Oxidative Stress Gene Expression Research Articles

Yeast Rhodoturula glutinis as a modulator of innate immune and oxidative stress-related genes in Oreochromis niloticus cultured in a Biofloc system

The effect of live yeast Rhodoturula glutinis was evaluated on juvenile Oreochromis niloticus cultivated in a Biofloc system. Growth performance and the expression of innate response and relevant oxidative stress genes were evaluated after a 12-week feeding trial. Three experimental treatments were evaluated in a conventional tilapia culture (control), Biofloc culture (BFT), and Biofloc with the addition of the live yeast R. glutinis 1×106 CFU g-1 (BFT+Rg), with four replicates per treatment. In all cases, commercial food was supplied to the organisms (32% protein and 5% lipids). O. niloticus juveniles (7.02 ± 0.04 g) were randomly distributed in 12 tanks, each with 15 animals. BFT+Rg treatment showed a significant increase in weight gain compared with the other treatments. Significant improvements were found in Fulton's condition factor, feed conversion rate, and hepatosomatic index under Biofloc conditions, BFT, and BFT+Rg treatments. No significant differences were observed in survival. R. glutinis directly influenced gene expression in the liver and intestine. The expression of tnfa, tgfb, hsp70, and gpx, genes in the liver significantly increased in the BFT+Rg treatment compared with the other treatments. Similarly, a significant increase was found in intestinal il1b, tnfa, tgfb, trf, hsp70 gpx, and cat expression patterns in the BFT+Rg treatment. Based on the performance and immune response, the present study suggests the use of R. glutinis as a strategy to increase the productivity of tilapia in Biofloc culture.

Efficacy of chlorogenic acid against ethylene glycol-induced renal stone model: The role of NFKB-RUNX2-AP1-OSTERIX signaling pathway

Background and objectiveRenal tissue injuries by free radicals are an essential reason in pathogenesis of urinary tract stones. Ethylene glycol is one of the toxic agents which can causes to the increases in biosynthesis of reactive oxygen species and oxidative stress condition. Natural antioxidants have been reported to protective efficacy against renal stones formation. Accordingly, the aim of the current experiment was to identify the renal protective effect of chlorogenic acid as a well-prominent antioxidant on ethylene glycol-induced renal stone model targeting the NFKB-RUNX2-AP1-OSTERIX signaling pathway. Materials and methodsRenal stones model were established by ethylene glycol (Percent: 0.75) within the daily drinking water for rats. Treatment groups received cystone (750 mg/kg) and chlorogenic acid (100, 200, and 400 mg/kg, day: 15th to 28th, gavage). After 4 weeks, the renal function parameters (calcium, uric acid, creatinine, total protein, oxalate, and citrate) in plasma, urine, and renal tissue were measured. Moreover, oxidative stress factors and gene expression of NFKB, RUNX2, AP1, and OSTERIX were also evaluated. ResultsThe results showed improved renal function in chlorogenic acid-treated groups. The total proteins and creatinine excretion were decreased. Also the gene expression of oxidative stress pathway (NFKB-RUNX2-AP1-OSTERIX) were decreased which caused to increases of antioxidant enzymes. Conclusionsthe antioxidant activity increases by chlorogenic acid treatment may have a critical role in prevention of calcium oxalate formation via inhibition of the NFKB-RUNX2-AP1-OSTERIX signaling pathway.

Resveratrol Mitigates Oxygen and Glucose Deprivation-Induced Inflammation, NLRP3 Inflammasome, and Oxidative Stress in 3D Neuronal Culture.

Oxygen glucose deprivation (OGD) can produce hypoxia-induced neurotoxicity and is a mature in vitro model of hypoxic cell damage. Activated AMP-activated protein kinase (AMPK) regulates a downstream pathway that substantially increases bioenergy production, which may be a key player in physiological energy and has also been shown to play a role in regulating neuroprotective processes. Resveratrol is an effective activator of AMPK, indicating that it may have therapeutic potential as a neuroprotective agent. However, the mechanism by which resveratrol achieves these beneficial effects in SH-SY5Y cells exposed to OGD-induced inflammation and oxidative stress in a 3D gelatin scaffold remains unclear. Therefore, in the present study, we investigated the effect of resveratrol in 3D gelatin scaffold cells to understand its neuroprotective effects on NF-κB signaling, NLRP3 inflammasome, and oxidative stress under OGD conditions. Here, we show that resveratrol improves the expression levels of cell viability, inflammatory cytokines (TNF-α, IL-1β, and IL-18), NF-κB signaling, and NLRP3 inflammasome, that OGD increases. In addition, resveratrol rescued oxidative stress, nuclear factor-erythroid 2 related factor 2 (Nrf2), and Nrf2 downstream antioxidant target genes (e.g., SOD, Gpx GSH, catalase, and HO-1). Treatment with resveratrol can significantly normalize OGD-induced changes in SH-SY5Y cell inflammation, oxidative stress, and oxidative defense gene expression; however, these resveratrol protective effects are affected by AMPK antagonists (Compounds C) blocking. These findings improve our understanding of the mechanism of the AMPK-dependent protective effect of resveratrol under 3D OGD-induced inflammation and oxidative stress-mediated cerebral ischemic stroke conditions.

PSXI-6 Effects of a Saccharomyces Cerevisiae Fermentation Product-Supplemented Diet on Circulating Immune Cells and Oxidative Stress Markers of Dogs

Abstract Feeding Saccharomyces cerevisiae fermentation product (SCFP) has previously altered fecal microbiota, fecal metabolites, and immune cell function of adult dogs. The objective of this study was to further measure circulating immune cell numbers and activity, antioxidant status, and oxidative stress marker concentrations of adult dogs fed SCFP in an extruded diet. Sixteen adult pointer dogs (8M, 8F; mean age = 8.35 y; mean BW = 27.13 kg) were used in a randomized crossover design study. All dogs were fed a control diet for 4 wk, then randomly assigned to the control or SCFP-supplemented diet and fed to maintain BW for 10 wk. A 6-wk washout preceded the second 10-wk experimental period with dogs receiving opposite treatments. After baseline/washout and treatment phases, pre- and post-prandial blood samples were collected. Oxidative stress and immune cell function were measured using commercial ELISA kits, circulating immune cell numbers were analyzed using flow cytometry, and mRNA expression of oxidative stress genes were analyzed using RT-PCR. Change from baseline data were analyzed using the Mixed Models procedure of SAS 9.4, with P< 0.05 being significant and P< 0.10 being trends. Changes in unstimulated lymphocytes and stimulated IFN-g secreting T-cells were lower (P< 0.05) in SCFP-fed dogs, while change in stimulated T cells were lower (P< 0.05) in control-fed dogs. The stimulated cytotoxic T cells delta trended lower (P< 0.10) in SCFP-fed dogs. Change in serum superoxide dismutase (SOD) concentrations were higher (P< 0.05) and change in catalase mRNA expression was lower (P< 0.05) in SCFP-fed dogs. All other immune cells, oxidative stress markers, and gene expression were unaffected by treatment. In conclusion, SCFP-fed dogs have increased SOD, which supports lower acute-inflammatory responses and aligns with some immune stimulation data.

123 Effects of a Saccharomyces Cerevisiae Fermentation Product-Supplemented Diet on Fecal Characteristics, Oxidative Stress, and Blood Gene Expression of Adult Dogs Undergoing Transport Stress

Abstract Previously, a Saccharomyces cerevisiae fermentation product (SCFP) was shown to positively alter fecal microbiota, fecal metabolites, and circulating immune cell function of adult dogs. The objective of this study was to measure the effects of SCFP on fecal characteristics, serum oxidative stress biomarkers, and whole blood gene expression of adult dogs undergoing transport stress. Sixteen adult pointer dogs (8M, 8F; mean age = 8.35 y; mean BW = 27.13 kg) were used in a randomized crossover design study. All dogs were fed control diet for 4 wk, then randomly assigned to a control or SCFP-supplemented diet and fed to maintain BW for 11 wk. A 6-wk washout preceded the second 11-wk experimental period with dogs receiving opposite treatments. After 11 wk, fresh fecal and blood samples were collected before and after transport in a van for 45 min. Change from baseline data (i.e., before and after transport) were analyzed using the Mixed Models procedure of SAS 9.4, with P< 0.05 being significant and P< 0.10 being trends. Changes in serum malondialdehyde and 8-isoprostane:creatinine concentrations increased (P< 0.05) in dogs fed SCFP, but decreased in control dogs with transport stress. Other serum markers were unaffected by diet during transport stress. Fecal dry matter percentage tended to be affected (P< 0.10) by diet during transport stress, being reduced in control dogs, but stable in dogs fed SCFP. Other fecal characteristics were unaffected by diet during transport stress. Blood cyclooxgenase-2 and myeloperoxidase mRNA expression was affected (P< 0.05) by diet during transport stress, being increased in control dogs, but stable or decreased in dogs fed SCFP. Expression of other genes was unaffected by diet during transport stress. These data suggest that the benefits of feeding a SCFP in a novel transport stress model are mediated more so by suppressing innate immune cell activation than through decreasing lipid peroxidation.

Antimony accumulation in zebrafish (Danio rerio) and its effect on genotoxicity, histopathology, and ultrastructure

Antimony (Sb) is a toxic metal in aquatic ecosystems. In this study, the accumulation of aqueous Sb in the liver, brain, gills and muscle of zebrafish (Danio rerio) and its effect on genotoxicity, histopathology and ultrastructure alterations were evaluated. The fishes were exposed to different concentrations (0, 8.29, 16.58, 33.16 mg L−1) of aqueous Sb for 18 days. The results showed that the order of Sb accumulation in different tissues was liver > gill > muscle > brain, and the accumulation increased with increasing Sb stress concentration. The mRNA expression levels of Nrf2, Cu/Zn-SOD, Mn-SOD, CAT and GPx genes showed different trends. In addition, significant histopathology and ultrastructure alterations were observed in the liver and gills exposed to Sb. Sb could accumulate in different tissues of zebrafish, inducing the expression of oxidative stress genes and activating antioxidant defense systems. Histological and ultrastructural changes could be used as valid biomarkers for the assessment of aqueous Sb contamination.

Neurotoxicity associated with oxidative stress and inflammasome gene expression induced by allethrin in SH-SY5Y cells.

Pyrethroids, including allethrin, have largely been used as commercial insecticides. The toxicity of allethrin is little known, but it is assumed that, as occurs with other pyrethroids, it could cause alterations of the nervous system and pose both occupational and non-occupational health hazards. To evaluate the neurotoxicity of allethrin we used the MTT assay of SH-SY5Y neuroblastoma cells to determine cell viability. Dose-dependent reductions of cell viability served to compare the vehicle-group and the IC50 for allethrin, which was 49.19μM. ROS production increased significantly at concentrations of 10-200μM of allethrin, and NO levels were significantly increased by the effect of allethrin at a minimum concentration of 50μM. Lipid peroxidation increased by the effect of allethrin at concentrations of 25, 50, 100, and 200μM. Caspase 3/7 activity was induced by allethrin concentrations of 50, 100, and 200μM. Here, we suggest that allethrin might affect the inflammasome complex (Caspase-1, NLRP3, and PYDC1) and apoptosis (Bax and Bcl-2) gene expression by mRNA fold change expression levels shown in Caspase-1 (2.46-fold), NLRP3 (1.57-fold), PYDC1 (1.48-fold), and Bax (2.1-fold). These results demonstrated that allethrin induced neurotoxicity effects on SH-SY5Y cells through activation of inflammasome pathways, cell death, and oxidative stress.

Effects of a Saccharomyces cerevisiae fermentation product-supplemented diet on circulating immune cells and oxidative stress markers of dogs.

Feeding Saccharomyces cerevisiae fermentation product (SCFP) has previously altered fecal microbiota, fecal metabolites, and immune function of adult dogs. The objective of this study was to investigate measures of skin and coat health, changes in circulating immune cell numbers and activity, antioxidant status, and oxidative stress marker concentrations of healthy adult dogs fed a SCFP-supplemented extruded diet. Sixteen adult English Pointer dogs (8 M, 8 F; mean age = 6.7 ± 2.1 yr; mean BW = 25.9 ± 4.5 kg) were used in a randomized crossover design study. All dogs were fed a control diet for 4 wk, then randomly assigned to either the control or SCFP-supplemented diet (0.13% of active SCFP) and fed to maintain BW for 10 wk. A 6-wk washout preceded the second 10-wk experimental period with dogs receiving opposite treatments. After baseline/washout and treatment phases, skin and coat were scored, and pre and postprandial blood samples were collected. Transepidermal water loss (TEWL), hydration status, and sebum concentrations were measured (back, inguinal, ear) using external probes. Oxidative stress and immune cell function were measured by ELISA, circulating immune cell percentages were analyzed by flow cytometry, and mRNA expression of oxidative stress genes was analyzed by RT-PCR. Change from baseline data was analyzed using the Mixed Models procedure of SAS 9.4. Sebum concentration changes tended to be higher (P < 0.10; inguinal, ear) in SCFP-fed dogs than in controls. TEWL change was lower (P < 0.05) on the back of controls, but lower (P = 0.054) on the ear of SCFP-fed dogs. Delayed-type hypersensitivity response was affected by diet and time post-inoculation. Other skin and coat measures and scores were not affected by diet. Changes in unstimulated lymphocytes and stimulated IFN-γ secreting T cells were lower (P < 0.05) in SCFP-fed dogs, while changes in stimulated T cells were lower (P < 0.05) in control-fed dogs. Upon stimulation, the percentage of cytotoxic T cells delta trended lower (P < 0.10) in SCFP-fed dogs. Change in serum superoxide dismutase concentrations was higher (P < 0.05) and change in catalase mRNA expression was lower (P < 0.05) in SCFP-fed dogs. All other measurements of immune cell populations, oxidative stress markers, and gene expression were unaffected by treatment. In conclusion, our data suggest that SCFP positively impacts indicators of skin and coat health of dogs, modulates immune responses, and enhances some antioxidant defense markers.

Effects of Lactobacillus kefiri fermentation supernatant on skin aging caused by oxidative stress

• • Lactobacillus kefiri fermentation components(LAF) shows excellent antioxidant effect in vitro. • •Achievements of Lactobacillus kefiri fermentation components providing more possibilities for their application as functional raw materials in cosmetics. • •LAF can regulate the expression of oxidative stress, autophagy, and aging-related genes in damaged cells treated with H 2 O 2 , thereby delaying cell senescence. Lactobacillus kefiri is a beneficial bacteria that is often used to make yogurt only, and is beneficial to human health when it is used in food fermentation. In this article, a fermentation supernatant (LAF) is prepared by culturing Lactobacillus kefiri , and the anti-aging effects of LAF are evaluated at the biochemical, cellular, and molecular biology levels. The antioxidant activity of LAF is comprehensively evaluated by three free radical scavenging tests and the total antioxidant capacity (ABTS method) test. The toxicity of LAF on human skin fibroblasts (HSF) is detected using the MTT method, and the effects of LAF on H 2 O 2 -induced oxidative stress in terms of intracellular reactive oxygen species (ROS), Malondialdehyde (MDA) content, antioxidant enzyme activity are evaluated. The gene expression levels of LC3 and Beclin-1 are detected using the real-time reverse transcription-polymerase chain reaction (qRT-PCR) method, and the protein expression levels of LC3 are detected using the Western blot method to evaluate the effects of LAF on HSF autophagy. The expression of type I collagen (COL-1), Matrix metalloproteinase-1(MMP-1), AKT pathway, MAPK pathway, and other genes are detected by qRT-PCR to study the anti-aging effects of LAF. From the experimental results, it is found that LAF can delay the aging of HSF at the biochemical, cellular, and molecular biology levels, and can be utilized as a functional food ingredient.

Ameliorating Effects of Bioactive Peptides Extracted from Litopenaeus vannamei Wastes on Oxidative Stress, Glucose Regulation, and Autophagy Gene Expression in Nonalcoholic Fatty Liver-Induced Rats.

This study aimed to investigate the effect of bioactive peptides from Litopenaeus vannamei on oxidative stress, glucose regulation, and autophagy gene expression in the induced nonalcoholic fatty liver rats. Bioactive peptides used in the current study were extracted in a progressive rise in temperature (40-60°C) (GP). For this purpose, twenty-four healthy male rats (initial weight, 230.1 ± 22 g) were divided in four experimental groups including control (standard diet), HFD (high-fat diet), HFD + GP20, and 300 (high-fat diet + 20, 300 mg peptides/kg body weight). After 70 days, the results indicated that experimental treatments did not affect the body and liver weight (P > 0.05), although the higher liver weight was seen in HFD treatment. Based on these results, the use of GP peptides improved antioxidant enzymes and decreased MDA concentration, and a significant difference was observed between peptide treatments and HFD (P < 0.05). In comparison to the HFD group, significantly lower liver enzymes (ALT and AST) were seen in peptide treatments (P < 0.05). Also, the results indicated that the lowest amylase, alkaline phosphatase, glucose, insulin, HOMA-IR, and inflammation cytokines (TNF-ɑ and IL-6) were seen in peptide groups. The autophagy gene expression was measured in the liver cells, and the results showed that, unlike HFD treatment, the use of GP peptides decreased Beclin 1, Atg7, and P62 expression in male rat's livers. Overall, the results of the current study demonstrated that the use of GP peptides at low concentration shows significant hypoglycemia and antioxidant properties in nonalcoholic fatty liver-induced rats.

Transcriptional modulation of inflammation, and aging in Indian obese adults following a 12-week yoga-based lifestyle intervention: A randomized controlled trial.

IntroductionObesity is one of the major global problems in today's world, both in children, and the adult age group. Current evidence suggests obesity alters the expression of various genes related to oxidative stress, inflammation, and aging. In recent times complementary therapy like yoga-based lifestyle intervention (YBLI) is used as an adjunct therapy to modern medicine. This study examines the efficacy of 12 weeks of yoga-based lifestyle intervention with standard care (SC) on the expression of genes related to oxidative stress, inflammation, and aging in obese adults.MethodsThis was a two-arm parallel randomized control trial implemented at Integral Health Clinic (IHC), an outpatient facility that regularly conducted YBLI programs for the prevention of lifestyle diseases like obesity and diabetes in the Department of Physiology, All India Institute of Medical Sciences (AIIMS), New Delhi. Blood samples at baseline and weeks 2,4, and 12 were collected from 72 adults (male n = 21; female n = 51) of age 20–45 years with a body–mass index (BMI) of 25–35 kg/m2 who were randomized to receive either a 12-week SC (n = 36) or YBLI (n = 36). SC included recommendations for the management of obesity as per Indian guidelines including a low-calorie individualized diet and physical activity. Asana (physical postures), pranayama (breathing exercises), and meditation were all part of the YBLI. Primary outcomes were relative fold change in the expression of genes associated with oxidative stress [Nuclear factor-kappa B (NF-Kappa B)], inflammation [Tumor necrosis factor-α (TNFα), interleukin-6 (IL-6)], and aging [human telomerase reverse transcriptase (TERT)] in peripheral blood mononuclear cells between the two groups at week-12.ResultsThere were no significant changes in fold change of TERT, IL-6, and NF-kappa B between the groups at week 12. The relative fold change of TERT was significantly greater in the YBLI group (p = <0.0001) vs the SC group at 2 weeks. The relative fold change of TNF α was significantly lower at week 12 in YBLI though the change was not continuous and reliable. Within both groups, TERT expression was significantly increased at week 2 though the change was greater in the YBLI group (p < 0.0001). TNF α gene expression was significantly lower at weeks 2 and 4, compared to baseline level, in the SC group but it increased at week 12.ConclusionThe results while did not confirm our hypothesis, are important to share with the scientific society, to be able to improve prospective study designs and find optimal time/intervention/biological marker settings for this highly important scientific field. The results are suggestive of a positive impact of YBLI and SC on the fold change of aging-related TERT gene in obesity, though the benefit was not evident till week 12. However, the results should be evaluated with caution and in light of other published studies. To better understand the positive effects of YBLI on oxidative stress, inflammation, and aging-related gene expression in obesity, larger studies are recommended.

Resveratrol treatment modulates several antioxidant and anti-inflammatory genes expression and ameliorated oxidative stress mediated fibrosis in the kidneys of high-fat diet-fed rats

ObjectiveResveratrol is a polyphenolic compound that possesses strong antioxidant and anti-inflammatory activities. This study evaluated the effects of resveratrol on oxidative stress, fibrosis and multiple genes regulation in the kidneys of high fat (HF) diet-fed rats. MethodsWistar rats were fed with HF diet for eight weeks. These rats were also treated with resveratrol for eight weeks. Finally, kidney tissue samples were isolated from all sacrificed rats. The histological changes, creatinine and uric acid levels, oxidative stress parameters such as malondialdehyde (MDA), nitric oxide, and advanced oxidation protein product (AOPP) levels were analyzed. The antioxidant enzymes such as catalase, superoxide dismutase (SOD) activities and reduced glutathione (GSH) levels; gene expression of inflammatory and fibrosis-related genes namely, inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), transforming growth factor beta1 (TGF-β1), and collagen-1 were assessed. Moreover, gene expression of oxidative stress-related genes such as nuclear factor erythroid 2–related factor 2(Nrf-2), SOD, catalase, and glutathione reductase, were also assessed. ResultsHF diet-fed rats showed increased creatinine and uric acid levels in plasma which were lowered by resveratrol treatment. The study findings also revealed that resveratrol counterbalanced the oxidative stress and prevented the expression of the inflammatory genes; restored the catalase and SOD activities followed by the up-regulation of antioxidant genes expression in the kidneys of HF diet-fed rats. HF diet caused the Nrf-2 down-regulation followed by the decreased expression of HO-1 and HO-2 genes, which was restored by resveratrol treatment. Moreover, the histological assessment showed lipotoxicity and increased fibrosis in the kidneys of HF diet-fed rats. Resveratrol prevented the kidney fibrosis probably by limiting oxidative stress, inflammation, and down-regulating TGF-β1 mediated signaling pathway. ConclusionIn conclusion, resveratrol treatment showed beneficial effects in preventing oxidative stress and fibrosis in the kidneys of HF diet-fed rats probably by modulating the gene expression of oxidative stress and inflammation related factors and enzymes.

Synergistic gentamicin-photodynamic therapy against resistant bacteria in burn wound infections

BackgroundMulti-resistant bacteria, partially a result of the abuse of antibiotics, have greatly diminished the effectiveness of antibiotics. The combination of antibiotics with other therapies like antimicrobial photodynamic therapy (aPDT) may provide a useful strategy for fighting resistant bacteria. Here, the synergistic bactericidal effects of toluidine blue (TB)-aPDT and gentamicin (GEN) were evaluated in vitro and in vivo. MethodsThe Post-antibacterial effects were measured at 600 nm (OD600) by a microplate reader. The bacterial envelope and biofilm were observed by a field emission scanning electron microscope. The expression of oxidative stress and Agr system-related genes was analyzed by qRT-PCR after GEN combined with TB-aPDT (GEN&aPDT). Besides, the burn infection model was established to investigate the cloning efficiency of immobilized bacteria, wound healing and inflammatory factors in the lesions. ResultsGEN&aPDT could inhibit the growth of Staphylococcus aureus (S. aureus) and multidrug-resistant S. aureus (MDR S. aureus) for up to 15 h, and destroyed the cell envelope and biofilm structure of S. aureus and MDR S. aureus. During the process, ROS played an important role, inducing oxidative stress and downregulating the expression of AgrA, AgrB and PSM in the Agr system, resulting in decreased bacterial virulence and infectivity. In addition, GEN&aPDT cotreatment could effectively promoted wound healing in burn-infected mice by reducing the numbers of bacterial colonization in the wound, decreasing the content of inflammatory factors, and increasing the expression of growth factors. ConclusionThe present study confirmed a bactericidal synergy between GEN and aPDT in vitro and in vivo, therein, the oxidative stress exhibited an important role in decreasing bacterial virulence and infectivity, which may bring new ideas for the treatment of bacterial resistance.

Orlistat, a competitive lipase inhibitor used as an antiobesity remedy, enhances inflammatory reactions in the intestine

Alterations in secondary gut metabolites derived from the microbial fermentation of food in the gut have significant effects on various aspects of host physiology. Our recent studies on obese mice treated with Orlistat, an antiobesity treatment, revealed a significantly altered gut microbial profile marked by an over-abundance of Proteobacteria and alterations in secondary gut metabolites. In this study, we determined effect of fecal metabolites from high-fat diet fed mice treated with Orlistat (HFDOrl) on colonic epithelial cells in relation to inflammation, barrier function, mitochondrial activity, reactive oxygen species (ROS) levels, and oxidative stress. Quantitative PCR was used to measure intestinal mRNA expression of oxidative stress, inflammation, apoptosis, and gut barrier function genes in mice on a high fat diet with and without Orlistat treatment versus those fed a low-fat diet (HFDOrl, HFD, Normal diet-fed [ND] respectively). Alterations to antioxidant function in HCT-116-ARE-luciferase stable cell line and mitochondrial function in Caco-2 cells was analyzed under oxidative stress with exposure to aqueous fecal extracts from HFDOrl, HFD, and ND groups. The results of this study indicate that a significant increase in anti-oxidative response was observed based on the luciferase activity of HCT-116-ARE-luciferase stable cells. Increased maximal respiration and mitochondrial ROS under oxidative stress was also detected in confluent Caco-2 cells resulting from exposure to fecal extracts from the HFDOrl group compared with the HFD group and pure Orlistat. Furthermore, mice from the HFDOrl group exhibited a significant increase in colonic epithelial expression of oxidative markers (Nrf-2 and SOD-2), inflammation-related markers (IL-6 and TNF-α), and gut barrier function markers (Muc-2 and Occludin). Taken together, the results suggest that Orlistat treatment in the HFD group causes changes in secondary gut metabolites which affect the colonic redox state and may eventually lead to the development of inflammatory, oxidative, and mitochondrial dysfunction at the cellular level.

Concurrent alteration in inflammatory biomarker gene expression and oxidative stress: how aerobic training and vitamin D improve T2DM

BackgroundVitamin D (Vit D) supplementation and Aerobic Training (AT) exert several beneficial effects such as antioxidant and anti-inflammatory actions. The literature on the effects of AT and Vit D supplementation on the oxidative stress biomarkers and gene expression of inflammatory cytokines in patients with Type 2 Diabetes Mellitus (T2DM) is limited. The present study aimed to examine the effects of AT and Vit D supplementation on inflammation and oxidative stress signaling pathways in T2DM patients.Materials and methodsIn this single-blinded, randomized, placebo-controlled trial, 48 men with T2DM (aged 35–50 years with Body Mass Index (BMI) of 25–30 kg/m2) were randomly allocated into four groups: AT+Vit D (n = 10); AT + placebo (AT; n = 10); Vit D (n = 10), and Control + placebo (C; n = 10). The eight-week AT program was executed for 20–40 min/day, at 60–75% of heart rate maximum (HRmax), for 3 days/wks. The Vit D group received 50,000 IU of Vit D supplement capsules per week for 8 weeks. The serum levels of oxidative stress biomarkers and gene expression of inflammatory cytokines in the Peripheral Blood Mononuclear Cells (PBMCs) were evaluated using the RT-PCR method. To analyze the data, paired t-tests and one-way analysis of variance and Tukey’s post hoc test were used at the significance level of P < 0.05.ResultsThe result shows that serum 25-OH-Vit D, total nitrite, Total Glutathione (GSH), Total Antioxidant Capacity (TAC), Superoxide Dismutase (SOD), Catalase (CAT), and Glutathione Peroxidase (GPX) increased; and insulin, Fasting Blood Glucose (FBG), Homeostasis Model Assessment of Insulin Resistance (HOMA-IR), High Sensitivity C-Reactive Protein (hs-CRP), Malondialdehyde (MDA), glycated albumin, and Urinary 8-hydroxydeoxyguanine (8-OHdG) decreased significantly in all groups after 8 weeks, except for C. In addition, results of RT-PCR showed that AT+Vit D, Vit D, and AT significantly downregulated the gene expression of Tumor Necrosis Factor-Alpha (TNF-α), Interleukin-1 Beta (IL-1β), Mitogen-Activated Protein Kinases 1 (MAPK1), Nuclear Factor Kappa B (NF-κB) 1 (p50). It also upregulated Interleukin-4 (IL-4) gene expression, Peroxisome Proliferator-Activated Receptor Gamma (PPAR-γ) in T2DM patients compared to the C.ConclusionAdditionally, the AT+Vit D group showed significantly lower insulin, FBG, HOMA-IR, hs-CRP, MDA, glycated albumin, urinary 8-OHdG, IL-1β, TNF-α, MAPK1, and NF-κB1 (p50) levels and significantly higher serum 25-OH-Vit D, total nitrite, GSH, TAC, CAT, SOD, GPX, IL-4, and PPAR-γ levels compared to the AT and Vit D groups. In T2DM patients, 8 weeks of AT+Vit D had a more significant impact on certain gene expressions related to inflammation and oxidative stress than Vit D or AT alone.

Effect of sulfamethazine on the horizontal transfer of plasmid-mediated antibiotic resistance genes and its mechanism of action

As a new type of environmental pollutant, antibiotic resistance genes (ARGs) pose a huge challenge to global health. Horizontal gene transfer (HGT) represents an important route for the spread of ARGs. The widespread use of sulfamethazine (SM2) as a broad-spectrum bacteriostatic agent leads to high residual levels in the environment, thereby increasing the spread of ARGs. Therefore, we chose to study the effect of SM2 on the HGT of ARGs mediated by plasmid RP4 from Escherichia coli (E. coli) HB101 to E. coli NK5449 as well as its mechanism of action. The results showed that compared with the control group, SM2 at concentrations of 10 mg/L and 200 mg/L promoted the HGT of ARGs, but transfer frequency decreased at concentrations of 100 mg/L and 500 mg/L. The transfer frequency at 200 mg/L was 3.04 × 10−5, which was 1.34-fold of the control group. The mechanism of SM2 improving conjugation transfer is via enhancement of the mRNA expression of conjugation genes (trbBP, trfAP) and oxidative stress genes, inhibition of the mRNA expression of vertical transfer genes, up regulation of the outer membrane protein genes (ompC, ompA), promotion of the formation of cell pores, and improvement of the permeability of cell membrane to promote the conjugation transfer of plasmid RP4. The results of this study provide theoretical support for studying the spread of ARGs in the environment.

Silica Nanoparticle–Induced Reproductive Toxicity in Male Albino Rats via Testicular Apoptosis and Oxidative Stress

Amorphous silica nanoparticles (SiNPs) are being utilized in different fields such as medicine, cosmetics, and foods. However, the causes and mechanisms underlying SiNP testicular damage remain largely unclear. In the present study, we aimed to investigate this issue. Thirty male rats were randomly divided into three groups: control group (n = 10), 500 ppm SiNP–treated group (n = 10), and 1000 ppm SiNP–treated group (n = 10). SiNPs were given orally in drinking water for 30 days. Micronucleus assay was performed on blood RBCs. The concentrations of testicular malondialdehyde (MDA) and glutathione (GSH) and catalase (CAT) activity were measured. Moreover, the histopathological alterations and the expression of apoptotic (caspase-3) and pro-inflammatory and oxidative stress markers (iNOS) in testes and epididymis were analyzed and compared between the three groups. The results showed an increased level of micronucleus frequencies in the 1000 ppm–treated group, as well as increased levels of MDA and decreased activity of CAT and GSH content in testicular tissues in the 1000 ppm–treated group, suggesting DNA damage and oxidative stress mechanisms. Also, there were significant testicular histopathological alterations in this group. Furthermore, 1000-ppm SiNPs could enhance testicular apoptosis, inflammation, and oxidative stress by increasing the expression of apoptotic, pro-inflammatory, and oxidative stress genes including caspase 3 and iNOS in the examined tissue. The lower concentration of SiNPs did not produce any significant biochemical, histopathological, or immunohistochemical alterations whereas 1000-ppm SiNPs resulted in significant testicular changes by exacerbating apoptotic, inflammatory, and oxidative stress–mediated testicular damage.

387-P: Novel Mitochondrial Complex I Modulator Reduces Diabetes-Associated Expression of Inflammation and Fibrosis Markers in Human Proximal Tubular Epithelial Cells

Diabetic kidney disease (DKD) is a chronic microvascular complication of diabetes and accounts for 30-40% of all diabetic patients. Patients with DKD experience a progressive and irreversible decline of renal function until dialysis and kidney transplantation become the last line of intervention. Renal loss of function is driven by the diabetic milieu causing an imbalance between the generation of reactive oxygen species (ROS) and the antioxidant defense mechanisms. The excessive amounts of ROS, driven either by mitochondria or by NADPH oxidases (NOXs) , cause inflammation and eventually lead to fibrosis and kidney failure. Methods: To address this problem, we cultivated healthy primary human renal cells (renal proximal tubular epithelial cells) under diabetic conditions. The cells were treated either with 5mM normal glucose (NG) , or 25mM high glucose with 10ng/ml TNF-α or TGF-β (HG) for 24h. In addition, a novel mitochondrial complex I modulator (BIM, 250nM) that has been designed to interfere with the ROS production at mitochondrial complex I has been tested in combination with the treatments. We analyzed the gene expression of different oxidative stress, inflammation as well as fibrosis markers. Results: On the transcriptional level, a treatment with BIM resulted in a significant downregulation of the HG treatment-induced expression of the oxidative stress markers SOD2 (by 40%) , NOX4 (by 61%) and NOX5 (by 34%) , the inflammation markers IL1B (by 29%) and TNFA (by 22%) , as well as the fibrosis markers TGFB1 (by 23%) , COL4A1 (by 50%) and CTGF (by 43%) , as determined by a two-way ANOVA and Tukey’s multiple comparison. Conclusion: The reduction of these markers after treatment with the complex I modulator suggests renoprotective properties in vitro. Further in vivo experiments in diabetic db/db mice will reveal whether the novel mitochondrial complex I modulator can attenuate renal injury in DKD. Disclosure A.Aboolian: None. S.Urner: None. M.Roden: Advisory Panel; Eli Lilly and Company, Research Support; Boehringer Ingelheim International GmbH, Nutricia, Speaker's Bureau; Novo Nordisk. K.Jandeleit-dahm: None.

Expression of Oxidative Stress and Inflammation-Related Genes in Nasal Mucosa and Nasal Polyps from Patients with Chronic Rhinosinusitis.

Chronic rhinosinusitis (CRS) is a prevalent, multifaceted inflammatory condition affecting the nasal cavity and the paranasal sinuses, frequently accompanied by formation of nasal polyps (CRSwNP). This apparently uniform clinical entity is preceded by heterogeneous changes in cellular and molecular patterns, suggesting the presence of multiple CRS endotypes and a diverse etiology. Alterations of the upper airway innate defense mechanisms, including antimicrobial and antioxidant capacity, have been implicated in CRSwNP etiology. The aim of this study was to investigate mRNA expression patterns of antioxidative enzymes, including superoxide dismutase (SOD) and peroxiredoxin-2 (PRDX2), and innate immune system defense players, namely the bactericidal/permeability-increasing fold-containing family A, member 1 (BPIFA1) and PACAP family members, particularly adenylate-cyclase-activating polypeptide receptor 1 (ADCYAP1) in nasal mucosa and nasal polyps from CRSwNP patients. Additional stratification based on age, sex, allergic comorbidity, and disease severity was applied. The results showed that ADCYAP1, BPIFA1, and PRDX2 transcripts are differentially expressed in nasal mucosa and scale with radiologically assessed disease severity in CRSwNP patients. Sinonasal transcriptome is not associated with age, sex, and smoking in CRSwNP. Surgical and postoperative corticosteroid (CS) therapy improves endoscopic appearance of the mucosa, but variably reverses target gene expression patterns in the nasal cavity of CRSwNP patients. Transcriptional cross-correlations analysis revealed an increased level of connectedness among differentially expressed genes under inflammatory conditions and restoration of basic network following CS treatment. Although results of the present study imply a possible engagement of ADCYAP1 and BPIFA1 as biomarkers for CRSwNP, a more profound study taking into account disease severity and CRSwNP endotypes prior to the treatment would provide additional information on their sensitivity.

Do Sleep Disturbances have a Dual Effect on Alzheimer's Disease?

Sleep disturbances and Alzheimer's disease have deleterious effects on various physiological and cognitive functions including synaptic plasticity, oxidative stress, neuroinflammation, and memory. In addition, clock genes expression is significantly altered following sleep disturbances, which may be involved in the pathogenesis of Alzheimer's disease. In this review article, we aimed to discuss the role of sleep disturbances and Alzheimer's disease in the regulation of synaptic plasticity, oxidative stress, neuroinflammation, and clock genes expression. Also, we aimed to find significant relationships between sleep disturbances and Alzheimer's disease in the modulation of these mechanisms. We referred to the controversial effects of sleep disturbances (particularly those related to the duration of sleep deprivation) on the modulation of synaptic function and neuroinflammation. We aimed to know that, do sleep disturbances have a dual effect on the progression of Alzheimer's disease? Although numerous studies have discussed the association between sleep disturbances and Alzheimer's disease, the new point of this study was to focus on the controversial effects of sleep disturbances on different biological functions, and to evaluate the potential dualistic role of sleep disturbances in the pathogenesis of Alzheimer's disease.