Superoxide Dismutase Production Research Articles

Activation of BKα channel provented uremic cardiomyopathy by attenuating oxidative stress

Background: Uremic cardiomyopathy and renal fibrosis contribute to chronic kidney disease (CKD)-induced morbidity and mortality. Our previous study found that activation of the large conductance calcium-activated potassium channels (BK channels) attenuated renal fibrosis in mice (Wang et al, Kid Int, 2021). We hypothesized that upregulation of BK channels would suppress cardiac fibrosis in CKD. Methods: CKD mice were induced by 5/6 nephrectomy. To upregulation of BKa channels, BMS-191011 (10 mg/kg BW) was given by IP injection every day for 6-8 weeks. Single channel recordings were used to analyze the activation of BKa channel. Blood pressure was measured by non-invasive photoplethysmography using the tail-cuff method. Echocardiogram was used to assess cardiac function. Cultured H9C2 cardiac myoblasts were used to detect the impact of BK opener (NS1916 20mM) on fibrosis markers and oxidative stress. The mRNA expression of BKa in heart was assayed by qPCR. The Hydrogen peroxide (H2O2) was tested by Amplex Red Hydrogen Peroxide Kit. DHE (Dihydroethidium) was assayed to detect superoxide. Superoxide Dismutase (SOD), a defense of oxidative enzyme, was measured using colorimetric activity kit. Results: The expression of BKa mRNA and protein were decreased and cardiac fibrosis was increased in the heart of CKD mice. Single channel recordings showed that human uremic serum (User) attenuated the activation of BKa channel. Upregulation of BKa channel by BSM-191011 decreased the CKD-induced increase in systolic and diastolic blood pressure. Echocardiogram showed that LV end-diastolic dimension increased in 5/6Nx mice, but back to normal after BK opener treatment. Ejection fraction also improved by this treatment. Fibronectin and vimentin were also significantly increased in CKD heart, whereas BMS treatment attenuated the amount of both proteins. In cultured cardiac myoblasts, User and TGFβ increased fibronectin and collagen I, which were in a dose-dependent manner. BKa openers (NS1916) limited the User and TGF-induced upregulation of both proteins in H9C2 cells. User also significantly increased H2O2 and superoxide production in cultured H9C2 cells. Activation of BKa significantly abolished this upregulation. In addition, User and TGFβ decrease SOD production, NS1916 blocked this decline in a dose dependent manner. Conclusions: Activation of BK channel in CKD animals attenuates cardiac fibrosis likely through reduction of uremia-induced oxidative stress. This study could provide new approaches for developing therapeutic strategies for treating uremic cardiomyopathy in chronic kidney diseases. the Department of Veteran Affairs MERIT Award 5I01BX000994 to HC. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Динаміка показників оксидантно-антиоксидантних реакцій у післяопераційному періоді у хворих на поширений перитоніт

Purpose - to determine the effect of the developed treatment of patients with peritonitis on the dynamics of indicators of oxidant-antioxidant reactions and to justify the need for the use of the proposed drugs in the complex treatment of peritonitis. Matherials and methods. The study included 124 patients with toxic and terminal stages of peritonitis, who were divided into 3 groups: the Group I included 39 patients in whose complex treatment we included cytochrome C; the Group II included 41 patients whose complex treatment included cytochrome C and a solution containing levocarnitine and arginine hydrochloride; the Comparison group included 44 patients in whom the specified drugs were not used. The content of diene conjugates, reactive oxygen species (ROS), S-nitrosothiols, superoxide dismutase (SOD), carbonyl groups, and sodium nitrite was determined in the patients. Statistical processing of the study results was performed using the Statistical software EZR v. 1.64. Results. In patients of the Group I and the Group II, the use of the proposed treatment contributed to the optimization of oxidant-antioxidant reactions: a decrease in the production of diene conjugates, ROS, S-nitrosothiols, carbonyl groups, sodium nitrite during the study, optimization of SOD production, especially in the Group II. In patients of the Comparison group, decompensation in oxidant-antioxidant reactions was determined, associated with a significant increase in the production of diene conjugates, ROS, S-nitrosothiols and a decrease in SOD during the entire period. These trends developed against the background of an increase in the content of nitric oxide products and the activation of processes associated with the oxidative modification of blood plasma proteins. Conclusions. The use of cytochrome C and a solution containing levocarnitine and arginine hydrochloride in the complex treatment of patients with disseminated peritonitis helps to optimize oxidant-antioxidant reactions, which leads to a decrease in the manifestations of inflammation. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Neuroprotective effects of galectin‑1 on cerebral ischemia/reperfusion injury by regulating oxidative stress.

Oxidative stress contributes to the pathology of cerebral ischemia/reperfusion (I/R) injury. Galectin-1 has shown an anti-oxidative stress effect. The present study investigated whether this anti-oxidative stress effect can account for the neuroprotective actions of galectin-1 induced by cerebral I/R injury. A cerebral I/R injury model was created in C57Bl/6 mice by transient occlusion of the middle cerebral artery, after which the mice were treated with galectin-1 for 3 days. Infarct volumes were measured. A rotarod test and neurological deficit score assessment was performed to evaluate the neurological deficits. Oxidative stress was evaluated by measuring the levels of reactive oxygen species (ROS) and lipid peroxidation malondialdehyde (MDA), while the anti-oxidative stress status was assessed by measuring molecules such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidation enzyme (GSH-Px) in the ischemic cerebral hemisphere of mice. The inflammatory cytokines, including Interleukin 1 (IL-1), IL-6 and tumor necrosis factor alpha (TNF-α) were measured, and the expression of microglia was evaluated by immunohistochemistry in the ischemic cerebral hemisphere of mice. Galectin-1 treatment ameliorated neurological deficits and reduced infarct volumes in the mice model with cerebral I/R injury. Moreover, it was demonstrated that galectin-1 can significantly alleviate cerebral I/R injury in the ischemic cerebral hemisphere by decreasing the production of ROS and MDA, but increasing the production of CAT, SOD and GSH-Px. Galectin-1 treatment decreased microglia expression, and IL-1, IL-6 and TNF-α levels in the ischemic cerebral hemisphere of mice. Galectin-1 could improve the outcome of cerebral I/R injury by alleviating oxidative stress. Moreover, the neuroprotective effect of galectin-1 in cerebral ischemia could be related to its anti-oxidative stress effect.

Enhancement of superoxide dismutase activity using mixed Lactobacillus casei and Saccharomyces cerevisiae cultures in simulated gastrointestinal conditions with encapsulation

Superoxide dismutase (SOD)-producing microorganisms have attracted increasing attention given their scavenge superoxide anions capabilities, but further strengthening methods are still urgently needed. Herein, Lactobacillus casei F18 and Saccharomyces cerevisiae J14 were isolated for their high SOD activity. Co-culturing these two strains emerged effective nutritional cooperation, which increased SOD activity by 198.22% and 66.35% compared to F18 and J14, respectively. SOD activity of co-culture “F18 +J14” was further enhanced by response surface methodology and metal ion stress. High-performance liquid chromatographic and RT-qPCR analyses of “F18 +J14” revealed that SOD production pathways were active and contents of metabolites acetate and ethanol were dramatically increased. Metabolite complementarity experiment also verified the addition of acetate and ethanol in vitro enhanced SOD activity and gene expression of “F18 +J14”. These findings highlighted co-culture and metal ion stress enhanced SOD activity by cross-stimulating cells with metabolites, affecting cell growth, and up-regulating SOD genes. Furthermore, “F18 +J14” based immobilized bio-beads were successfully synthesized with sodium alginate combined CaCO3, which exhibited the highest cell vitality and SOD activity in the simulated gastrointestinal conditions. Together, this study revealed the synergistic promotion mechanism of SOD biosynthesis by co-cultivation lactic acid bacteria and yeast, and shows great promise for management of oxidation stress related diseases.

Novel palladium(II) and Zinc(II) Schiff base complexes: Synthesis, biophysical studies, and anticancer activity investigation

BackgroundSchiff base metal complexes are considered promising chemotherapeutic agents due to their potential application in cancer therapy. MethodsThe current work sought to synthesize a brand-new Schiff base ligand obtained from 2-hydroxybenzohydrazide and (E)− 1-(2-(p-tolyl)hydrazono)propan-2-one with metal ions which included Pd(II) and Zn(II) ions. Elemental analyses, FT-IR, mass spectra, 1H NMR, UV-Vis spectrometer, and computational analysis characterized the compound's structure. In vitro, the breast cancer cell line (MCF-7) was tested for its sensitivity to Schiff base (HL) and its Pd(II) and Zn(II) complexes. The half-maximal inhibitory concentration IC50 of the compounds was determined and used to perform the comet assay, which was carried out to reveal the photo-induced DNA damaging ability of the compounds of individual cells. Moreover, the compounds' effects on antioxidant defense systems of enzymes in cells: superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities and oxidant Malondialdehyde (MDA) were examined in MCF-7 cells. ResultsThe Pd(II) complex displayed approximately the same IC50 as Cisplatin, while Zn(II) complex had better activity than Cisplatin with very low IC50, 1.40 μg/ml. Significant alterations in SOD, CAT, GPx, and MDA production were discovered, inducing oxidative stress, enlarging ROS production, and reducing the antioxidant amount. This change was approximately similar in most compounds. Consequently, it promoted apoptosis, particularly the Zn(II) complex, which demonstrated an improved impact because of its ability to influence the antioxidant defense systems of enzymes, mostly SOD and GPx, besides increasing MDA levels. ConclusionIt can be concluded that Zn(II) complex is the most effective anticancer drug since it induced a very similar genotoxic effect as Cisplatin and has a very low IC50 value.

Cardiopreventive effect of Klotho protein on oxidative stress, inflammation and apoptosis during cardiac ischemia/reperfusion injury

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Science Centre, Poland. Background The injury of the myocardium during ischemia/reperfusion (IRI) involves metabolic, morphological and contractile disorders [1]. Klotho is a membrane or soluble anti-aging protein. Recent studies have proven the correlation of Klotho deficiency and the occurrence and development of cardiovascular diseases [2]. Purpose This study aimed to evaluate the effect of Klotho protein supply on oxidative stress, inflammation and apoptosis in hearts subjected to IRI. Methods Isolated Wistar rat hearts perfused with the Langendorff method were subjected to ischemia, followed by reperfusion, in the presence or absence of Klotho protein. Hemodynamic parameters, and the levels of nitrate/nitrite (NOX)-, glutathione peroxidase (GPx), superoxide dismutase (SOD), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-Κb), nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), interleukin-6 (IL-6), matrix metalloproteinase 2 (MMP-2), cytochrome c (cyt c) and caspase-9 were evaluated. Results Infusion supply of Klotho during IRI resulted in a recovery of contractile function (p=0.031) and heart rate (p=0.02). Klotho protein contributed to increase in the production of GPx (p=0.045) and SOD (p=0.006). The levels of (NOX)- (p=0.022), NF-Κb (p=0.006), NFATc1 (p=0.027), IL-6 (p=0.002), MMP-2 (p=0.008), cyt c (p=0.0002) and caspase-9 (p=0.009) were decreased in the IRI+Klotho group, as compared to the IRI group. Conclusions Klotho protein preserved heart mechanical function, enhanced antioxidant capacity, and reduced inflammation and apoptosis in the injured heart. Thus, Klotho can be recognized as a novel cardiopreventive/cardioprotective agent in ischemic damage.

Enhancing the Productivity and Stability of Superoxide Dismutase from Saccharomyces cerevisiae TBRC657 and Its Application as a Free Radical Scavenger

Superoxide dismutase (SOD) is crucial antioxidant enzyme that plays a role in protecting cells against harmful reactive oxygen species (ROS) which are generated inside cells. Due to its functionality, SOD is used in many applications. In this study, Saccharomyces cerevisiae TBRC657 was selected as the SOD producer due to its high SOD production. After investigating an optimized medium, the major components were found to be molasses and yeast extract, which improved SOD production up to 3.97-fold compared to a synthetic medium. In addition, the optimized medium did not require any induction, which makes it suitable for applications in large-scale production. The SOD formulation was found to increase the stability of the conformational structure and prolong shelf-life. The results show that 1.0% (w/w) trehalose was the best additive, in giving the highest melting temperature by the DSF method and maintaining its activity at more than 80% after storage for 6 months. The obtained SOD was investigated for its cytotoxicity and ROS elimination against fibroblast cells. The results indicate that the SOD enhanced the proliferation and controlled ROS level inside the cells. Thus, the SOD obtained from S. cerevisiae TBRC657 cultured in the optimized medium could be a candidate for use as a ROS scavenger, which can be applied in many industries.

Carboxymethylated pachyman induces ferroptosis in ovarian cancer by suppressing NRF1/HO-1 signaling.

Carboxymethylated pachyman (CMP) is characterized by immune regulatory, antitumor and antioxidant activities. However, whether CMP contributes to the treatment of ovarian cancer has yet to be explored. The role of CMP in ovarian cancer cell death was analyzed using CCK-8 and flow cytometry assays. The data showed that CMP induced ovarian cancer cell death in a dose-dependent manner. Furthermore, CMP-induced cell death could be largely reversed by preincubation with ferrostatin-1 (Fer-1) but not 3-methyladenine or necrostatin-1. Reverse transcription-quantitative PCR analysis indicated that CMP significantly increased prostaglandin-endoperoxide synthase 2 (PTGS2) and Chac glutathione specific γ-glutamylcyclotransferase 1 (CHAC1) mRNA levels, but preincubation with Fer-1 obviously reduced PTGS2 and CHAC1 mRNA levels in SKOV3 and Hey cells. The intracellular levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) and Fe2+ were then quantified The data showed that 100 and 200 µg/ml CMP enhanced the production of SOD, MDA and Fe2+ but decreased GSH levels in SKOV3 and HEY cells. These data indicated that CMP could induce ferroptosis in ovarian cancer cells. More importantly, in vitro and in vivo studies indicated that CMP significantly suppressed nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), cystine/glutamate antiporter system X(c)(−) (xCT) and glutathione peroxidase 4 (GPX4) expression in ovarian cancer cells and tumors. In conclusion, the present study showed novel data that CMP could induce ferroptotic death in ovarian cancer cells by suppressing Nrf2/HO-1/xCT/GPX4. All these findings indicate that CMP may have great potential in anti-ovarian cancer cell therapy by inducing ferroptosis.

Effect of Aronia melanocarpa fruit juice on the antioxidant defense system in rats with diet-induced metabolic syndrome

INTRODUCTION: The role of oxidative stress in the development of metabolic syndrome (MS) has been well established. Aronia melanocarpa fruits are very rich in polyphenols, which possess an antioxidant effect. AIM: The current study aimed to evaluate the effect of Aronia melanocarpa fruit juice (AMFJ) on the antioxidant defense system in rats with diet-induced MS. MATERIALS AND METHODS: Fifty male Wistar rats were allocated into 5 groups: control, MS, MS + AMFJ 2.5 , MS + AMFJ 5 and MS + AMFJ 10. For 10 weeks,the control group received regular diet and the other groups—high-fat high-fructose diet (HFHF). During this period, the control group and MS group were treated daily orally with 10 mL/kg distilled water and the other groups—with increasing volume (2.5 mL/kg, 5 mL/kg, and 10 mL/kg) of AMFJ. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were measured in the serum using commercial colorimetric kits. RESULTS: A significantly higher SOD level was observed in MS group (0.0069 ± 0.0007 U/mL) compared to the control group (0.0051 ± 0.0003 U/mL) (p<0.01). AMFJ treatment returned the level of SOD to the control values, with the effect being significant in MS + AMFJ 2.5 (0.0053 ± 0.0003 U/mL)and MS + AMFJ 5 (0.0053±0.0003 U/mL) groups (p<0.05 vs. MS). No significant difference was detected in the activity of GPx in all groups. CONCLUSION: HFHF diet-induced MS might be associated with superoxide production and compensatory activation of SOD. Due to its antioxidant properties, AMFJ counteracted these processes in the treated groups. Neither the HFHF diet, nor the AMFJ treatment affected the activity of GPx.

POTENTIAL EFFECT OF THYME AND OREGANO ESSENTIAL OILS TO ANTIMICROBIAL ACTIVITY AND YEAST ENZYMATIC ANTIOXIDATIVE SYSTEM

This in vitro study was aimed to assess the effects of Thymus vulgaris L. and Origanum vulgare L. essential oils (EOs) on cell viability and enzymatic antioxidative system in yeast Saccharomyces cerevisiae. Yeasts were cultivated with different concentrations of thyme (Thymus vulgaris L.) (0.05 – 1.00 µl/ml) and oregano (Origanum vulgare L.) (0.05 – 0.70 µl/ml) EOs for 24 hours. For the present study, gas chromatography-mass spectrometry (GC-MS) analysis was used to quantify chemical composition of EOs. The main components of thyme essential oil were caryophyllene (7.94%), p-cymene (18.55%), and thymol (46.55%). In essential oil of oregano, there was found p-cymene (8.43%) and carvacrol (69.99%) mainly. The antimicrobial activity of selected EOs was investigated by the microbroth dilution method using 96 well microtiter plates. The most potent essential oil able to inhibit growth of Saccharomyces cerevisiae was oregano EO with the values of MIC50 0.468845 µl/ml and MIC90 0.501845 µl/ml. Also, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assessed. According to our results, Thymus vulgaris L. significantly increased the SOD levels at 0.2, 0.4, 0.5 to 0.9 µl/ml and GPx production at 0.05 to 0.7 µl/ml (P<0.001). In case of Origanum vulgare L., significant increase of SOD production at 0.05 - 0.2 µl/ml (P<0.001) was observed. This essential oil also elevated GPx concentration in all experimental groups (P<0.001) significantly. Data from this study suggest that selected EOs have significant effects on enzymatic antioxidants that may affect the first-line cell defense system against oxidative stress.

DNA Damage Detection after Chronic Exposure and Radio-adaptive Response of Naturally Occurring Radioactive Materials (NORM)

The objective of this study was to understand the effect of chronic low-dose radiation that induces in vivo radio-adaptive response. The animals were exposed chronically to natural radioactive materials (NORM) for one and two months which correspond to 10 &20 mSv which are comparable to the reality of areas with high background natural radiation (HBNR). Radioactivity assessment of samples was done using a high purity germanium -ray spectrometer. We conduct the study to understand the radio-adaptive response by exposure the experimental animals to challenge dose 2 Gy after continuous exposure to 10 & 20 mSv. Comet assay was performed in addition to some of antioxidant enzymes concentrations (superoxide dismutase (SOD), catalyze enzyme (CAT), reduced and oxidized glutathione (GSH). Chronic exposure to 10 and 20 mSv showed DNA damage as a high elevation in comet assay, parameters. Also the results of the present work showed a low production of antioxidant enzymes CAT, SOD & GSH in both chronic exposure (10 & 20 mSv and challange dose 2 Gy). The exposure to 20 mSv before 2Gy gamma rays resulted in an enhancement of antioxidant enzymes accompanied with decreasing in free radical which is represents as an adaptive response. . It can be said that for low radiation doses, which are less than 100 mSv, their effects differ from high doses, so that it can be said that the body has an adaptive response and the application of the linear non-threshold model must be reviewed for these doses.

N-docosahexaenoylethanolamine reduces neuroinflammation and cognitive impairment after mild traumatic brain injury in rats

At present, there is a growing interest in the study of the neurotropic activity of polyunsaturated fatty acids ethanolamides (N-acylethanolamines). N-docosahexaenoylethanolamine (DHEA, synaptamide) is an endogenous metabolite and structural analogue of anandamide, a widely studied endocannabinoid derived from arachidonic acid. The results of this study demonstrate that DHEA, when administered subcutaneously (10 mg/kg/day, 7 days), promotes cognitive recovery in rats subjected to mild traumatic brain injury (mTBI). In the cerebral cortex of experimental animals, we analyzed the dynamics of Iba-1-positive microglia activity changes and the expression of pro-inflammatory markers (IL1β, IL6, CD86). We used immortalized mouse microglial cells (SIM-A9) to assess the effects of DHEA on LPS-induced cytokines/ROS/NO/nitrite, as well as on CD206 (anti-inflammatory microglia) and the antioxidant enzyme superoxide dismutase (SOD) production. In vivo and in vitro experiments showed that DHEA: (1) improves indicators of anxiety and long-term memory; (2) inhibits the pro-inflammatory microglial cells activity; (3) decrease the level of pro-inflammatory cytokines/ROS/NO/nitrites; (4) increase CD206 and SOD production. In general, the results of this study indicate that DHEA has a complex effect on the neuroinflammation processes, which indicates its high therapeutic potential.

Combination of Scalp Acupuncture with Exercise Therapy Effectively Counteracts Ischemic Brain Injury in Rats

BackgroundStroke is one of the leading causes of death and disability worldwide. Scalp acupuncture and exercise therapy have been proven as two effective methods for the treatment of stroke. However, their combined action and mechanisms have not been fully elucidated. The present study aimed to investigate the protective effect of scalp acupuncture combined with exercise therapy on neurons in rats with ischemic brain injury. Methods100 rats were randomly divided into 5 groups including sham group, model group, acupuncture group, rehabilitation group, and experimental group (scalp acupuncture combined with exercise therapy). Middle cerebral artery occlusion (MCAO) model in rats was established according to Longa modified suture method to mimic ischemic stroke. The modified Bedexer's neurological function score was used to evaluate the neurological deficits of rats and the brain infarct volume was measured using 2, 3, 5-triphenyl tetrazolium chloride monohydrate (TTC) staining. Moreover, the apoptosis in the hippocampus was detected by western blotting and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The pro-inflammatory cytokines such as interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α), reactive oxygen species (ROS) and superoxide dismutase (SOD) were determined by corresponding kits. Immunohistochemistry or immunofluorescence was performed to detect the expression of brain-derived neurotrophic factor (BDNF), S100β and glial fibrillary acidic protein (GFAP) in the hippocampi of rats. ResultsThe neurological deficit score, the expression levels of apoptotic factors such as cleaved caspase-3 and Bax, and the TUNEL-positive cell rate of the experimental group were significantly lower than those of the acupuncture group and the rehabilitation group. However, apoptosis inhibitor Bcl-2 showed downregulated expression in the MCAO model rats but this trend was reverted by single and combinatorial treatments. In addition, the contents of TNF-α, IL-1β and ROS in the acupuncture group and the rehabilitation group were significantly lower than those in the model group, but higher than the experimental group. While the opposite results were obtained in SOD activity. Furthermore, compared with the model group, the ratios of BDNF, S100β, and GFAP-positive cells in the acupuncture, rehabilitation and experimental groups were significantly increased, and the highest ratios were recorded in the experimental group. ConclusionsThis study demonstrated that scalp acupuncture combined with exercise therapy effectively counteracts ischemic brain injury via the downregulation of pro-inflammatory mediators and ROS, the increased production of the antioxidant enzyme SOD, neurotrophic factor BDNF and astrocyte activities.

Oxidative stress-mediated mitochondrial pathway-dependent apoptosis is induced by silica nanoparticles in H9c2 cardiomyocytes

The use of silica nanoparticles (SiNPs) is increasing in popularity; however, the emissions released during manufacturing, use and during the disposal stages potentially harm the environment. SiNPs can enter the body and cause cardiac toxicity indirectly or directly. However, toxicological data on SiNPs in cardiac cells in vitro, and the detailed molecular mechanisms by which damage is caused remain unclear. In the present study, oxidative stress-mediated apoptosis and cytotoxicity induced by SiNPs in H9c2 cells were examined. H9c2 cells were used to explore the mechanisms of toxicity by treating cells with 0, 25, 50, 100, and 200 µg/ml SiNPs, with and without 3 mM of the reactive oxygen species (ROS) scavenger, N-acetyl-l-cysteine (NAC), for 24 h. The results showed that SiNPs decreased cell viability and proliferation by increasing the release of lactate dehydrogenase (LDH) and inducing apoptosis in H9c2 cells. ROS levels were significantly increased in a dose-dependent manner. Additionally, the levels of superoxide dismutase (SOD), glutathione (GSH), and GSH-peroxidase (Px) were significantly decreased following exposure to SiNPs. Treatment with NAC attenuated LDH release; the levels of ROS, SOD, GSH, and GSH-Px production were increased, and SiNPs-induced mitochondrial pathway-dependent apoptosis was reduced. These results demonstrate that apoptosis and cytotoxicity induced by SiNPs in H9c2 cells are a result of ROS-mediated oxidative stress. These data suggest that exposure to SiNPs is a potential risk factor for cardiovascular disease.

Helix B Surface Peptide Protects Cardiomyocytes From Hypoxia/Reoxygenation-induced Autophagy Through the PI3K/Akt Pathway.

Helix B surface peptide (HBSP) is a newly discovered tissue-protective erythropoietin derivative that provides benefits after myocardial ischemia/reperfusion. This study explores the cardioprotective effects of HBSP in myocardial cells in response to hypoxia/reoxygenation injury and its potential mechanism. In this study, rat ventricular (H9c2) cell cultures were established and pretreated with HBSP. H9c2 cardiomyocytes were randomly assigned to the control, H/R, H/R + LY294002 (a PI3K inhibitor), HBSP + H/R, and HBSP + H/R + LY294002 groups. The pretreated cardiomyocytes underwent H/R, and the cardiomyocytes were monitored for viability through a CCK-8 assay, whereas flow cytometry was used to test cell apoptosis. Orgotein Superoxide Dismutase (SOD) and lactate dehydrogenase (LDH) expression were monitored by SOD and LDH kits, respectively. The expression of LC3 autophagosomes was determined by immunocytochemistry. The expression of LC3II/LC3I, p-Mammalian Target of Rapamycin (mTOR) mTOR, mTOR, Beclin 1, p-PI3K, PI3K p-Akt, and Akt was determined by Western blotting. HBSP increased cell viability and reduced SOD and LDH production, and it also reduced H/R-induced cell apoptosis. Moreover, the expression of the autophagy-related proteins (LC3II/LC3I) was inhibited by HBSP, whereas the expression of p-PI3K, p-Akt, and p-mTOR was enhanced. However, the PI3K inhibitor (LY294002) notably abolished these effects in H9c2 cells. HBSP inhibits excessive autophagy and apoptosis induced by H/R by activating the PI3K/Akt pathway. HBSP may potentially be a therapeutic intervention for myocardial ischemia/reperfusion injury.

Three-weeks moderate aerobic exercise in increasing production of endogenous antioxidant enzyme and lowering oxidative stress level among sedentary men

Background: During exercise, there will be an increase of free radical due to the elevation of oxygen consumption to fulfill the increasing demand for energy. This free radical could produce by muscle and other tissue, especially from the leakage of mitochondria respiratory chain reaction. Moderate exercise training may provide adequate protection against exercise-induced oxidative stress by elevating the antioxidant level and healthier mitochondria that will produce less free radicals. This research will investigate the change of baseline level of MDA and superoxide dismutase (SOD) and also MDA and SOD level after a single bout of moderate exercise before and after three weeks of intervention.Methods: Seven sedentary men with age average 17.2 + 0.2 years recruited for this study. They were meet the criteria of healthy sedentary men. Moderate aerobic exercise of 30 minutes running at 11 – 13 RPE Borg Scale, three times a week for three weeks in a row was applied to the subject. MDA and SOD examination was done by chromatographic assays for total plasma level. The collected data were analyzed using dependent student t-test. Data were normally distributed.Results: The result of this research found a significant elevation of SOD production (p<0.01) after a single bout moderate exercise after three week training (19.98 + 15.52 vs 40.43 +19.64 mg/ml) and significant depression of MDA production (p>0.05) after training program (0.79 + 0.62 vs 1.97 + 0.94 nmol/ml).Conclusion: There is no significant change in post-exercise MDA and SOD level before and after the training program. Initial fitness status, food intake, lifestyle could affect baseline level of MDA and SOD before and after exercise.

Cerium Oxide Nanoparticles Reduce X-ray Irradiation-Induced Damage to the Immune Cells by Upregulation of Superoxide Dismutase and Glutathione Peroxidase

Ionizing radiation cause serious damage to humans by inducing reactive oxygen species (ROS). Immune system is one of the most sensitive system to ionizing radiation. Cerium oxide (CeO2) nanoparticles are strong ROS scavengers. In this research we observed that whether CeO2 nanoparticles could improve immune function of X-rays irradiated mice and their antioxidant potential. The whole body of mice were once exposed to 4 Gy X-ray and treated with 5 nm CeO2 nanoparticles. Then, immune cells distribution in peripheral blood (PB), spleen T cells proliferation, spleen and thymus indexes, the concentration of Glutathione Peroxidase (GSH-Px) and the activity of Superoxide Dismutase (SOD) were analyzed. The results showed that CeO2 nanoparticles could inprove immune cells distribution in PB and increase spleen T cells proliferation, thymus and spleen indexes, concentration of GSH-Px and activity of SOD in irradiated mice. The results suggest that CeO2 nanoparticles can improve immune function by inducing the production of GSH-Px and SOD in X-ray irradiated mice.

Superoxide dismutase response and the underlying molecular mechanism induced by iodoacetic acid

Given the ubiquity of iodinated disinfection by-products (I-DBPs) in drinking water and their prominent toxicity, it is of vital significance to evaluate I-DBPs toxicity and explore the underlying mechanism. The toxicity of iodoacetic acid (IAA), a typical type of I-DBPs, might be linked with oxidative stress. However, it remains unknown for the response of antioxidant enzyme superoxide dismutase (SOD) in the mouse primary hepatocytes when exposed to IAA and the underlying mechanism. This study explored SOD response to IAA and the underlying mechanisms at the molecular and cellular levels. Under IAA exposure, the observed increase of SOD activity in the hepatocytes was caused by the increase of SOD production via ROS stimulation and the increase of SOD molecular activity. Molecular experiments showed that IAA binds to SOD molecule mainly via electrostatic forces with one binding site around the active site and six binding sites in the surface of protein. The binding interaction leads to the conformational changes of SOD and the disruption of protein aggregates. This work could offer basic data for the comprehensive understanding of the adverse effects of IAA and references for assessing the harmful effects of DBPs.

Effect of euphorbia factor L1 on oxidative stress, apoptosis, and autophagy in human gastric epithelial cells

BackgroundEuphorbia factor L1 (EFL1), a lathyrane-type diterpenoid from the medicinal herb Euphorbia lathyris L. (Euphorbiaceae), has been reported for many decades to induce gastric irritation, but the underlying mechanisms remain unclear. PurposeThe objective of this study was to investigate EFL1-induced cytotoxicity and the potential mechanisms of action on the human normal gastric epithelial cell GES-1. MethodsGES-1 cells were treated with EFL1 (12.5–200 μM) for different time intervals, and cell survival, LDH release, intracellular reactive oxygen species (ROS), malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity were detected. Mitochondrial membrane potential (MMP) assay, DAPI staining, DNA fragment assay, and annexin V-FITC/PI staining were performed. The interaction between EFL1 and Bcl-2, cytochrome c, caspase-9, caspase-3, PI3K, AKT, and mTOR proteins was simulated by molecular docking. The mRNA and protein expression of apoptosis and autophagy factors were detected by RT-qPCR and Western blotting. ResultsEFL1 decreased survival, increased LDH leakage, and induced abnormal production of ROS, MDA and SOD in GES-1 cells. Mitochondria-mediated apoptosis was characterized by decreased MMP, condensed nuclei, fragmented DNA, and increased apoptosis rate. EFL1 interacted with proteins via hydrogen bonding. The mRNA, total or phosphorylated protein expression of Bcl-2, mitochondrial cytochrome c, PI3K, AKT, mTOR and p62 were downregulated; in contrast, those of cytoplasmic cytochrome c, cleaved caspase-9, cleaved caspase-3, LC3-ll and Beclin-1 were upregulated. ConclusionThese findings indicated that EFL1 decreased the survival of GES-1 cells through EFL1-induced oxidative stress, activation of the mitochondria-mediated apoptosis as well as autophagy via inhibition of the PI3K/AKT/mTOR pathway.

Delivery of Superoxide Dismutase Using Cys2-His2 Zinc-Finger Proteins.

Therapeutic proteins have shown great potential in treating life-threatening diseases, but the hydrophilicity and high molecular weight hamper their passing through the cell membrane. Cell-penetrating peptide (CPP)-assisted protein delivery is a simple and efficacious strategy to promote the cellular uptake of therapeutic proteins. We recently demonstrated that the engineered Cys2-His2 zinc-finger domains possess intrinsic cell permeability, which could be leveraged for intracellular protein delivery. Here we applied this method to deliver superoxide dismutase (SOD), a therapeutic protein widely used in preclinical and clinical studies. We present a protocol for the production and delivery of zinc-finger domain-fused SOD. This protocol can be extended for delivering other therapeutic proteins.