Superoxide Dismutase mRNA Expression Research Articles

Estrogen Replacement Therapy Induces Antioxidant and Longevity-Related Genes in Women after Medically Induced Menopause.

Females live longer than males in many species, including humans, and estrogens are in part responsible for this protection against aging. We reported previously that estrogens can protect rats against oxidative stress, by inducing antioxidant and longevity-related genes. Thus, this study was aimed at confirming the ability of estrogens to upregulate antioxidant and longevity-related genes in humans. For this purpose, we selected 16 women of reproductive age (18-42 years old) undergoing a fertility treatment that includes a medically induced menopause, at the Valencian Infertility Institute. We took blood samples at each time point of the treatment (basal, induced menopause, estrogen, and estrogen plus progesterone replacement therapy). mRNA expression of antioxidant and longevity-related genes in peripheral blood mononuclear cells (PBMC) was determined by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Determination of reduced glutathione (GSH) in total blood was carried out using high-performance liquid chromatography (HPLC). As expected, we found that medically induced menopause significantly decreased sexual hormone (estrogens and progesterone) levels. It also lowered glutathione peroxidase (GPx), 16S rRNA, P21, and TERF2 mRNA expression and blood GSH levels. Estrogen replacement therapy significantly restored estrogen levels and induced mRNA expression of manganese superoxide dismutase (MnSOD), GPx, 16S rRNA, P53, P21, and TERF2 and restored blood GSH levels. Progesterone replacement therapy induced a significant increase in MnSOD, P53, sestrin 2 (SENS2), and TERF2 mRNA expression when compared to basal conditions. These findings provide evidence for estrogen beneficial effects in upregulating antioxidant and longevity-related genes in women.

Protective Effect of Epigallocatechin-3-Gallate in Hydrogen Peroxide-Induced Oxidative Damage in Chicken Lymphocytes.

Epigallocatechin-3-gallate (EGCG) is one of the fundamental compounds in green tea. The present study was to evaluate the protective effect of EGCG in oxidative damage and apoptosis induced by hydrogen peroxide (H2O2) in chicken lymphocytes. Results showed that preincubation of lymphocytes with EGCG significantly decreased H2O2-reduced cell viability and apoptotic cells with DNA damage, restored the H2O2-dependent reduction in total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), glutathione (GSH), and glutathione disulfide (GSSG), and suppressed the increase in intracellular reactive oxygen species (ROS), nitric oxide (NO), nitric oxide synthesis (NOS), malondialdehyde (MDA), lipid peroxide (LPO), and protein carbonyl (Carbonyl). In addition, preincubation of the cells with EGCG increased mitochondrial membrane potential (MMP) and reduced calcium ion ([Ca2+]i) load. The protective effect of EGCG in oxidative damage in lymphocytes was accompanied by mRNA expression of SOD, Heme oxygenase-1 (HO-1), Catalase (CAT), GSH-PX, nuclear factor erythroid 2-related factor 2 (Nrf2), and thioredoxin-1 (Trx-1). As EGCG had been removed before lymphocytes were challenged with H2O2, the activation of genes such as Nrf2 and Trx-1 by preincubation with EGCG could be the main reason for EGCG to protect the cells from oxidative damage by H2O2. Since oxidative stress is an important mechanism of biological damage and is regarded as the reasons of several pathologies, the present findings may be helpful for the use of tea products to prevent oxidative stress and maintain healthy in both humans and animals.

Glutamine pretreatment protects bovine mammary epithelial cells from inflammation and oxidative stress induced by γ-d-glutamyl-meso-diaminopimelic acid (iE-DAP)

Glutamine (GLN) has many types of biological activity in rats, including anti-inflammatory, antioxidative stress, and anti-apoptosis effects. However, little is known about the effects of GLN on bovine mammary epithelial cells (BMEC). γ-d-Glutamyl-meso-diaminopimelic acid (iE-DAP) is a cell wall peptidoglycan component of gram-negative bacteria that can be recognized by the intracellular receptor nucleotide-binding oligomerization domain-containing protein 1 (NOD1) and can cause bovine mastitis. The goal of the present study was to investigate whether GLN protects BMEC from iE-DAP-induced inflammation, oxidative stress, and apoptosis. We cultured BMEC in a GLN-free medium for 24 h and then separated them into 4 groups: cells treated with 1× PBS for 26 or 32 h (control); cells stimulated by 10 μg/mL iE-DAP for 2 or 8 h (2- or 8-h iE-DAP); cells pretreated with 8 or 4 mM GLN for 24 h followed by 2 or 8 h of 1× PBS treatment (8 or 4 mM GLN); and cells pretreated with 8 or 4 mM GLN for 24 h followed by 2 or 8 h of iE-DAP treatment (DG). In the 2-h iE-DAP group, when levels of inflammation peaked, iE-DAP treatment increased both the mRNA and protein expression of NOD1, inhibitor of nuclear factor-κB (NFKBIA, IκB), and nuclear factor-κB subunit p65 (RELA, NF-κB p65), as well as the mRNA expression of IL6 and IL8 and levels of IL-6 and tumor necrosis factor-α in cell culture supernatants. In contrast, 8 mM GLN pretreatment inhibited the mRNA and protein expression of inflammatory-related factors by suppressing the NOD1/NF-κB pathway. In the 8-h iE-DAP group, iE-DAP treatment decreased the mRNA and protein expression of extracellular regulated kinase (Erk, ERK) and nuclear factor erythroid 2-associated factor2 (NFE2L2, Nrf2), as well as the mRNA expression of superoxide dismutase 1 (SOD1), catalase (CAT), coenzyme II oxidoreductase 1 (NQO1), and heme oxygenase 1 (HMOX1, HO1). In addition, iE-DAP treatment increased the expression of malondialdehyde in BMEC when oxidative stress levels peaked. Interestingly, 4 mM GLN pretreatment induced the mRNA and protein expression of antioxidative stress-related factors and inhibited the expression of reactive oxygen species in BMEC by promoting the ERK/Nrf2 pathway. Moreover, GLN reduced apoptosis caused by inflammation and oxidative stress in BMEC. This is the first report showing that GLN protects against iE-DAP-induced inflammation and oxidative stress via the NOD1/NF-κB and ERK/Nrf2 pathways in BMEC.

Supplementation of cryoprotectant with Pinus massoniana bark extract improves human sperm vitality and fertility potential

AbstractBackgroundSperm cryopreservation is commonly used to preserve fertility but often decreases sperm quality by inducing oxidative stress. Pinus massoniana bark extract (Pinus massoniana, PMBE) exhibits strong antioxidant activity and has been used in traditional Chinese medicine.ObjectivesThe objective was to determine whether adding PMBE to cryoprotectant could improve human sperm quality after cryo‐resuscitation and to investigate the potential regulatory mechanisms.MethodsPMBE was used as a cryoprotectant supplement. Hydrogen peroxide was applied to generate oxidative stress. The changes in sperm quality upon cryo‐resuscitation and hydrogen peroxide treatment were measured by computer‐aided sperm analysis and fluorescein isothiocyanate‐labeled peanut (Pisum sativum) agglutinin staining. Oxidative stress was evaluated by dichlorofluorescein diacetate staining and the malondialdehyde content. The antioxidant capacity was determined by the glutathione peroxidase activity and the reduced glutathione content. Mass spectrometry was utilized to screen factors potentially regulated by PMBE, and the screened factors were evaluated by Western blot. Mouse in vitro fertilization was used to assess the effectiveness and safety of PMBE on the fertilization outcome of damaged spermatozoa.ResultsPMBE addition to cryoprotectant could effectively promote post‐thaw vitality and acrosome reaction rates, reduce intracellular reactive oxygen species, and preserve antioxidant capacity and mitochondrial function in the presence of cryo‐resuscitation and hydrogen peroxide treatment. Decreased in the expression of critical proteins such as inositol‐3‐phosphate synthase 1 (ISYNA1) and AKT1 upon resuscitation was rescued by PMBE supplementation, further reducing the cytoplasmic Nrf2 protein level and markedly increasing the nuclear level. PMBE dramatically enhanced the mRNA and protein expression of superoxide dismutase (SOD2). PMBE effectively alleviated the hydrogen peroxide‐induced decline in the two‐cell embryo rates.ConclusionSupplementing cryoprotectant with PMBE improves human sperm quality after cryo‐resuscitation by reducing oxidative stress and stabilizing the functional proteins ISYNA1 and AKT1. PMBE addition improves the fertilization outcome of oxidative stress‐damaged spermatozoa.

Ulva prolifera Extract Alleviates Intestinal Oxidative Stress via Nrf2 Signaling in Weaned Piglets Challenged With Hydrogen Peroxide.

Background Ulva prolifera extract contains a variety of functional active substances. Whether these substances had any beneficial effects on the small intestine of weaned piglets under oxidative stress remained unknown.MethodWe explored the effects of U. prolifera extract on oxidative stress and related mechanisms in weaned piglets and intestinal porcine epithelial cells (IPEC-J2) challenged with hydrogen peroxide.Results U. prolifera extract was found to mainly consist of polyphenols and unsaturated fatty acids. U. prolifera extract increased total antioxidant capacity and superoxide dismutase (SOD) activity, while it decreased malondialdehyde content, in the serum of weaned piglets challenged with hydrogen peroxide. Moreover, U. prolifera extract increased mRNA expression of SOD and catalase, as well as the intestinal expression of nuclear NF-E2-related factor 2 (Nrf2), both in vitro and in vivo. Furthermore, U. prolifera extract decreased reactive oxygen species and improved mitochondrial respiration in IPEC-J2 cells treated with hydrogen peroxide. However, AMPK inhibition did not affect nuclear Nrf2 expression and only partially affected the effects of U. prolifera extract on oxidative stress.ConclusionWe suggest that U. prolifera extract alleviates oxidative stress via Nrf2 signaling, but independent of AMPK pathway in weaned piglets challenged with hydrogen peroxide. These results shed new insight into the potential applications of U. prolifera extract as a therapeutic agent for the prevention and treatment of oxidative stress-induced intestinal diseases.

The effect of purple sweet potato (Ipomea batatas L) ethanol extract on Estrogen Receptor Alpha (ERα) and SOD mRNA expression in the menopause-liver animal model

Background: Aging and estrogen deficiency in menopause women increase susceptibility to liver diseases. In postmenopausal women, the change of ERα expressions and oxidative stress is related to liver diseases' pathogenesis. Phytoestrogens have been shown to protect and repair the liver organ due to estrogen deficiency. Purple Sweet Potatoes contain anthocyanins, a subgroup of flavonoids that have been shown to have antioxidant and phytoestrogen activity. This study aims to evaluate the phytoestrogen and antioxidant activities of purple sweet potato in a liver animal model of menopause through Superoxide Dismutase (SOD) and ERα expressions.Method: A true experimental study by randomized post-test-only control group design was conducted among 28 female Wistar rats-aged 10-12 weeks with a bodyweight of 180-220 gram, which underwent bilateral ovariectomies. These animals were divided into 4 groups, with 7 rats in each group. On day 7, Group T0, as control was given aquadest, ethanol extract of purple sweet potato was assigned to Group T1, T2, and T3 with doses 1, 2, and 4 mL/day, respectively, during 30 days. The SOD and ERα mRNA expressions were evaluated by qRT-PCR with specific primers. Data were analyzed using SPSS version 20 for Windows.Results: The mRNA expressions of ERα were not significantly different among groups (p>0.05), while SOD expressions were significantly different among groups (p<0.05). The expression of SOD in Group T1 (504.39±231.4 pg/μL), T2 (2,147.92±417.1 pg/μL), and T3 (405.30±224.1 pg/μL) showed a significantly higher than Group T0 (15.34±15.27) (p<0.05)Conclusion: Oxidative stress could be prevented by purple sweet potato in the liver through the SOD mRNA expression but not ERα.

HEREDITARY NEUROPATHIES & ALS: P.110 Development and characterization of in vitro models to test the efficiency of gene therapy approaches in SOD1-linked Amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of motor neurons (MNs) that causes skeletal muscle (SM) paralysis. Familial forms of ALS are caused by mutations in superoxide dismutase-1 (SOD1) gene, in 20% of cases. A therapeutic approach for SOD1-linked ALS is to suppress mutant SOD1 mRNA and protein in the affected tissues. We have previously shown that an adeno-associated virus serotype rh10 (AAV10) vector mediating exon skipping of the human SOD1 (hSOD1) pre-mRNA significantly reduces mutant SOD1 expression in SOD1G93A-ALS mice (Biferi et al., 2017). Viral particles were delivered through combined intra-cerebroventricular and systemic injections, efficiently reducing hSOD1 levels in MNs and SM. This approach prolonged survival and restored neuromuscular functions in vivo. In order to understand the tissue-specific contribution to the disease and to characterize the effects of the gene therapy approach in the human pathological context, we have generated in vitro models from patient's dermal fibroblasts (expressing SOD1 mutations: A4V, L144F and G93D). Primary fibroblasts were immortalized and trans-differentiated into myoblasts upon conditional MyoD over-expression, using established protocol. We analyzed SOD1 mRNA and protein expression under physiological condition or upon proteasome inhibition in fibroblasts and differentiated myotubes. We observed a statistical increase of hSOD1 mRNA and protein in patient cells relative to controls, after treatment with the proteasome inhibitor MG132. Proliferation and differentiation properties in myotubes were impaired in SOD1-mutant cells, with more striking effects in the A4V cell line (determining an aggressive disease progression in patients). We are currently testing the AAV-10 mediated gene therapy effects in these in vitro models. Our results will contribute to develop in vitro cellular models to investigate ALS physiopathology and gene therapy approaches.

Sirtuin 3 Inhibits Airway Epithelial Mitochondrial Oxidative Stress in Cigarette Smoke-Induced COPD.

Mitochondrial damage in airway epithelial cells plays an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Sirtuin 3 (Sirt3) is a mitochondrial deacetylase regulating mitochondrial function, but its role in the pathogenesis of COPD is still unknown. The aim of the present study was to investigate the effect of Sirt3 on airway epithelial mitochondria in cigarette smoke-induced COPD. Our present study has shown serious airway inflammation, alveolar space enlargement, and mitochondrial damage of the airway epithelium in COPD rats. Compared to the control rats, Sirt3 protein expression was significantly decreased in the airway epithelium and lung tissue homogenate from COPD rats. In airway epithelial cells (BEAS-2B), cigarette smoke extract (CSE) treatment significantly decreased mRNA and protein expression of Sirt3 and manganese superoxide dismutase (MnSOD), as well as MnSOD activity in a concentration and time-dependent manner. Sirt3 siRNA further significantly intensified the decreases in MnSOD expression and activity and aggravated mitochondrial oxidative stress and cell injury when airway epithelial cells were treated with 7.5% CSE. In contrast, Sirt3 overexpression significantly prevented the decrease of MnSOD expression and activity and improved mitochondrial oxidative stress and cell injury in CSE-treated airway epithelial cells. These data suggest that Sirt3 inhibits airway epithelial mitochondrial oxidative stress possibly through the regulation of MnSOD, thereby contributing to the pathogenesis of COPD.

Astragaloside IV Extends Lifespan of Caenorhabditis elegans by Improving Age-Related Functional Declines and Triggering Antioxidant Responses.

Astragaloside IV (AS-IV) is a representative component of astragaloside saponins in dried roots of Astragali Radix. Astragaloside possesses a broad spectrum of pharmacological activities, including antibacterial, anti-fibrosis, antioxidant, anti-inflammatory, and neuroprotective effects. However, the role of AS-IV in antiaging remains unclear. In this article, we studied the function of AS-IV in antiaging by using the Caenorhabditis elegans (C. elegans) model. We showed that AS-IV can prolong the lifespan of C. elegans in a natural aging model, a paraquat injury model, and a heat stress model and improve the movement capacity of nematodes. 1H-NMR data indicate an improvement of glutamate content and a decrease in glucose in the AS-IV treatment group compared with the control. Further investigation revealed that AS-IV can induce the mRNA expression of superoxide dismutase (SOD) and catalase (CAT) genes and increase the activities of SOD and CAT in the nematode. Interestingly, AS-IV could not extend the lifespan of sod-1, sod-2, sod-3, sod-4, sod-5, ctl-1, ctl-2, ctl-3, and daf-16 mutants. These data indicate that AS-IV prevents aging via mainly improving age-related functional declines, the antioxidant capacity of nematodes and partially modulating the insulin/insulin growth factor 1 signaling pathway activity. Our results provide new insights into how AS-IV prevents and treats aging.

Therapeutic effect of phycocyanin on acute liver oxidative damage caused by X-ray

1) BackgroundPhycocyanin (PC) is a type of natural protein in algae with antioxidant and anti-inflammatory properties. However, the protective effect of PC on hepatic damage induced by X-ray remains unknown. 2) MethodsMale C57BL/6 mice were gavaged with 200mg/kg PC for consecutive 7 days before or after radiation. The blood samples and tissues were collected on days 1 and 7 after radiation exposure. 3) ResultsPretreatment or treatment with PC decreased significantly the levels of alanine aminotransferase (ALT), aspartate aminotransferase(AST) in the plasma. Histological evaluation further confirmed the protection of PC against radiation-induced hepatotoxicity. PC-treatment also increased the relative mRNA expression of superoxide dismutase (SOD) and glutathione (GSH-PX), and descended the ROS in the liver. Moreover, the expression of H2AX, an indicator of DNA damage in mice, of the PC-intervention group was much smaller than that of the radiation group. In vivo, PC-treatment markedly up-regulated NF-E2-related factor 2(Nrf2) expression and downstream gene such as hemeoxygenase-1 (HO-1), NQO1. 4) Conclusion: PC could attenuate the radiation-induced oxidative stress damage by activating Nrf2/ HO-1 signaling pathway, and reduce the radiation-induced DNA damage. Therefore, PC is a protective agent against radiation-induced liver damage.

Ascorbic Acid and Alpha-Tocopherol Contribute to the Therapy of Polycystic Ovarian Syndrome in Mouse Models.

Polycystic ovary syndrome (PCOS) affects up to 10% of women within reproductive ages and has been a cause of infertility and poor quality of life. Alteration in the oxidant-antioxidant profile occurs in PCOS. This study, therefore, investigates the contribution of ascorbic acid (AA) and alpha-tocopherol(ATE) on different PCOS parameters. The mifepristone and letrozole models were used, and young mature female mice were randomly assigned to groups of six per group. On PCOS induction with either mifepristone or letrozole, mice were administered AA and ATE at doses ranging from 10-1000mg/kg to 0.1-1000mg/kg in the respective models. Vaginal cytology, body weights, and temperature, as well as blood glucose, testosterone, and insulin levels, were measured. Total antioxidant capacity and malondialdehyde levels were analyzed. Determination of gene expression of some reactive oxygen species and histomorphological analysis on the ovaries and uteri were performed. At the end of the experiments, AA and ATE restored reproductive cycling, with AA being more effective. AA and ATE increased fasting blood glucose but had no significant effect on serum insulin levels. AA decreased testosterone levels, but ATE caused slight increases. AA and ATE both increased total antioxidant capacity and decreased malondialdehyde levels. AA and ATE also slightly upregulated the mRNA expressions of catalase, superoxide dismutase, and heme oxygenase 1 mainly. AA and ATE also decreased ovarian weight and mostly resolved cysts in the ovaries and congestion in the uterus. This study has shown that AA and ATE are beneficial in the therapy of PCOS.

Berberine mitigates oxidative damage associated with testicular impairment following mercury chloride intoxication.

In this study, we assessed the protective effect of berberine (BBR) against mercuric chloride (HgCl2)-induced reproductive impairment. Twenty-eight adult male Wistar albino rats were placed into four equal groups: control, BBR, HgCl2, and BBR+HgCl2. All the treatments were orally administered for seven consecutive days. The rats exposed to HgCl2 showed a marked decrease in testosterone accompanied by an increase in lipid peroxidation (LPO) and nitric oxide (NO). Additionally, HgCl2 decreased glutathione (GSH) content, deactivated catalase (CAT) and glutathione reductase (GR), and suppressed the activities and mRNA expression of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in the testicular tissue. In addition, histological deformations and testicular cell loss were recorded, as evidenced by the upregulation of caspase-3 following HgCl2 intoxication. Notably, BBR administration reversed the testicular impairments associated with HgCl2 exposure. These findings suggest that BBR protected the testicular tissue following HgCl2 exposure through inhibiting the disturbance in the testosterone level and enhanced the antioxidant capacity. PRACTICAL APPLICATIONS: Heavy metals are naturally existing metallic elements in the earth's crust. These trace metals have the potential to cause multiple adverse reactions to the living systems, even at low exposure doses. Human exposure may also result from industrial, agricultural, and domestic activities. Berberine (BBR, a naturally occurring quaternary benzylisoquinoline alkaloid) is a protoberberine and is the representative main active ingredient in all parts of Berberis species. BBR has been used widely in folk medicine worldwide for its multiple pharmacological and therapeutic activities; for example, in the treatment of digestive and reproductive system disorders, microorganism infections, hemorrhage and wound healing, and cardiovascular and ophthalmic diseases. We found that BBR administration was able to abrogate significantly the reproductive toxicity associated with Hg intoxication. This protective effect comes from its strong antioxidant, anti-inflammatory, and antiapoptotic activities; suggesting that BBR may be applied to alleviate reproductive toxicity associated with Hg intoxication.

Direct-acting antivirals restore systemic redox homeostasis in chronic HCV patients

Chronic hepatitis C therapy has completely changed in the last years due to the availability of direct-acting antivirals (DAAs). Removing the virus may be not enough since chronic infection deeply modifies immune system and cellular metabolism along decades of inflammation. Oxidative stress plays a significant role in maintaining systemic inflammation during chronic HCV infection. Other than removing the virus, effective therapy could counteract oxidative stress. This study investigated the impact of DAA treatment on circulating markers of oxidative stress and antioxidant defence in a cohort of patients affected by chronic hepatitis C. To this, an observational study on 196 patients who started therapy with DAA for HCV-related hepatitis was performed. Patients were assessed at baseline, 4 weeks after the initiation of therapy (4wks), at the end of treatment (EoT), and 12 weeks after the EoT (SVR12). Circulating oxidative stress was determined by measuring serum hydroxynonenal (HNE)- and malondialdehyde (MDA)-protein adducts, and 8-hydroxydeoxyguanosine (8-OHdG). Antioxidant status was evaluated by measuring the enzymatic activity and mRNA expression of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in peripheral blood mononuclear cells. We observed a reduction of serum 8-OHdG at 4wks, while the circulating level of both HNE- and MDA-protein adducts diminished at EoT; all these markers persisted low at SVR12. On the other side, we reported an increase in the enzymatic activity of all the antioxidant enzymes in PBMC at EoT and SVR12. Taking into account circulating 8-OHdG and antioxidant enzyme activities, patients with high fibrosis stage were those that had the most benefit from DAA therapy. To conclude, this study indicates that treatment with DAAs improves the circulating redox status of patients affected by chronic hepatitis C. This positive impact of DAA therapy may be related to its effectiveness on cutting down viremia and pro-inflammatory markers.

Ameliorative effects of histidine on oxidative stress, tumor necrosis factor alpha (TNF-α), and renal histological alterations in streptozotocin/nicotinamide-induced type 2 diabetic rats.

Objective(s):The present study sought to evaluate the beneficial effects of histidine (His) on oxidative stress, tumor necrosis factor alpha (TNF-α), renal histological alterations and anti-oxidant enzymes gene expressions in type 2 diabetic rats.Materials and Methods:Streptozotocin/nicotinamide (STZ/NA) induced diabetic rats were used as an animal model of type 2 diabetes. One group of rats received daily His (1000 mg/l) in drinking water for 8 weeks, whereas other groups (control and untreated diabetic groups) received only water. Different parameters such as glucose, insulin, insulin resistance, lipid profile, cardiac risk ratios, renal functional markers, and oxidative stress were determined in all groups. Moreover, renal histological alterations, mRNA expressions of anti-oxidant enzymes, and TNF-α were evaluated in the rats.Results:His exhibited a protective effect on glucose, insulin, insulin resistance, lipid profile, cardiac risk ratios, renal functional markers, oxidative stress, and TNF-α. Furthermore, His restored the renal histological alterations and normalized the augmented mRNA expressions of renal anti-oxidant enzymes (glutathione peroxidase (GPX) and Cu-Zn superoxide dismutase (Cu-Zn SOD)) and TNF-α.Conclusion:His could ameliorate diabetes complications related to oxidative stress, inflammation, dyslipidemia, hyperglycemia, insulin resistance, and nephropathy. Hence, the use of this amino acid is recommended for diabetic patients in order to reduce diabetes complications.

Modulated Expression and Activities of Ruditapes philippinarum Enzymes After Oxidative Stress Induced by Aerial Exposure and Reimmersion.

Ruditapes philippinarum, is an economically and scientifically important bivalve mollusk. Its tolerance of aerial exposure has long been considered an important trait for its survival under acute environmental stress. In this study, the effects of air exposure at high and low temperatures (28 and 4°C) on the survival, antioxidant and metabolic enzyme activities, and the expression of antioxidant and immune-related genes in R. philippinarum were investigated. The activities of antioxidant and metabolic enzymes [superoxide dismutase (SOD), α-amylase, and proline hydroxylase (PHD)] were significantly affected by aerial exposure and reimmersion (reoxygenation) at both low (4°C) and high (28°C) temperatures. Moreover, the mRNA expression of α-amylase, SOD, and C-type lectin was also examined, which reveals these genes were significantly affected by aerial exposure challenge. In addition, the effects of aerial exposure and reimmersion on survival rate were calculated to evaluate the recovery capacity of Manila clam after aerial exposure at high and low temperatures. All individuals survived under low temperature aerial exposure for 24 h and reimmersion for 120 h. However, individuals died after reimmersion for 12 h following high temperature aerial exposure, and mortality peak occurred at 48 h. These data indicate that long-term aerial exposure during the transportation of clams should be in a low temperature environment. This study demonstrates that enzyme expression and activities linked to the stress response increase during the aerial exposure of R. philippinarum and provide useful information for future work on the molecular basis of tolerance of aerial exposure stress.

Effects of starvation and subsequent re-feeding on intestinal microbiota, and metabolic responses in Nile tilapia, Oreochromis niloticus

The effects of starvation and re-feeding on Nile tilapia (Oreochromis niloticus) remain incompletely understood so far. In this study, the changes of intestinal microbiota, the metabolic and immune (antioxidant) responses of Nile tilapia (Oreochromis niloticus) during starvation and re-feeding stages were investigated. The metabolic responses (cortisol, ALT, AST, and leptin) and immune responses (hsp70, SOD, and CAT) were measured. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the starvation group significantly increased during starvation and decrease after re-feeding. Cortisol levels increase during starvation and decreased after re-feeding. During starvation, the mRNA expression of superoxide dismutase (SOD), catalase (CAT), and heat shock protein (hsp70) were up-regulated while leptin was down-regulated. The microbial taxonomic composition of the starvation group slightly increased from the feeding and re-feeding groups. The relative abundance of the top five bacterial phyla in the intestinal of Nile tilapia (feeding, starvation and re-feeding groups) were Fusobacteria, Proteobacteria, Firmicutes, Bacteriodetes, and Actinobacteria. The relative abundance of the top three intestinal bacterial genera of Nile tilapia (feeding, starvation and re-feeding groups) were Cetobacterium, Romboutsia, Plesiomonas, Bacteroides, and Clostridium senso stricto. Taken together, starvation and re-feeding can modulate the diversity of microbial communities, the metabolic and immune responses of Nile tilapia.

Evaluation of the effective dose of amygdalin for the improvement of antioxidant gene expression and suppression of oxidative damage in mice.

BackgroundLittle is known regarding the toxic and therapeutic doses of amygdalin. Treatment regimens and schedules can vary between humans and animal models, and there have been reports of cyanide toxicity due to amygdalin use.ObjectiveThe aim of this study was to evaluate the effect of different doses of amygdalin on antioxidant gene expression and suppression of oxidative damage in mice.MethodsForty adult male mice were divided randomly into four groups (n = 10) as follows and treated orally for two weeks: a control group treated with saline solution, a group treated with amygdalin at 200 mg/kg body weight, a group treated with amygdalin at 100 mg/kg body weight, and a group treated with amygdalin at 50 mg/kg body weight. Liver and testis samples were collected for gene expression, biochemical and histopathological analyses.ResultsThe mice treated with medium-dose amygdalin (100 mg/kg) showed upregulated mRNA expression of glutathione peroxidase (P < 0.01) and superoxide dismutase (P < 0.05) and significantly decreased lipid peroxidation (P < 0.05) in hepatic and testicular tissues compared to those in the untreated groups (controls), with mild histopathological effects. The mice treated with high-dose of amygdalin (200 mg/kg) showed downregulated mRNA expression of glutathione peroxidase and superoxide dismutase (P < 0.01) and significantly increased lipid peroxidation (P < 0.05) in both hepatic and testicular tissues compared to those in the untreated groups (controls), with an apparent effect at the histopathological level. No effects were observed in the mice treated with low-dose amygdalin (50 mg/kg) at the gene, protein and histopathological level.ConclusionLow-and medium-dose amygdalin did not induce toxicity in the hepatic and testicular tissues of male mice, unlike high-dose amygdalin, which had a negative effect on oxidative balance in mice. Therefore, amygdalin at a moderate dose may improve oxidative balance in mice.

Dietary ginger improve growth performance, blood parameters, antioxidant capacity and gene expression in Cyprinus carpio

The present study evaluates the effect of dietary ginger (Zingiber officinale) on growth performance, haematological parameters, antioxidant status of Common carp (Cyprinus carpio). Fish with initial weight of 14±1.95 g were allocated into twelve tanks at density of 18 fish per tank.Fish were fed four experimental diets containing 0 (as control diet), 0.5, 2 and 5% ginger powder for 56 days. After 56 days of culture, fish fed 2 and 5% ginger diets showed significantly increase final weight, weight gain, specific growth rate (SGR) and feed conversion ratio (FCR) (p<0.05) compared to the control diet. Significantly higher the number of leucocytes (WBC), erythrocytes (RBC), Haematocrit, lymphocyte, monocyte and neutroplil levels were found in the fish fed 2 and 5% ginger diets compared to the control diet. Superoxide dismutase (SOD), catalase (CAT) and Glutathione peroxidase (GPX) enzyme activities and Malondialdehyde (MDA) level were significantly higher in the groups fed 2 and 5% ginger diets (p<0.05). SOD, CAT and GPX mRNA expression levels of fish fed 2 and 5% ginger diets were significantly down-regulated compared to the control diet (p<0.05). The results suggested that dietary ginger can be improve growth performance, health status and antioxidant capacity of Common carp

Improvement of Flavonoids in Lemon Seeds on Oxidative Damage of Human Embryonic Kidney 293T Cells Induced by H2O2.

In this study, flavonoids in lemon seeds (FLS) were used to assess its improvement on the oxidative damage of human embryonic kidney 293T cells (HEK 293T cells) induced by H2O2. In vitro experiments showed that the survival rates of HEK 293T cells treated with different flavonoid concentrations (50 μg/mL, 100 μg/mL, and 150 μg/mL) exceeded 95%, indicating no significant toxic effect. Compared with the normal group, H2O2 (0.3 mmol/L) resulted significantly in oxidative stress injury of HEK 293T cells. The survival rate of the damaged cells increased after treatment with flavonoids, and the survival rate of cells treated with a high concentration (150 μg/mL) of flavonoids was 76.2%. Flavonoids also effectively inhibited H2O2-induced apoptosis. At the same time, flavonoid treatment significantly reduced the malondialdehyde content in cells and increased the levels of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GSH-Px). Quantitative polymerase chain reaction (qPCR) and Western blot analysis also suggested that FLS upregulated mRNA and protein expressions of CAT, SOD (SOD1, SOD2), GSH (GSH1), and GSH-Px in H2O2-induced oxidative damage of HEK 293T cells. The high-performance liquid chromatography analysis demonstrated that FLS contained six compounds, including gallocatechin, caffeic acid, epicatechin, vitexin, quercetin, and hesperidin. FLS were proven to have a good antioxidant capacity in vitro and improve significantly the oxidative damage of HEK 293T cells induced by H2O2. The biological activity value warrants investigation in additional studies.

Blood Pressure, Vascular Reactivity, and SOD Expression/Activity in Mas1 Receptor Knockout Rats in the Dahl Salt‐Sensitive Genetic Background

Dahl salt‐sensitive (SS) rats are the most widely used rodent genetic model of human salt‐sensitive hypertension. Earlier studies have established the importance of impaired regulation of the renin gene in contributing to endothelial dysfunction in SS rats and the protective effect of low dose angiotensin II (ANG II) infusion in restoring endothelial function via the ANG II type1 receptor (AT1R) in the SS genetic background. Activation of the Mas1 receptor (Mas1R) for angiotensin (1‐7) [ANG (1‐7)] generally opposes ANG II‐mediated effects of AT1R activation. In Sprague‐Dawley rats fed high salt diet, binding of ANG (1‐7) to the Mas1R can activate NRF2 defenses and restore impaired endothelial function. The goal of the present study was to provide initial phenotyping of a Mas1R knockout (Mas1R KO) rat in the Dahl SS genetic background. Mas1R KO rats and wild type (WT) controls (12–15 weeks old) were maintained on a low salt (LS; 0.4% NaCl) or switched to a high salt (HS; 4% NaCl) diet for 3 days or 10 days. Arterial pressure measured via telemetry was similar in Mas1R KO rats (121±7 mmHg, n=3) and WT rats fed LS diet (124 mmHg, n=2) and increased in WT (138 mmHg, n=2) and Mas1R KO rats (132±5 mmHg, n=3) fed HS diet for 10 days. In rats fed LS diet, Mas1R KO eliminated vasodilator responses to ANG (1‐7) and AVE0991 (Mas receptor agonist) in middle cerebral arteries (MCA) and mesenteric resistance arteries (MRA) and reduced acetylcholine (ACh) reactivity in MCA. Because superoxide dismutase (SOD) enzymes contribute to antioxidant defense by catalyzing the dismutation of superoxide radicals and help to prevent diseases related to oxidative stress and endothelial dysfunction, we measured superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2) activity in the liver of WT and Mas1R KO rats after 3 days of HS diet. High salt diet increased SOD1 activity (WT‐LS 1.04±0.16 U/ml vs WT‐HS 1.65±0.12 U/ml) and SOD2 activity (WT‐LS 0.76±0.03 U/ml vs WT‐HS 1.06±0.11 U/ml) in WT rats, but not in Mas1R KO rats. SOD1 and SOD2 mRNA expression (evaluated via RT‐PCR) tended to increase in WT rats fed HS diet vs. LS diet (1.3 – 1.8 fold change, p &gt; 0.05). SOD1 mRNA expression was significantly higher in HS‐fed Mas1R KO rats vs. Mas1R KO rats fed LS diet (2.8 fold change, p = 0.01) and SOD2 mRNA tended to increase as well (1.7 fold change; p = 0.1). SOD1 and SOD2 protein expression measured by Western blot analysis was similar in WT and Mas1R KO rats fed either LS or HS diet. The present study using the Mas1 receptor knockout in the Dahl SS rat genetic background suggests that Mas1R activation by its agonists (AVE 0991) and ANG (1‐7) improves endothelial function and may provide novel therapeutic strategies for the treatment of low‐renin salt‐sensitive hypertension.Support or Funding InformationNIH #HL‐128242 and NIH #R21‐OD024781