SOD2 Gene Expression Research Articles

Targeting high glucose-induced epigenetic modifications at cardiac level: the role of SGLT2 and SGLT2 inhibitors

BackgroundSodium-glucose co-transporters (SGLT) inhibitors (SGLT2i) showed many beneficial effects at the cardiovascular level. Several mechanisms of action have been identified. However, no data on their capability to act via epigenetic mechanisms were reported. Therefore, this study aimed to investigate the ability of SGLT2 inhibitors (SGLT2i) to induce protective effects at the cardiovascular level by acting on DNA methylation.MethodsTo better clarify this issue, the effects of empagliflozin (EMPA) on hyperglycemia-induced epigenetic modifications were evaluated in human ventricular cardiac myoblasts AC16 exposed to hyperglycemia for 7 days. Therefore, the effects of EMPA on DNA methylation of NF-κB, SOD2, and IL-6 genes in AC16 exposed to high glucose were analyzed by pyrosequencing-based methylation analysis. Modifications of gene expression and DNA methylation of NF-κB and SOD2 were confirmed in response to a transient SGLT2 gene silencing in the same cellular model. Moreover, chromatin immunoprecipitation followed by quantitative PCR was performed to evaluate the occupancy of TET2 across the investigated regions of NF-κB and SOD2 promoters.ResultsSeven days of high glucose treatment induced significant demethylation in the promoter regions of NF-kB and SOD2 with a consequent high level in mRNA expression of both genes. The observed DNA demethylation was mediated by increased TET2 expression and binding to the CpGs island in the promoter regions of analyzed genes. Indeed, EMPA prevented the HG-induced demethylation changes by reducing TET2 binding to the investigated promoter region and counteracted the altered gene expression. The transient SGLT2 gene silencing prevented the DNA demethylation observed in promoter regions, thus suggesting a role of SGLT2 as a potential target of the anti-inflammatory and antioxidant effect of EMPA in cardiomyocytes.ConclusionsIn conclusion, our results demonstrated that EMPA, mainly acting on SGLT2, prevented DNA methylation changes induced by high glucose and provided evidence of a new mechanism by which SGLT2i can exert cardio-beneficial effects.Graphical

Phospho-mimetic cardiac troponin I Ser43/45 produces early changes in mitochondria.

Our work tests whether chronic protein kinase C (PKC)-mediated phosphorylation of cardiac troponin I (cTnI) Ser43/45 diminishes cardiac function and initiates progressive heart failure via early increases in oxidative stress and mitochondria. The current work focuses on early gene expression responses in mice after cardiac-specific phospho-mimetic cTnI S43/45D (SD) replacement of endogenous cTnI. Dose-dependent changes in cTnISD replacement do not significantly change function, when measured by echocardiography at 1 month (mos) of age in male or female mice. However, by 2 mos of age, the highest cTnISD replacement line develop significant dysfunction and 100% mortality by 16 weeks of age, with males (M) dying earlier than females (F; F = 14.6 ± 0.4 wks, n = 32; M = 9.3 ± 0.3 wks, n = 68). Based on gene expression measured by quantitative PCR (QPCR) analysis, 1 mos old hearts from this high replacement cTnISD line of mice develop significant reductions in mitochondrially-encoded cytochrome oxidase and cytochrome b in males, which are followed by similar changes at 2 mos of age in females. Reductions in gene expression of critical mitochondrial proteins PGC-1α and SOD2 also develop by 1 mos of age in males. In contrast, there are no detected changes in gene expression of the endoplasmic reticulum (ER) and nuclear stress markers, ATF6 and Nrf2, respectively for males and/or females by 2 mos of age in the high replacement cTnISD mice. Ongoing work is determining whether similar changes develop in the protein expression of these genes in young adult mice. The present results are consistent with the idea that chronic modification of the sarcomere produces a stress signal leading to early mitochondrial and oxidative stress responses, which are predicted to lay the foundation for the progressive deterioration in cardiac function observed in this model.

Identification of a Potential SNP Related to the Expression of Immune Genes and Its Possible Application to Selection of WSSV-Resistant Pacific White Shrimp (<I>Litopenaeus vannamei</I>)

Highlight Research A molecular marker for shrimp selection in disease resistance. Single nucleotide polymorphisms (SNP) in the ALF gene strongly correlate with shrimp resistance to WSSV infection. Higher ALF gene expression in survivor shrimp. SNP as molecular marker inherited in the first generation (G1) shrimp. ARMS-PCR method successfully detect SNP in the shrimp ALF gene. Abstract The Pacific white shrimp (Litopenaeus vannamei) is Indonesia’s main export commodity, but its production is constrained by the white spot syndrome virus (WSSV). Selective breeding of disease-resistant broodstock based on single nucleotide polymorphism (SNP) in the anti-lipopolysaccharide factor (ALF) gene is an alternative strategy for solving the disease problem. This study aimed to detect the SNP g.455 A>G in the anti-lipopolysaccharide factor (ALF) shrimp gene, evaluate the correlation of SNP with WSSV-resistance trait, analyze the expression level of immunity genes and genotype frequencies of the WSSV-resistance population shrimp and analyze the SNP inheritance in the first generation of selected shrimp. A total of 120 individuals from 4 families were used to detect the SNP marker using tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). The correlation of the SNP marker with survival rate (SR) was analyzed using a general linear model (GLM) between genotype frequencies and SR. Genotypic similarities between broodstock and pedigree were analyzed using Chi-square. SNP g.455 A>G was successfully detected using the ARMS-PCR method and had a strong correlation between the marker and SR (p-value of AA = 0.012; AG = 0.359, and GG = 0.001). The resistant population has significantly higher ALF and SOD gene expression levels and AA genotype frequency. The SNP marker was inherited, so the broodstock and pedigree have the same genotype frequencies according to chi-square analysis (χ2 = 0.46 and p-value = 0.497). These results suggested that the g.455 genotype AA could be selected to produce WSSV-resistant Pacific white shrimp.

Biosynthesized magnesium oxide nanoparticles from Tamarindus indica seed attenuate doxorubicin-induced cardiotoxicity by regulating biochemical indexes and linked genes

The phytochemicals of Tamarindus indica seed hydroalcoholic extract were exploited as a biocatalyst for the sustainable synthesis of magnesium oxide nanoparticles (MgO-NPs). This research investigated the cardioprotective effects of biosynthesized magnesium oxide nanoparticle (MgO-NPs). The biosynthesized seed MgO-NPs were characterized by ultraviolet-visible spectroscopy (UV–Vis), X-ray diffraction (XRD), scanning electron microscopy (SEM) with energy-dispersive X-ray diffraction (EDX), and Fourier-transform infrared spectroscopy (FT-IR). These methodological approaches demonstrated their capacity to synthesize crystalline and aggregated MgO-NPs with a size average of 13.38 ± 0.16 nm. The biogenic MgO-NPs were found to have a significant quantity of total phenolic contents (TPC) and total flavonoid contents (TFC), indicating the existence of phenol and flavonoid-like components. The biogenic MgO-NPs demonstrated a significant free radical scavenging effects compared to different standards as measured by the inhibition of free radicals produced in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+), and Nitric oxide (NO) scavenging methods; they also exhibited higher ferric ion reducing capacity in FRAP assay. Moreover, they were found to be non-toxic in cytotoxic assessment. Pretreatment of Wistar Albino rats with seed MgO-NPs resulted in a significant reduction of cardiac biomarkers, i.e., cardiac Troponin-I (cTnI), creatine kinase (CK-MB), and aspartate aminotransferase (AST). The seed MgO-NPs were more successful in reducing lipid levels. The results of the mRNA expression analysis showed that seed MgO-NPs efficiently reduced the expression of the apoptotic genes p53 and Caspase-3 while restoring the expected levels of SOD gene expression. The histopathological observations were primarily focused on the disruption of cardiac fibers and myofibrillar disintegration, which are consistent with the biochemical findings. Therefore, our research suggests that MgO-NPs derived from the seeds of Tamarindus indica as a powerful antioxidant; the administration may be effective in protecting the heart from DOX-induced cardiotoxicity.

In ovo protective effects of chicoric and rosmarinic acids against Thiacloprid-induced cytotoxicity, oxidative stress, and growth retardation on newly hatched chicks

Thiacloprid (TH) is a neonicotinoid insecticide employed in agriculture to protect fruits and vegetables against different insects. It showed different deleterious effects on the general health of non-target organisms including birds and animals, however, its developmental toxicity has yet to be fully elucidated. Chicoric (CA) and rosmarinic (RA) acids are polyphenolic compounds with a wide range of beneficial biological activities. In this study, the possible protective effects of CA and RA were investigated in chick embryos exposed in ovo to TH (1µg/egg) with or without CA (100 µg/egg) or RA (100 µg/egg) co-exposure. TH reduced the hatchling body weight, body weight/egg weight, and relative weight of bursa of Fabricius in the one-day-old hatchlings. Examination of the 7-day-old chicks revealed a decline in feed intake, daily weight gain, feed conversion ratio (FCR), and plasma levels of T3, T4, and growth hormone. Serum ALT, AST activities, and total cholesterol levels showed significant elevations. Hepatic MDA was increased with a reduction in SOD activity and GSH level and downregulation of the liver SOD and GST gene expression pattern. Serum IgG and IgM levels were reduced, and various histopathological alterations were noticed in the liver. Co-administration of CA or RA with TH mitigated the toxic effects on hatchlings. When both CA and RA are combined, they present a synergistic protective effect. CA and RA can be used as protective agents against TH toxicity as they improve growth performance and have hepatoprotective and immunostimulant effects in newly hatched chicks.

Mechanistic role of epigallocatechin-3-gallate in regulation of the antioxidant markers in ethanol induced liver damage in mice

ABSTRACT Background: Failure of endogenous antioxidant system to eliminate high levels of ROS results to oxidative stress, consequently leading to liver damage. Excessive alcohol consumption leads to liver damage due to ROS generation, thus regarded as the leading causes of death worldwide, suggesting great need for use of exogenous antioxidants like epigallocatechin-3-gallate (EGCG), to avert liver damage. However, underlying EGCG mechanisms remains elusive.Aim of the work: This study focused on mechanistic role of EGCG in regulation of antioxidant marker activities during ethanol induced liver damage.Materials and Methods: 20 male Swiss Albino mice were divided into four groups and treated with different dosage. ALT, AST and total proteins were determined as indicators of liver damage. MDA and protein carbonyls were measured as oxidative stress markers. Activities of SOD and CAT were determined. SOD-2 and CAT differential gene expression were also determined. Liver histology analysis by H&E staining. All the experiments were run in duplicates. ANOVA was used to analyzes data using Tukey’s multiple comparison tests and results considered statistically significant if p ≤ 0.05 at 95% confidence level. Results: Findings revealed that chronic consumption of ethanol leads to liver damage through increased levels of serum ALT and AST, MDA, protein carbonyls and remarkable diffuse lipid droplets and decreased enzyme activity of SOD and CAT. EGCG increased activity of SOD and CAT and SOD-2 expression and did not affect CAT expression. Conclusion: In summary, ethanol induces liver damage, and administration of EGCG increases antioxidant system expression, suggesting its role in regulating their activities, defensive of oxidative stress through various pathways.

Oxidized Dietary Oil, High in Omega-3 and Omega-6 Polyunsaturated Fatty Acids, Induces Antioxidant Responses in a Human Intestinal HT29 Cell Line.

When oxidized, dietary oils generate products which have the potential to cause adverse effects on human health. The objective of the study was to investigate whether lipid oxidation products in an oxidized dietary oil can be taken up in intestinal cells, induce antioxidant stress responses and potentially be harmful. The in vitro cell model HT29 was exposed to camelina oil with different extents of oxidation, or only 4-hydroxy-2-hexenal (HHE) or 4-hydroxy-2-nonenal (HNE). The cellular content of HHE increased with an increasing extent of oxidation of the camelina oil added to the cell's growth media, whereas HNE did not show a similar trend. Deuterated HHE was taken up by the HT29 cells, with 140 µM HHE metabolized within 0.5-1 h. The low oxidation degree of the camelina oil increased the gene expression of antioxidant markers (GPX, ATF6, XBP1). The increase in the gene expression of SOD at medium oxidation levels of the oil might indicate different regulation mechanisms. Highly oxidized camelina oil and a low concentration of HHE, over time, induced SOD and catalase enzyme activity in HT29 cells. Oxidized camelina oil contains multiple oxidation products which can be responsible for the intracellular responses observed in HT29 cells, while HHE and HNE in combination with other oxidation products induce antioxidant defence responses.

Glycerol Monolaurate Alleviates Oxidative Stress and Intestinal Flora Imbalance Caused by Salinity Changes for Juvenile Grouper.

Groupers with an initial body weight of 9.10 ± 0.03 g were selected to investigate whether dietary addition of 0 (G0) and 1800 mg/kg glycerol monolaurate (GML, G1800) could alleviate the oxidative stress response and intestinal flora imbalance after 0, 6, 12, and 24 h of salinity change in grouper. Experimental results show that the dietary addition of GML significantly reduced the liver MDA content and increased the SOD activity of grouper. The gene expression of CAT and SOD increased and then decreased with time after adding 1800 mg/kg GML, and the highest values were significantly higher than those of the control group. Salinity change had a slight effect on the top four intestinal flora composition of grouper at 0, 12, and 24 h, with changes occurring only at 6 h when Cyanobacteria replaced Actinobacteria. The addition of dietary GML slowed down the intestinal flora disorder, inhibited the colonization of harmful bacterium Vibrio, and promoted the abundance of beneficial bacterium Bacillus. In conclusion, dietary GML significantly reduced the oxidative damage caused by sudden changes in salinity, improved the antioxidant capacity, and alleviated the intestinal flora imbalance in juvenile grouper.

Overexpression of Iron Super Oxide Dismutases A/B Genes Are Associated with Antimony Resistance of Leishmania tropica Clinical Isolates.

Pentavalent antimonial has been a drug of choice against leishmaniasis, despite the emergence of treatment failure. Identification of resistance markers is urgently needed to design new therapeutic strategies. Iron-Superoxide dismutases (Fe-SODs) are antioxidant enzymes contributing to detoxify reactive oxygen species to prevent a cell from oxidative stress. Since antimonial compounds induce oxidative stress, in this survey, the expression of SOD genes was investigated to identify their expression pattern in clinical resistant isolates. This cross-sectional survey was done in Mashhad City, northeast of Iran during 2014 to 2019. The RNA expression level of mitochondrial (SODA) and glycosomal (SODB) superoxide dismutase was investigated in 25 antimony responsive (n=15) and unresponsive (n=10) anthroponotic cutaneous leishmaniasis (ACL) patients. Total RNA extraction and cDNA synthesis, the qRT-PCR approach was utilized to investigate the relative RNA expression level. The transcript level of SODs was over-expressed in the most resistant isolates. Gene expression analysis demonstrated the over-expression of SODA and B by a factor of 3.8 and 4.81, respectively, in resistance isolates vs. sensitive ones. Aberrant expression of SODA/B in unresponsive parasites could potentially implicate in detoxifying antimony-induced oxidative stress. Moreover, SODs might be considered as potential predictive markers of the response to antimonials in ACL patients in endemic areas.

Alterations in the Expression of Antioxidant Enzyme Genes in Response to Pesticide Exposure During Embryonic Development in the Native Reptile Species Caiman latirostris

The aim of this study was to quantify the expression levels of Catalase (cat) and copper, zinc Superoxide dismutase (Cu, Zn-sod) genes involved in the antioxidant response in Caiman latirostris (broad-snouted caiman) blood, after embryonic exposure to the formulations cypermethrin (CYP), chlorpyrifos (CPF), glyphosate (GLY), and their binary and ternary mixtures. Experimental groups were: negative control (NC-distilled water), vehicle control (VC-ethanol), GLY-2%, CYP- 0.12%, CPF- 0.8%, a ternary mixture of them (TM), and three binary mixtures. The applications were made on the nest material in contact with the eggs at the beginning of the incubation period. After hatching, RNA was isolated from blood and expression levels analyzed through qPCR. The results showed downregulation in the expression of sod and cat genes in the three binary mixtures studied, compared to the controls. In addition, we found a possible antagonistic effect between different pesticides in the TM on the expression of both genes.

MLN-4760 Induces Oxidative Stress without Blood Pressure and Behavioural Alterations in SHRs: Roles of Nfe2l2 Gene, Nitric Oxide and Hydrogen Sulfide.

Reduced angiotensin 1-7 bioavailability due to inhibition of angiotensin-converting enzyme 2 (ACE2) may contribute to increased mortality in hypertensive individuals during COVID-19. However, effects of ACE2 inhibitor MLN-4760 in brain functions remain unknown. We investigated the selected behavioural and hemodynamic parameters in spontaneously hypertensive rats (SHRs) after a 2-week s.c. infusion of MLN-4760 (dose 1 mg/kg/day). The biochemical and molecular effects of MLN-4760 were investigated in the brainstem and blood plasma. MLN-4760 had no effects on hemodynamic and behavioural parameters. However, MLN-4760 increased plasma hydrogen sulfide (H2S) level and total nitric oxide (NO) synthase activity and conjugated dienes in the brainstem. Increased NO synthase activity correlated positively with gene expression of Nos3 while plasma H2S levels correlated positively with gene expressions of H2S-producing enzymes Mpst, Cth and Cbs. MLN-4760 administration increased gene expression of Ace2, Sod1, Sod2, Gpx4 and Hmox1, which positively correlated with expression of Nfe2l2 gene encoding the redox-sensitive transcription factor NRF2. Collectively, MLN-4760 did not exacerbate pre-existing hypertension and behavioural hyperactivity/anxiety in SHRs. However, MLN-4760-induced oxidative damage in brainstem was associated with activation of NO- and H2S-mediated compensatory mechanisms and with increased gene expression of antioxidant, NO- and H2S-producing enzymes that all correlated positively with elevated Nfe2l2 expression.

Effect of nano-particles of zinc oxide and selenium on antioxidant status, aminotransferase enzymes activities and genes expression of sod-1 and vg in honey bee during the hot season

BackgroundThe nano-particle of zinc oxide (nZn) and selenium (nSe) has recently attracted in animal feeding because of their high absorbability, catalytic efficiency, and low toxicity than organic and non-organic forms. The present study was done to evaluate the effect of nZn and nSe supplementation in the honey bee's diet under heat stress on antioxidant status, aminotransferase activity, and the relative gene expression of vg and sod-1. Materials and methodsFifty honey bee colonies were randomly divided into five treatments and 10 replicates. Treatments were sugar syrup without or with nanoparticle supplementation, as: i) the control group (no additives); ii) 100 µgL−1 nano-zinc oxide (nZn-1); iii) 200 µg L − 1 of nano-zinc oxide (nZn-2); iv) 100 µg L − 1 nano-selenium (nSe-1); v) 200 µg L − 1 nano-selenium (nSe-2). On days 30 and 60 of experiment, the bee's samples were taken and antioxidant status, aminotransferase activities and relative genes expression of vg and sod-1 were measured. ResultsThe highest (P<0.05) body weight and the lowest (P<0.05) malondialdehyde level were observed in bee colonies receiving nZn-2 and nSe-1. Body weight of newly emerged bee in the control group, nZn-1 and nSe-1 was 110, 122, and 119 mg, respectively. Total antioxidant capacity increased (P<0.05) as the dose of nZn increased, but it decreased (P<0.05), as the dose of nSe increased. The relative genes expression of vg and sod-1 increased (P<0.05) as the dose of nZn increased. nSe increased gene expression of sod-1, but it decreased (P<0.05) gene expression of vg as dose of nSe increased. Aminotransferase enzymes activities increased (P<0.05), as the dose of nZn increased, but their activities in the bees receiving nSe decreased as the dose of nSe increased. ConclusionThe inclusion of nZn at dose of 200 µg L − 1 in syrup fed to honey bee is recommended to increase the antioxidant status, and body weight of newly emerged bees. Supplementation of nSe at dose of 50 µg L − 1 has beneficial effects on the antioxidant status and body weight of newly emerged bees, and doses higher than 50 µg L − 1 is not recommended.

Chiral β‐arylchalcogenium azide induce apoptosis and regulate Oxidative Damage on Human Bladder Cancer Cells

AbstractIn the present study, we aimed to evaluate the cytotoxic effect of chiral β‐Aryl‐chalcogenium azide compounds in vitro against the human bladder cancer cell line 5637. The cell growth inhibition was evaluated by MTT assay. The apoptosis induction and cellular viability were measured by DAPI staining and live/dead assay, respectively. Also, gene expression of pro‐apoptotic, anti‐apoptotic and oxidative stress pathways were investigated by qRT‐PCR. Finally, in silico molecular docking between the compounds and proteins involved in apoptotic and cell growth pathways was performed as well. Our results showed that 5c [(S)‐(2‐azido‐1‐phenyl‐3‐telurophenyl)‐propane)] and 5j [(S)‐(2‐azido‐1‐phenyl‐3‐teluro‐p‐methoxy‐phenyl)‐propane)] compounds presented concentration‐dependent cytotoxicity. Apoptosis analysis showed chromatin condensation in 13 % and 20.3 % of cells treated with compounds 5 c and 5 j, respectively, compared to untreated cells (2.34 %). Moreover, 5 c increased the relative expression of p53, Caspase‐3, Caspase‐9, SOD and CAT genes. Meanwhile, 5 j increased the expression of p53, p21, Caspase‐3, Caspase‐9, Bax, SOD, CAT, GPx, GR and iNOS genes. Finally, molecular docking showed that 5 c has a binding affinity to Survivin, Bcl‐XL, RSK2, SGK1, while 5 j showed binding affinity to the SGK1 protein only. These findings suggest that β‐Aryl‐chalcogenium azide compounds could have a potential antitumoral effect in bladder cancer cells in vitro.

Combined Effects of Temperature and Salinity Affect the Surviv-Al of Asian Green Mussel (Perna viridis) through Digestive and Antioxidant Performance.

Changes in temperature and salinity of the marine environment fluctuate continuously, and the effects of these changes on shellfish survival are significant. In this study, the survival rate of adult Asian green mussels (Perna viridis) was measured after short-term treatments (7 days) for a range of temperature (20 °C, 25 °C, 30 °C) and salinity (23‰, 28‰, 33‰). The digestive (amylase, lipase, trypsin and pepsin activities) and antioxidant performance (SOD and GPX gene expression; SOD, GPX and CAT activities; GSH and MDA Concentration) were measured and MANOVA results were obtained using a generalized linear model between certain factors (temperature (T), salinity (S) and temperature × salinity (T × S)) and survival rate. Both T and T × S significantly affected the survival rate of Asian green mussels. The overall relationship between the survival rate of P. viridis and T and S within the experimental range can be summarised by the equation: ln(Survival rate) = 54.9282 − 2.5627 × T − 3.6180 × S + 0.1857 × T × S + 0.0156 × T2 + 0.0520 × S2 − 0.0012 × T2 × S − 0.0023 × T × S2. The optimal temperature–salinity combination was 23.698 °C/30.760‰. T, S and T × S all had a significant impact on amylase and lipase activities of Asian green mussels. In the hepatopancreas, gill, mantle, adductor muscle, gonad and foot tissues of Asian green mussel, the SOD and GPX gene expression were responsive to temperature–salinity changes. The antioxidant performance of the combinations far from the optimal temperature–salinity combination were significantly improved. Our results suggest that combined temperature and salinity effects have a regular impact on the survival of Asian green mussels and that there is a link between survival and digestive and antioxidant performance.

Morinda officinalis Polysaccharides Ameliorates Bone Growth by Attenuating Oxidative Stress and Regulating the Gut Microbiota in Thiram-Induced Tibial Dyschondroplasia Chickens.

Tibial dyschondroplasia (TD) occurs in chickens and other fast-growing birds, affecting their cartilage growth and leading to reduced meat quality in broilers. Morinda officinalis polysaccharide (MOP) is one of the chief active components of Morinda officinalis, which promotes bone formation, inhibiting bone loss and having anti-oxidant and anti-inflammatory properties. A total of 120 AA chickens were randomly divided into the CON group (basal diet), TD group (100 mg/kg thiram + basal diet), and MOP group (100 mg/kg thiram + basal diet + water with 500 mg/kg MOP). The experiment lasted 21 days. The results showed that MOP could alleviates broiler lameness caused by TD, restore the morphological structure of tibial growth plate (TGP), increase tibial weight (p < 0.05), balance the disorder of calcium and phosphorus metabolism, and promote bone formation by increasing the expression of BMP-2, Smad4, and Runx2 genes In addition, MOP supplementation stimulated the secretion of plasma antioxidant enzymes (T-SOD and GSH-Px) by regulating the expression of SOD and GPX-1 genes, thereby enhancing the antioxidant capacity of TD broilers. Interestingly, we observed MOP can also improve gut microbiota by increasing the beneficial bacteria count and decreasing the harmful bacteria count. These findings indicated that MOP can regulate bone formation through the BMP/Smads signaling pathway, attenuating oxidative stress and regulating the gut microbiota of TD broilers, so as to achieve the effect of treating TD. This suggests that MOP might be a potential novel drug in the treatment of TD in chickens.

Maternal hypothyroidism causes oxidative stress and endoplasmic reticulum stress in the maternal-fetal interface of rats.

Maternal hypothyroidism is associated with pre-eclampsia and intrauterine growth restriction, gestational diseases involving oxidative stress (OS) and endoplasmic reticulum stress (ERS) in the placenta. However, it is not known whether hypothyroidism also causes OS and ERS at the maternal-fetal interface. The aim was to evaluate the fetal-placental development and the expression of mediators of OS and of the unfolded protein response (UPR) in the maternal-fetal interface of hypothyroid rats. Hypothyroidism was induced in Wistar rats with propylthiouracil and the fetal-placental development and placental and decidual expression of antioxidant, hypoxia, and UPR mediators were analyzed at 14 and 18 days of gestation (DG), as well the expression of 8-OHdG and MDA, and reactive oxygen species (ROS) and peroxynitrite levels. Hypothyroidism reduced fetal weight at 14 and 18 DG, in addition to increasing the percentage of fetal death and reducing the weight of the uteroplacental unit at 18 DG. At 14 DG, there was greater decidual and/or placental immunostaining of Hif1α, 8-OHdG, MDA, SOD1, GPx1/2, Grp78 and CHOP in hypothyroid rats, while there was a reduction in placental and/or decidual gene expression of Sod1, Gpx1, Atf6, Perk, Ho1, Xbp1, Grp78 and Chop in the same gestational period. At 18 DG, hypothyroidism increased the placental ROS levels and the decidual and/or placental immunostaining of HIF1α, 8-OHdG, MDA, ATF4, GRP78 and CHOP, while it reduced the immunostaining and enzymatic activity of SOD1, CAT, GST. Hypothyroidism increased the placental mRNA expression of Hifα, Nrf2, Sod2, Gpx1, Cat, Perk, Atf6 and Chop at 18 DG, while decreasing the decidual expression of Sod2, Cat and Atf6. These findings demonstrated that fetal-placental restriction in female rats with hypothyroidism is associated with hypoxia and dysregulation in placental and decidual expression of UPR mediators and antioxidant enzymes, and activation of oxidative stress and endoplasmic reticulum stress at the maternal-fetal interface.

Protective Effects of PollenAid Plus Soft Gel Capsules' Hydroalcoholic Extract in Isolated Prostates and Ovaries Exposed to Lipopolysaccharide.

Pollen extract represents an innovative approach for the management of the clinical symptoms related to prostatitis and pelvic inflammatory disease (PID). In this context, the aims of the present work were to analyze the phenolic composition of a hydroalcoholic extract of PollenAid Plus soft gel capsules, and to evaluate the extract’s cytotoxic effects, in human prostate cancer PC3 cells and human ovary cancer OVCAR-3 cells. Additionally, protective effects were investigated in isolated prostate and ovary specimens exposed to lipopolysaccharide (LPS). The phytochemical investigation identified catechin, chlorogenic acid, gentisic acid, and 3-hydroxytyrosol as the prominent phenolics. The extract did not exert a relevant cytotoxic effect on PC3 and OVCAR-3 cells. However, the extract showed a dose-dependent inhibition of pro-inflammatory IL-6 and TNF-α gene expression in prostate and ovary specimens, and the extract was effective in preventing the LPS-induced upregulation of CAT and SOD gene expression, which are deeply involved in tissue antioxidant defense systems. Finally, a docking approach suggested the capability of catechin and chlorogenic acid to interact with the TRPV1 receptor, playing a master role in prostate inflammation. Overall, the present findings demonstrated anti-inflammatory and antioxidant effects of this formulation; thus, suggesting its capability in the management of the clinical symptoms related to prostatitis and PID.

Simvastatin and fluvastatin attenuate trauma-induced cell death and catabolism in human cartilage.

Joint injuries are known to induce pathomechanisms that might lead to posttraumatic osteoarthritis (PTOA). In this regard, statins with their pleiotropic effects could represent potential therapeutic agents in preventing the development of PTOA. Therefore, we investigated the effects of simvastatin and fluvastatin in a drop-tower-based human ex vivo cartilage trauma model. After 7 days, a mechanical impact (0.59 J) resulted in a decrease of the cell viability and increased expression of catabolic enzymes in cartilage explants. Simvastatin and fluvastatin treatment of impacted cartilage demonstrated cell protective effects in a concentration dependent manner. Moreover, statin therapy exhibited chondroprotective effects as demonstrated by attenuated expression of MMP-2 and MMP-13 as well as subsequent breakdown of collagen type II (after impact). Further analysis indicated antioxidative properties of the statins by upregulating the gene expression of SOD2 and suppression that of NOX2 and NOX4. Despite its protective effects, simvastatin impaired the biosynthesis of collagen type II, which was confirmed during chondrogenic redifferentiation of high passage chondrocytes. However, while long-term administration of statins for 4 weeks impaired chondrogenic redifferentiation, addition of simvastatin at low concentrations for 1 week exhibited a slightly promoting effect. In conclusion, our data imply that simvastatin and fluvastatin are suitable in terms of initial harm reduction after cartilage trauma.

Testicular Expression of Antioxidant Enzymes and Changes in Response to a Slow-Release Deslorelin Implant (Suprelorin® 4.7 mg) in the Dog.

Simple SummaryWe investigated the expression of the testicular antioxidant enzyme system, i.e., superoxide dismutase (SOD1 and 2), catalase (CAT), glutathione peroxidase (GPx1), and glutathione disulfide reductase (GSR), in healthy adult and prepubertal dogs and after slow-release GnRH superagonist deslorelin (Suprelorin® 4.7 mg subcutaneous implant, Virbac, France) treatment in adult dogs. qPCR and immunohistochemistry were used to detect gene expression and protein expression and localization, respectively. These antioxidant enzymes were variably expressed within cellular compartments in the seminiferous epithelium and the interstitium. SOD1, CAT, and GSR protein expression were also dependent on the stage of the seminiferous epithelium cycle in normal adult dogs. Deslorelin treatment and prepubertal status affected the expression and localization of the investigated antioxidant enzymes. Our results confirmed the presence of a functional antioxidant enzyme system in the canine testis, which was also observed during deslorelin-induced spermatogenic and steroidogenic arrest, as well as in prepubertal dogs. These results improve our current understanding of the local antioxidant protective mechanisms within the testis in the dog. Spermatogenesis takes place in a hypoxic environment, and antioxidant enzymes protect germ and somatic cells from free radical-mediated damage. Expression of the antioxidant enzyme system in the canine testis has not yet been investigated. We hypothesized that the slow-release GnRH superagonist deslorelin 4.7 mg implant, which induces temporary reversible suppression of endocrine and germinative testicular function, would affect the testicular expression of antioxidant enzymes compared to untreated adult and prepubertal dogs. The goal of this study was to investigate and compare gene (by qPCR, in whole-tissue homogenates) and protein expression (by immunohistochemistry) of superoxide dismutase (SOD1, SOD2), catalase (CAT), glutathione peroxidase (GPx1), and glutathione disulfide reductase (GSR) in the testes of untreated adult (CON, n = 7), prepubertal (PRE, n = 8), and deslorelin-treated (DES, n = 5, 16 weeks after implantation) dogs. We found that in DES dogs, the gene expression of SOD1 was significantly (p < 0.05) lower and GPx1 was higher than in CON, and SOD2 was higher than in PRE. Expression of all, except for the SOD2 mRNA, differed between the CON and PRE dogs. Immunohistochemistry showed distinct cell-specific localization and expression patterns for the antioxidant enzymes in each experimental group. Additionally, in the CON animals, cell-specific SOD1, CAT, and GSR expression was dependent on the stage of the seminiferous epithelium cycle. These findings confirm that members of the antioxidant enzyme system are present in normal adult and prepubertal testis as well as in the deslorelin-treated downregulated adult canine testis, and that this local antioxidant system protects developing germ cells and somatic cells from oxidative damage. Different expression patterns of antioxidant enzymes in various germ cell populations and stages of the seminiferous epithelium cycle may indicate differences in their susceptibility to oxidative stress depending on their developmental and maturation stage. The continued presence of the antioxidant enzymes in the testis of DES dogs offers protection to spermatogonia as well as Sertoli and Leydig cells from oxidative stress during temporary infertility, potentially contributing to ensure the reversibility of suppression and the return of normal spermatogenesis and steroidogenesis after the end of deslorelin treatment.

Kanamycin and Cisplatin Ototoxicity: Differences in Patterns of Oxidative Stress, Antioxidant Enzyme Expression and Hair Cell Loss in the Cochlea.

Kanamycin and cisplatin are ototoxic drugs. The mechanisms are incompletely known. With subcutaneous kanamycin (400 mg/kg, 15 days), auditory threshold shifts were detected at days 12–13 at 16 and 32 kHz, extending to 8 and 4 kHz at days 14–15. The outer hair cell (OHC) loss was concentrated past day 12. The maximum cochlear length showing apoptotic cells, tested with TUNEL, was at day 13. At day 15, 1/5 of the apical cochlea contained preserved OHCs. 3-nitrotyrosine (3-NT) immunolabeling, showing oxidative stress, was found in surviving OHCs and in basal and middle portions of the stria vascularis (SV). The antioxidant Gpx1 gene expression was decreased. The immunocytochemistry showed diminished Gpx1 in OHCs. With intraperitoneal cisplatin (16 mg/kg, single injection), no evoked auditory activity was recorded at the end of treatment, at 72 h. The basal third of the cochlea lacked OHCs. Apoptosis occupied the adjacent 1/3, and the apical third contained preserved OHCs. 3-NT immunolabeling was extensive in OHCs and the SV. Gpx1 and Sod1 gene expression was downregulated. Gpx1 immunostaining diminished in middle and basal SV. More OHCs survived cisplatin than kanamycin towards the apex, despite undetectable evoked activity. Differential regulation of antioxidant enzyme levels suggests differences in the antioxidant response for both drugs.