoplasmic reticulum (ER) stress and the apoptosis in retinal tissue and HRECs were determined by western blot. Expressions of ZO-1 and Nrf2 in the retina and HRECs were assessed by immunofluorescence. Additional evaluations included measuring body weights, fasting blood glucose (FBG), lipid levels and oxidative markers. ResultsThe results showed 6 weeks of R-Rg3 treatment significantly restored the functional changes and redox system imbalance that was induced by HFD/STZ in mice. R-Rg3 was also found to significantly reduce retinal barrier damage and thickness changes resulting from hyperglycaemia exposure. At the same time, R-Rg3 also protected HRECs from HG-induced damage. R-Rg3 could also activate Nrf2/HO-1 axis and inhibit endoplasmic reticulum stress as a means of alleviating retinal endothelial cells apoptosis. The molecular docking results also demonstrated that R-Rg3 had a good binding ability with Nrf2. ConclusionOur study suggested Nrf2/HO-1 axis might be crucial for the ability of R-Rg3 to prevent diabetic retinal injury.

Antipsychotic-Treated Schizophrenia Patients Develop Inflammatory and Oxidative Responses Independently From Obesity: However, Metabolic Disturbances Arise From Schizophrenia-Related Obesity.

To define the impact of obesity on inflammatory and oxidative disturbances in antipsychotic-treated schizophrenia patients. Several cytokines, inflammatory, metabolic, and oxidative status markers were evaluated in obese (n=40) and non-obese (n=40) antipsychotic-treated patients and compared with age-and BMI-matched controls (n=80). Schizophreniapatients had higher leptin, TNF-α, adiponectin, visfatin, resistin, P-selectin, NPY, BDNF, CD40-L, MCP-1, and malondialdehyde, and lower IL-6, ghrelin, neopterin, and vitamin E levels compared to their respective controls (p<0.001). Total oxidant status was higher in non-obese patients compared to controls (p<0.001), total antioxidant capacity was higher in obese compared to non-obese patients (p<0.01), but vitamin A and paraoxonase levels were not different. High sensitive-CRP levels were higher in obsese controls relative to non-obese controls (p<0.05) and in obese patients relative to non-obese patients (p<0.001). Fasting glucose, insulin, HbA1c, HOMA-IR, uric acid, total cholesterol, and triglyceride concentrations were higher in obese patients compared to non-obese patients. Insulin concentrations and HOMA-IR were also higher in obese controls than in non-obese controls. Our results suggest that inflammatory responses and oxidative stress develop independently from obesity in antipsychotic-treated schizophrenia patients. However, schizophrenia-induced obesity causes metabolic disturbances; thereby, obese schizophrenia patients are more liable to cardiovascular events and progress of metabolic syndrome than non-obese patients.

Protective effect of betaine supplementation in low-met diet against Aeromonas hydrophila-induced intestinal injury in grass carp (Ctenopharyngodon idella)

This study evaluated the effect of supplemental betaine in low-methionine (Met) diet concerning the regulatory role of bacteria-induced intestinal injury in fish. 540 grass carp (Ctenopharyngodon Idella) (210 ± 0.68 g) were selected and randomly divided into 6 groups, which contained a reference group fed with the diet supplemented with DL-Met (3.1 g kg−1 diet) and 5 groups fed on the diets added with increasing levels of betaine (0–6.4 g kg−1 diet) for 60 days. Then, a 14-day Aeromonas hydrophila challenge was conducted. Compared with BET0 group, results indicated that appropriate betaine supplementation (1) enhanced antioxidant enzymes activities (except glutathione S-transferase pi-1 (GSTP1) and copper zinc superoxide dismutase (CuZnSOD) and glutathione (GSH) contents, and NF-E2-related-factor 2 (Nrf2) protein nuclear translocation, while reduced reactive oxygen species (ROS), and representative oxidative damage markers; (2) up-regulated the transcription abundance of anti-apoptosis-related molecules while reduced DNA-ladder occurrence and the transcription levels of pro-apoptosis-related genes; (3) down-regulated the transcription of tight junction-related proteins (such as zonula occludens-1 (ZO-1) etc.) in fish intestines. Furthermore, correlation analysis revealed that feeding betaine ameliorating the oxidative damage, apoptosis and tight junction breakdown was possibly related to intestinal Nrf2, p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal kinases (JNK), and myosin light chain kinase (MLCK) signaling, respectively. According to the regression analysis for intestinal ROS abundance, claudin-b and caspase-2 transcription, the appropriate betaine supplementations were estimated to be 3.82, 3.98, and 3.62 g kg−1 diet, respectively. Notably, betaine has the potential to spare part Met and the efficacy of betaine relative to Met was 206.67 % based on intestinal ROS contents in grass carp (210–776 g).

Antineoplastic with DNA fragmentation assay and anti-oxidant, anti-inflammatory with gene expression activity of Lactobacillus plantarum isolated from local Egyptian milk products

Many lactic acid bacteria (LAB), known for their human health benefits, are derived from milk and utilized in biotherapeutic applications or for producing valuable nutraceuticals. However, the specific role of milk-associated LAB in biotherapeutics remains underexplored. To address this, eight milk product samples were randomly selected from the Egyptian market, diluted, and then cultured anaerobically on MRS agar. Subsequently, 16 suspected LAB isolates were recovered and underwent rapid preliminary identification. Among these isolates, the Lactobacillus plantarum strain with accession number (OQ547261.1) was identified due to its strong antioxidant activity depending on the DPPH assay, L. plantarum displayed notable antioxidant activities of 71.8% and 93.8% at concentrations of 125–1000 µg/mL, respectively. While ascorbic acid showed lower concentrations of 7.81, 3.9, and 1.95 µg/mL which showed activities of 45.1%, 34.2%, and 27.2%, respectively. The anti-inflammatory efficacy of L. plantarum was evaluated based on its capability to prevent hemolysis induced by hypotonic conditions. At a concentration of 1000 µg/mL, L. plantarum could reduce hemolysis by 97.7%, nearly matching the 99.5% inhibition rate achieved by the standard drug, indomethacin, at an identical concentration. Moreover, L. plantarum exhibited high hemolytic activity at 100 µg/mL (14.3%), which decreased to 1.4% at 1000 µg/mL. The abundance of phenolic acids and flavonoids was determined by high-performance liquid chromatography (HPLC) in L. plantarum. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) demonstrated that L. plantarum increased gene expression of the inflammatory marker TLR2 by 133%, and cellular oxidation markers SOD1 and SOD2 by 65% and 74.2%, respectively, while suppressing CRP expression by 33.3%. These results underscore L. plantarum’s exceptional anti-inflammatory and antioxidant activities. Furthermore, L. plantarum induces cancer cell death through necrotic nuclear DNA fragmentation. These findings suggest that L. plantarum is not only suitable for nutraceutical production but also holds potential as a probiotic strain. Future research should focus on enhancing the capacity of this strain across various industries and fostering innovation in multiple fields.

Radioprotective effects of melatonin therapy against testicular oxidative stress: a systematic review and meta-analysis of rodent models

Background: Radiation exposure is a concern in today’s world, given the widespread use of electronic devices and medical procedures involving ionizing and non-ionizing radiation. Radiations may cause male infertility by inducing oxidative stress in testicular tissue. Melatonin has antioxidant properties. Methods: We systematically reviewed the literature for the studies that have investigated the effects of melatonin therapy on radiation-induced oxidative stress in rodents’ testicular tissue. PubMed, Scopus, and Web of Science were searched for relevant animal trials. Standardized mean difference and 95% confidence intervals were used to pool the data. Subgroup and sensitivity analyses were done. The risk of bias was assessed using SYRCLE tool. Results: Outcomes: histopathology and sperm analyses (testicular apoptotic cells, Johnsen’s testicular biopsy score, seminiferous epithelial height, tubular diameter, sperm motility, viability, count, and morphology, concentration of spermatid, spermatocyte, and spermatogonia), body and testes weights (absolute and relative body and testicular weights), reproductive hormones (serum prolactin, FSH, and testosterone), and oxidative stress tissue markers (TBARS, CAT, GSH, GSH-Px, MDA, SOD, and XO, and total antioxidant capacity). Rats and mice were exposed to electromagnetic radiations (gamma, roentgen, microwave, radiofrequency, and high-power line energy) and particle waves (radioiodine and carbon-ion). Melatonin therapy was significantly associated with improved male reproduction. Conclusion: Radiation exposure harms male fertility, but melatonin, as an antioxidant, is potentially associated with improved male reproductive function in rodents. Inconsistencies in research require further investigations.

The effect of major depressive disorder comorbidity on ischemia-modified albumin levels, a marker of oxidative stress, and antioxidant defense system in patients with obsessive compulsive disorder.

The aim of this study was to examine the levels of oxidant and antioxidant markers in patients with obsessive-compulsive disorder (OCD) and to investigate whether these levels change in the presence of major depressive disorder (MDD) comorbidity. This study was completed with 23 OCD patients with MDD comorbidity (OCD+MDD), 21 OCD patients without MDD comorbidity (OCD-MDD) and 21 healthy controls. Oxidative stress levels of the cases' were determined by ischemia modified albumin (IMA) and malondialdehyde (MDA) measurements and antioxidant levels were determined by superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) measurements. One-way analysis of variance (ANOVA) and unpaired Student's t-test were used to compare the study groups. Post hoc Bonferroni test was used for the degree of significance between groups, and repeated measures analysis of covariance (ANCOVA) was used to investigate the effect of age and gender. IMA and MDA levels were significantly higher in the OCD group compared to the control group, and SOD, CAT and GSH-Px levels were lower in the OCD group compared to the control group (p<0.01). IMA levels were significantly higher in the OCD+MDD group compared to the OCD-MDD group, while SOD, CAT and GSH-Px levels were significantly lower in the OCD+MDD group compared to the OCD-MDD group (p<0101). MDA levels were significantly higher in the OCD+MDD group compared to the OCD-MDD group (p=0.009). When the entire OCD patient group was examined, significant, powerful, positive correlations were observed between Y-BOCS and HDRS scores and IMA and MDA, and significant powerful negative correlations between Y-BOCS and HDRS scores and SOD, CAT, and GSH-Px (p<0.001 for all). In OCD+MDD group, oxidative stress markers increased significantly in parallel with the severity of depression, while antioxidant levels decreased (p=0.003 for IMA, p<0.001 for others). We believe that parameters indicating impaired oxidant/antioxidant balance in patients with obsessive-compulsive disorder may help to elucidate the cause of the disease and may be potentially useful biomarkers in the diagnosis and determination of the severity of comorbid MDD.

Effects of mesenchymal stromal cells and human recombinant Nerve Growth Factor delivered by bioengineered human corneal lenticule on an innovative model of diabetic retinopathy.

Diabetic retinopathy (DR) is a microvascular complication of diabetes in which neurodegeneration has been recently identified as a driving force. In the last years, mesenchymal stromal cells (MSCs) and neurotrophins like Nerve Growth Factor (NGF), have garnered significant attention as innovative therapeutic approaches targeting DR-associated neurodegeneration. However, delivering neurotrophic factors directly in the eye remains a challenge. Hence, this study evaluated the effects of MSCs from human amniotic fluids (hAFSCs) and recombinant human NGF (rhNGF) delivered by human corneal lenticule (hCL) on a high glucose (HG) induced ex vivo model simulating the molecular mechanisms driving DR. Porcine neuroretinal explants exposed to HG (25 mM for four days) were used to mimic DR ex vivo. hCLs collected from donors undergoing refractive surgery were decellularized using 0.1% sodium dodecyl sulfate and then bioengineered with hAFSCs, microparticles loaded with rhNGF (rhNGF-PLGA-MPs), or both simultaneously. Immunofluorescence (IF) and scanning electron microscopy (SEM) analyses were performed to confirm the hCLs bioengineering process. To assess the effects of hAFSCs and rhNGF, bioengineered hCLs were co-cultured with HG-treated neuroretinal explants and following four days RT-PCR and cytokine array experiments for inflammatory, oxidative, apoptotic, angiogenic and retinal cells markers were performed. Data revealed that HG-treated neuroretinal explants exhibit a characteristic DR-phenotype, including increased level of NF-kB, NOS2, NRF2 GFAP, VEGFA, Bax/Bcl2 ratio and decreased expression of TUBB3 and Rho. Then, the feasibility to bioengineer decellularized hCLs with hAFSCs and rhNGF was demonstrated. Interestingly, co-culturing hAFSCs- and rhNGF- bioengineered hCLs with HG-treated neuroretinal explants for four days significantly reduced the expression of inflammatory, oxidative, apoptotic, angiogenic and increased retinal markers. Overall, we found for the first time that hAFSCs and rhNGF were able to modulate the molecular mechanisms involved in DR and that bioengineered hCLs represents a promising ocular drug delivery system of hAFSCs and rhNGF for eye diseases treatment. In addition, results demonstrated that porcine neuroretinal explants treated with HG is a useful model to reproduce ex vivo the DR pathophysiology.

6-Shogaol Abrogates Parkinson’s Disease in Rotenone-Induced Rodents: Based on In Silico Study and Inhibiting TNF-α/NF-κB/IL-1β/MAO-B

Background/Objectives: 6-Shogaol is a comparatively innovative anti-Parkinson’s remedy with antioxidant and anti-inflammatory characteristics. This investigation intended to determine the role of 6-shogaol in the Parkinson’s disease (PD) paradigm in rotenone-induced rats. Methods: Thirty male Wistar rats (10–12 weeks old; 180 ± 20 g) were divided into five groups. Animals with rotenone-induced experimental PD were subsequently treated with 6-shogaol-10 at 20 mg/kg for 28 days. After the experimental duration, behavioural investigations were performed, i.e., open field test, forced swim test, rotarod test, and catalepsy test. Biochemical assessments like AChE, GSH, CAT, SOD, MDA, nitrite, ceruloplasmin, proinflammatory markers such as IL-1β, NF-κB, TNF-α, and catecholamines markers (DA, GABA, and MAO-B) were determined. The docking procedure was conducted using the AutoDock Vina docking protocol. Furthermore, histopathology was performed. Results: Rotenone significantly increased the level of MAO-B, oxidative, nitrative, and pro-inflammatory markers. However, there was a decline in ceruloplasmin, dopamine, and endogenous antioxidants. Treatment with 6-shogaol (10 and 20 mg/kg) considerably sustained the elevation of oxidative stress and inflammatory indicators and decreased AChE activity and dopamine levels. In the histology of the brain, 6-shogaol improved the neuronal structure and reduced the degeneration of neurons. Based on the binding energy values, compound 6-shogaol demonstrates a favourable binding affinity to AChE, MAO-B, DA, and GABA with respective binding energies of −8.214, −8.133, −7.396 and −6.189 kcal/mol. Conclusions: In this study, 6-shogaol exhibited neuroprotective properties against PD, which could be employed as a prospective medication for PD.

Protective effect of evodiamine on acetic acid-induced gastric ulcers in rats through regulation of ROS/ICAM1/Nrf2 signaling pathway

Purpose: To study the effect of evodiamine on the signaling pathway of ROS/ICAM-1/Nrf2 in rats with acetic acid-induced stomach ulcers. Methods: The rats were randomly assigned to five groups containing 10 rats each. Prior to acetic acidinduction of gastric ulcers, omeprazole (4.0 mg/kg/day), low-dose evodiamine (L-EVD, 20 mg/kg/day), and high-dose evodiamine (H-EVD, 40 mg/kg/day) were orally administered to the respective groups for 15 days. Following ulcer induction, the same treatments continued for an additional 7 days, for a total treatment duration of 22 days. The control (0.9 % saline) and acetic acid (model) groups were administered 0.9 % sodium chloride solution (10 mL/kg) for the same period. Thereafter, oxidative stress, inflammatory markers, macroscopic and microscopic evaluations were conducted on the gastric mucosa. Results: The acetic acid group showed significantly higher levels of oxidative and inflammatory markers, as well as damage and degeneration of the gastric mucosa when compared to control group (p &lt; 0.05). However, both low-dose and high-dose evodiamine treatment groups demonstrated significant gastric healing. Administration of low-dose and high-dose evodiamine resulted in significantly lower levels of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), myeloperoxidase (MPO) and intracellular adhesion molecule-1 (ICAM-1, p &lt; 0.05). Furthermore, evodiamine treatment led to significantly increased levels of glutathione (GSH) and nuclear facterythroid factor 2 (Nrf2). Conclusion: Evodiamine reduces oxidative stress, suppresses inflammatory reactions, and exerts an anti-ulcer effect on acetic acid-induced gastric ulcers in rats by modulating ROS/ ICAM-1/Nrf2 signaling pathway. More studies into the integration of evodiamine into conventional pharmaceutical treatments for gastric ulcers should be conducted.

Role of oxidative stress in the occurrence of periodontitis in the background of metabolically associated diseases

Oxidative stress is an important component of slow pathological inflammation in metabolic diseases and chronic periodontitis. Periodontal diseases are interrelated with somatic pathology. This results in both local and systemic inflammatory reactions, which increases the risk of the onset and progression of somatic diseases. Psychological stress associated with oxidative stress with increased production of oxidants and oxidative damage is an important factor in the etiology of both periodontitis and metabolic disorders. Objective — to analyze the possible bond between chronic periodontitis, 8‑hydroxydeoxyguanosine (8‑OHdG), as a biomarker of oxidative stress, and cortisol in persons with metabolic‑associated diseases. Materials and methods. 68 people were surveyed, of which 47 had metabolically ‑associated diseases that were diagnosed according to domestic and international criteria. The control group consisted of systematically healthy people, representative by age and article that did not receive dental therapeutic and preventive measures for the last 6 months. The diagnosis of «chronic periodontitis» was carried out in accordance with existing recommendations provided at World Workshop on the Classification of Periodontal and Peri­Implant Diseases and Conditions (2017). The concentration of 8‑OHdG in plasma and the level of saliva cortisol were measured by the immunoenzyme method according to the manufacturer’s instructions. The arithmetic mean, standard error, and Student’s t‑test were calculated to compare the characteristics. Pearson’s correlation test was used to identify the relationship between the indicators. Results. Each of the groups had patients who had a healthy periodontium and those who had chronic periodontitis. The 8‑OHdG level in the main group was much higher (p&lt;0.001) than in the control group, and the difference in the concentration of 8‑OHdG was also observed between those patients who had periodontal disease and those who had a healthy periodontium. The level of salivary cortisol in patients with chronic periodontitis was much higher compared to patients with clinically healthy periodontium (p=0.00). A correlation was found between cortisol levels and the presence of periodontal disease (r=0.619; p=0.00), between plasma 8OHdG levels and the epithelial attachment loss index (r=0.537; p=0.00), the hygiene index (r=0.599; p=0.00), and between 8OHdG levels and salivary cortisol concentrations (r=0.292; p=0.036). Conclusions. Oxidative stress plays an important role in delayed pathological inflammation in both metabolic diseases and chronic periodontitis. 8‑OHdG is a key and universal biomarker for the evaluation of DNA oxidative damage. There is a positive relationship between the oxidative marker, periodontitis and cortisol.

Evidence That Long-Term Treatment Prevents Tissue Oxidative Damage in Patients With Inherited Disorders of the Propionate Pathway.

Propionic and methylmalonic acidemias (PAcidemia and MMAcidemia, respectively) are genetic disorders clinically characterized by metabolic decompensation associated with life-threatening encephalopathic episodes in the neonatal period. Adequate and rapid therapeutic management is essential for patients' survival and prognosis. In this study, a restricted protein diet associated with L-carnitine (LC) supplementation was shown to decrease mortality and morbidity in patients affected by these disorders probably by decreasing the accumulation of the major metabolites and therefore their toxicity. Since oxidative stress was proposed as a contributing mechanism of tissue damage in PAcidemia and MMAcidemia and LC has potent antioxidant properties, our objective in this work was to investigate the effects of a long-term therapy consisting of reduced protein intake associated with LC supplementation on oxidative damage markers in patients affected by these diseases. We measured urinary isoprostanes, di-tyrosine, and oxidized guanine species, which reflect oxidative damage to lipids, proteins, and DNA/RNA, respectively, as well as the concentrations of NO products (nitrate plus nitrite) in patients untreated or submitted to short-term or a long-term treatment. Results revealed significant increases of isoprostanes, di-tyrosine, and oxidized guanine species, as well as a moderate nonsignificant increase of NO levels in the untreated patients, relatively to controls. Furthermore, these altered markers were attenuated after short-term treatment and normalized after prolonged treatment. In conclusion, data from this work show for the first time that long-standing treatment of patients with disorders of the propionate pathway can protect against oxidative damage. However, it remains to be elucidated whether oxidative stress identified in this study directly correlates with the clinical conditions of the affected patients.

Taxus chinensis (Pilg.) Rehder fruit attenuates aging behaviors and neuroinflammation by inhibiting microglia activation via TLR4/NF-κB/NLRP3 pathway

Ethnopharmacological relevanceAs one of the important by-products of Taxus chinensis (Pilg.) Rehder, its fruit (TCF) has a sweet taste, which is commonly used in folklore to make health care wine reputed for enhancing immune function and promoting anti-aging effects, especially popular in the longevity villages of China for a long history. Evidences had showed that Taxus chinensis fruit contained polysaccharides, flavonoids, amino acids and terpenoids, which all were free of toxic compounds , but its medicinal value has not been fully recognized. Our previous studies have found that TCF extract may reverse many biological events, including oxidative stress, inflammatory response, neuronal apoptosis, etc. by in silico methods, suggesting potential avenues for future pharmaceutical exploration in aging and age-related diseases. Aim of the studyYet, the anti-aging properties of TCF have not been specifically studied, this study aims to fill this gap by investigating the effects of TCF extract (TCFE) in an aging mouse model, particularly focusing on its role in inhibiting microglial activation and elucidating its underlying anti-aging mechanisms. Materials and methodsAn aging mouse model was induced using D-galactose, with interventions involving high, medium, and low doses of TCFE compared to a positive control (2mg/kg rapamycin combined with 100mg/kg metformin). The methodology involved evaluating behavioral changes, serum oxidative and antioxidative markers, hypothalamic β-galactosidase activity, expression of the aging-related protein P63, serum inflammatory factors, and the TLR4/NF-κB/NLRP3 inflammatory pathway in hypothalamic tissues. Additionally, to strengthen our in vivo findings, we conducted in vitro experiments on LPS-stimulated BV2 microglial cells. Finally, UPLC-MS/MS for precise component analysis using compound standards, coupled with molecular docking analyses, were employed to discern and elucidate the anti-inflammatory mechanisms of TCF. ResultsIn vivo results revealed TCFE significantly ameliorated behavioral deficits, reduced oxidative stress markers (MDA) and pro-inflammatory cytokines (IL1-β, IL-6, IFNg, TNFα, IL-17), and increased in antioxidants (SOD, T-AOC) and anti-inflammatory factors (IL-10). TCFE also reduced hypothalamic senescence, improved cellular integrity, lowered p63, and inhibited microglia activation and inflammatory pathways (TLR4, NFKB, NLRP3). The overall effect of TCFE was better than that of the positive drug group (rapamycin combined with metformin). In vitro results further revealed that TCFE markedly decreased IL1-β, NFKB, and TLR4 levels in BV2 microglial cells, showing comparable efficacy to a TLR4 classic positive inhibitor C34, supporting its anti-inflammatory role. Through UPLC-MS/MS analysis coupled with compound standards, we identified ten bioactive compounds, including gallocatechin, epigallocatechin, catechin, procyanidin B2, kaempferol, quercetin, rutin, naringin, apigenin, ginkgetin. All these compounds showed strong binding affinity to TLR4, notably procyanidin B2 and rutin, potentially through hydrogen bonds, aromatic cation-π interactions, and hydrophobic interactions, suggesting a molecular basis for their anti-inflammatory action. ConclusionTCFE showed strong anti-aging effects by inhibiting microglia activation and lessening oxidative stress and modulating inflammatory pathways. This research supports TCF's use in anti-aging and sets a base for future drug development in the realms of neuroinflammation and aging.