Release Of Superoxide Dismutase Research Articles

Ammidin ameliorates myocardial hypoxia/reoxygenation injury by inhibiting the ACSL4/AMPK/mTOR-mediated ferroptosis pathway

ObjectiveThe aim of the present study was to investigate the therapeutic effect of ammidin on hypoxia/reoxygenation (H/R) injury in primary neonatal rat cardiomyocytes by observing the role of ferroptosis in the process of H/R injury, and to verify its target and regulatory signaling pathways.MethodsThe network pharmacology analysis was used to predict the biological processes, core targets and related signaling pathways of Angelica dahurica in the treatment of ferroptosis. Cell viability was assessed using live cell imaging and cell counting kit-8. Lactate dehydrogenase (LDH), reactive oxygen species (ROS) production, and malondialdehyde (MDA), superoxide dismutase (SOD) and mitochondrial membrane potential (MMP) content were determined to assess the level of ferroptosis. Western blotting was performed to measure protein expression.ResultsNetwork pharmacology predicted that Acyl-CoA synthetase long chain family member 4 (ACSL4) was highly associated with myocardial H/R injury in the intersection of Angelica dahurica and ferroptosis. The top three active components of Angelica dahurica were found to be mandenol, alloisoimperatorin and ammidin, among which ammidin was found to have the strongest binding to the target proteins of the ACSL4/AMPK/mTOR pathway. H/R reduced the viability of cardiomyocytes, while the inhibition of ferroptosis by ferrostatin-1 alleviated the H/R-induced inhibition of cardiomyocyte viability. This was evidenced by the increased cell viability, SOD release, MMP level and glutathione peroxidase 4 (GPX4) protein expression, as well as the decreased LDH and MDA release and ROS production and ACSL4 protein expression (P < 0.05). To verify the existence of ferroptosis in myocardial hypoxia/reoxygenation injury. In addition, ammidin increased cell viability and GPX4 protein expression (P < 0.05), decreased ROS generation, and MDA and MTT expression (P < 0.05), then inhibited ferroptosis, and finally alleviated myocardial H/R injury by regulating the ACSL4/AMPK signaling pathway.ConclusionsNetwork pharmacology was used to predict the correlation between ammidin and ferroptosis following myocardial H/R injury. It was demonstrated that ammidin may regulate ferroptosis by inhibiting the ACSL4/AMPK/mTOR signaling pathway and reduce H/R injury in cardiomyocytes.

Ruscogenin regulates endogenous antioxidation in dopamine neurons by activating Keap1/Nrf2/HO-1 pathway

Purpose: To investigate the effect of ruscogenin (RUS) on cell viability, lipid peroxidation and mitochondrial dysfunction in a Parkinson’s disease (PD) model.Methods: The neuroblastoma cell line SH-SY5Y was modified with 1-methyl-4-phenylpyridine (MPP+) to establish a PD model. RUS (1 or 10 μM) was used to treat MPP+ induced SH-SY5Y cells. Cell viability was assessed using CCK-8 assay. The concentrations of lactate dehydrogenase (LDH), superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) and glutathione peroxidase (GPx) were determined by enzyme-linked immunosorbent assay (ELISA). ATP production, Ca2+ concentration and JC-1 were quantified using commercial kits. The expression levels of tyrosine hydroxylase (TH), Keap1, Nrf2 and HO-1 were evaluated by western blot analysis.Results: RUS protected cell viability, reduced LDH production, and elevated TH expression in MPP+- modified SH-SY5Y cells. RUS promoted the release of SOD, CAT and GPx, but suppressed MDA production. Furthermore, RUS enhanced ATP metabolism, decreased Ca2+ leakage and maintained mitochondrial function. RUS also repressed Keap1 expression but increased Nrf2 and HO-1 levels.Conclusion: RUS enhances cell viability while alleviating cytotoxicity, lipid peroxidation and mitochondrial dysfunction in dopamine neurons through the activation of Keap1/Nrf2/HO-1 signaling pathway. Thus, RUS is a promising therapeutic candidate for PD treatment.

Encapsulation of SOD in chitosan-coated gel particles of alginate or mixture of alginate and shellac for targeted intestinal delivery

The chitosan-coated alginate gel particles (CS/SA) and chitosan-coated alginate-shellac gel particles (CS/SA/Lac) were constructed to serve as a potential oral delivery system to enhance the stability and activity of superoxide dismutase (SOD) in this study. The encapsulation efficiency (EE), enzyme activity retention ratio (ER), particle structure, and SOD release behavior, as well as the effect of shellac, were evaluated. The best formula is determined by the percentage of total effective activity (PTEA).The results showed that CS/SA prepared by 0.30‰ chitosan, 1.00% CaCl2, 2.00% sodium alginate (M/G = 1:1, Mw: 38,857 kDa), and gelation solution pH 5.50 had the best chitosan shell, the strongest gel structure and the maximum PTEA which was 10.75 ± 1.45%. Based on the optimal formula, addition of shellac that formed shellac-Ca2+ microspheres in the gel network was averse to SOD encapsulation, but it significantly improved the stability of SOD in gastric conditions. By adding 15.00% shellac, the PTEA reached 16.30 ± 1.64%. At optimal formulation, SOD encapsulated in CS/SA was completely released in the stimulated intestinal fluid and accumulative release enzyme activity (EA) was 966.62 ± 103.75 (U) after 6 h. However, the CS/SA/Lac with a low swelling ratio only released SOD in stimulated colonic fluid and the EA value reached 1692.28 ± 101.82 (U) after 1 h. These findings demonstrated that CS/SA and CS/SA/Lac were excellent candidates for SOD oral administration, which was useful in the food and biomedical industries for developing functional products containing SOD.

Novel organic selenium source hydroxy-selenomethionine counteracts the blood-milk barrier disruption and inflammatory response of mice under heat stress.

Heat stress (HS) in summer has caused huge economic losses to animal husbandry production recently. When mammary gland is exposed to high temperatures, it will cause blood-milk barrier damage. Hydroxy-selenomethionine (HMSeBA) is a new selenium source with better guarantee of animals' production performance under stress, but whether it has protective effect on heat stress-induced blood-milk damage is still unclear. We established mammary epithelial cells and mice heat stress injury models to fill this research gap, and hope to provide theoretical basis for using HMSeBA to alleviate heat stress damage mammary gland. The results showed that (1) Heat stress significantly decreases in vitro transepithelial electrical resistance (TEER) and cell viability (P < 0.01), and significantly decreases clinical score, histological score, and total alveoli area of mice mammary gland tissue (P < 0.01). (2) HMSeBA significantly increases TEER and fluorescein sodium leakage of HS-induced monolayer BMECs (P < 0.01), significantly improves the milk production and total area of alveoli (P < 0.01), and reduces clinical score, histological score, mRNA expression of heat stress-related proteins, and inflammatory cytokines release of heat-stressed mice (P < 0.01). (3) HMSeBA significantly improves tight junction structure damage, and significantly up-regulated the expression of tight junction proteins (ZO-1, claudin 1, and occludin) as well as signal molecules PI3K, AKT, and mTOR (P < 0.01) in heat-stressed mammary tissue. (4) HMSeBA significantly increases glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), and superoxide dismutase release (SOD) (P < 0.01) and significantly reduce malondialdehyde (MDA) expression (P < 0.01) in heat-stressed mammary tissue. In conclusion, this study implemented heat-stressed cell and mice model and showed that HMSeBA significantly regulate antioxidant capacity, inhibited inflammation, and regulate tight junction proteins expression in blood-milk barrier via PI3K/AKT/mTOR signaling pathway, so as to alleviate mammary gland damage and ensure its structure and function integrity.

Myosin Light Chain Kinase Modulates to Improve Myocardial Hypoxia/Reoxygenation Injury.

Objective The aim of this study was to evaluate whether myosin light chain kinase (MLCK) knockdown attenuated H9C2 cell hypoxia/reoxygenation (H/R) injury and downstream signaling pathway. Methods The MLCK expression in H/R injury model H9C2 cell was determined by western blot and qRT-PCR. H/R cells were transfected with si-MLCK in the presence of P38 inhibitor (SB203580) or ERK inhibitor (U0126). Then, cell apoptosis was verified by flow cytometry. Apoptosis-related proteins were detected by western blot. The contents of reactive oxygen species (ROS), lactate dehydrogenase (LDH), superoxide dismutase (SOD), interleukin-6 (IL-6), interleukin (IL)-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were measured using flow cytometry and colorimetric assays, respectively. Results MLCK expression was higher in H/R cells. Knockdown of MLCK diminished the amounts of ROS, LDH, IL-6, IL-1β, and TNF-α and elevated the release of SOD in H/R model H9C2 cells. Additionally, H/R injury induced the cumulative expression and phosphorylation of ERK and the phosphorylation of P38, whereas MLCK siRNA-treated cells showed decreased ERK1/2 and P38 activation. Inversely, P38 inhibitor (SB203580) and ERK inhibitor (U0126) could reverse the cardioprotective effects induced by si-MLCK. Conclusion MLCK knockdown attenuated H/R injury in H9C2 cells via regulating the ERK/P38 signaling pathway. MLCK/ERK/p38 axis may provide novel insight into therapeutic targets to restrain I/R injury caused by revascularization therapy after acute myocardial infarction.

Dioscin attenuates lipopolysaccharide-induced inflammatory myocardial injury through oxidative stress-related pathway

Lipopolysaccharide (LPS) is one of the main causes of myocardial injury. Dioscin has a protective effect on myocardial injury induced by LPS; however, the biological function and mechanism remain unclear. The purpose of this study was to investigate the effect of dioscin on myocardial injury induced by LPS. The myocardial injury model was constructed through LPS treatment of primary rat cardiomyocytes. Cardiomyocytes were treated with different concentrations of dioscin (50, 100, and 200 ng/mL). MTT was used to detect the activity of cardiomyocytes; flow cytometry and TUNEL assay were used to detect apoptosis; and enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). The release of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) was detected according to the kit instructions. The levels of apoptosis-related proteins (Bax, caspase-3, and Bcl 2) and the Nrf2-Keap1 pathway proteins were detected by western blot. Dioscin significantly reduced LPS-induced cardiomyocyte injury in neonatal rats in a concentration- and time-dependent manner. Dioscin also significantly inhibited cardiomyocyte inflammation and apoptosis induced by LPS. With the increase of dioscin concentration, reactive oxygen species (ROS) and MDA were downregulated, and SOD and GSH were upregulated. Moreover, dioscin inhibited LPS-induced myocardial injury by inhibiting the Nrf2-Keap1 pathway. Our study suggests that dioscin attenuates LPS-induced myocardial injury through oxidative stress-related pathways.

Fenton-like reaction performing mineralized nanocarriers as oxidative stress amplifying anticancer agents

We developed Fenton-like reaction performing mineralized nanoparticles as novel oxidative stress amplifying anticancer agents. We fabricated Cu-doped superoxide dismutase (SOD)-loaded calcium carbonate (CaCO3)-mineralized nanoparticles (Cu/SOD-MNPs) that could simultaneously release hydrogen peroxide-generating SOD and Fenton-like reaction-inducing copper ions within cancer cells. The CaCO3 core of the Cu/SOD-MNPs effectively inhibited the release of the entrapped SOD and copper ions in physiological pH (7.4). In contrast, in acidic pH (5.0), the release of SOD and copper ions was facilitated by CaCO3 dissolution. We demonstrated that the Cu/SOD-MNPs elevated the level of highly toxic apoptosis-inducing hydroxyl radicals in the cancer cells. The Cu/SOD-MNPs showed specific toxicity for cancer cells (MCF-7 cells), whereas no or negligible toxicity was found for normal cells (NIH3T3 and HEK293 cells). This mineralized oxidative stress amplifying nanoparticle may serve as a novel cancer-specific anticancer agent.

Comparison of Inflammatory Markers Between the Sevoflurane and Isoflurane Anesthesia in a Rat Model of Liver Ischemia/Reperfusion Injury

BackgroundSevoflurane and isoflurane had been reported to improve ischemia/reperfusion injury (I/R) through amelioration of the inflammatory response. We aimed to explore and compare the molecular mechanisms involved in sevoflurane and isoflurane anesthesia in liver ischemia-reperfusion of rat model. MethodsForty male Wistar rats were randomly divided into 4 groups: sham group, I/R group, sevoflurane group, and isoflurane group. The liver I/R injury model was established to investigate the effect of sevoflurane and isoflurane anesthesia on liver ischemia/reperfusion. The inflammatory markers and complement C3, C5a, and C6 were detected by enzyme-linked immunosorbent assay. Oxidative stress was detected by measuring the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO). ResultsOur results showed that sevoflurane anesthesia significantly decreased alanine transaminase, aspartate transaminase, and lactate dehydrogenase levels compared with isoflurane and controls. Sevoflurane inhibited I/R injury induced production of tumor necrosis factor α, interleukin 1, interleukin 6, and intercellular cell adhesion molecule-1 and promoted interleukin 10 production more significantly compared with isoflurane. Reduced MDA and NO and elevated SOD release suggested that oxidative stress was attenuated by sevoflurane and isoflurane anesthesia. Both sevoflurane and isoflurane anesthesia significantly decreased plasma C3 levels compared with the I/R injury group without differences. ConclusionSevoflurane anesthesia produced a more significant inhibitive effect on inflammatory cytokines and oxidative stress in liver I/R injury model than isoflurane, suggesting that sevoflurane is more suitable in surgery.

Bio-friendly encapsulation of superoxide dismutase into vaterite CaCO3 crystals. Enzyme activity, release mechanism, and perspectives for ophthalmology

Mesoporous vaterite CaCO3 crystals are nowadays one of the most popular vectors for loading of fragile biomolecules like proteins due to biocompatibility, high loading capacity, cost effective and simple loading procedures. However, recent studies reported the reduction of bioactivity for protein encapsulation into the crystals in water due to rather high alkaline pH of about 10.3 caused by the crystal hydrolysis. In this study we have investigated how to retain the bioactivity and control the release rate of the enzyme superoxide dismutase (SOD) loaded into the crystals via co-synthesis. SOD is widely used as an antioxidant in ophthalmology and its formulations with high protein content and activity as well as opportunities for a sustained release are highly desirable. Here we demonstrate that SOD co-synthesis can be done at pH 8.5 in a buffer without affecting crystal morphology. The synthesis in the buffer allows reaching the high loading efficiency of 93%, high SOD content (24 versus 15 w/w % for the synthesis in water), and order of magnitude higher activity compared to the synthesis in water. The enormous SOD concentration into crystals of 10−2 M is caused by the entrapment of SOD aggregates into the crystal pores. The SOD released from crystals at physiologically relevant ionic strength fully retains its bioactivity. As found by fitting the release profiles using zero-order and Baker-Lonsdale models, the SOD release mechanism is governed by both the SOD aggregate dissolution and by the diffusion of SOD molecules thorough the crystal pores. The latest process contributes more in case of the co-synthesis in the buffer because at higher pH (co-synthesis in water) the unfolded SOD molecules aggregate stronger. The release is bi-modal with a burst (ca 30%) followed by a sustained release and a complete release due to the recrystallization of vaterite crystals to non-porous calcite crystals. The mechanism of SOD loading into and release from the crystals as well as perspectives for the use of the crystals for SOD delivery in ophthalmology are discussed. We believe that together with a fundamental understanding of the vaterite-based protein encapsulation and protein release, this study will help to establish a power platform for a mild and effective encapsulation of fragile biomolecules like proteins at bio-friendly conditions.

Hydrogen‑rich solution against myocardial injury and aquaporin expression via the PI3K/Akt signaling pathway during cardiopulmonary bypass in rats.

Myocardial ischemia, hypoxia and reperfusion injury are induced by aortic occlusion, cardiac arrest and resuscitation during cardiopulmonary bypass (CPB), which can severely affect cardiac function. The aim of the present study was to investigate the effects of hydrogen-rich solution (HRS) and aquaporin (AQP) on cardiopulmonary bypass (CPB)-induced myocardial injury, and determine the mechanism of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Sprague Dawley rats were divided into a sham operation group, a CPB surgery group and a HRS group. A CPB model was established, and the hemodynamic parameters were determined at the termination of CPB. The myocardial tissues were observed by hematoxylin and eosin, and Masson staining. The levels of myocardial injury markers [adult cardiac troponin I (cTnI), lactate dehydrogenase (LDH), creatine kinase MB (CK-MB) and brain natriuretic peptide (BNP)], inflammatory factors [interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α)] and oxidative stress indicators [superoxide dismutase (SOD), malondialdehyde (MDA) and myeloperoxidase (MPO)] were determined by ELISA. Furthermore, H9C2 cells were treated with HRS following hypoxia/reoxygenation. Cell viability and cell apoptosis were investigated. The expression of apoptosis regulator Bcl-2 (Bcl-2), apoptosis regulator Bax (Bax), caspase 3, AQP-1, AQP-4, phosphorylated (p)-Akt, heme oxygenase 1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) were investigated using western blotting and quantitative-polymerase chain reaction of tissues and cells. Following CPB, myocardial cell arrangement was disordered, myocardial injury markers (cTnI, LDH, CK-MB and BNP), inflammatory cytokines (IL-1β, IL-6 and TNF-α) and MDA levels were significantly increased compared with the sham group; whereas the SOD levels were significantly downregulated following CPB compared with the sham group. HRS attenuated myocardial injury, reduced the expression levels of cTnI, LDH, CK-MB, BNP, IL-1β, IL-6, TNF-α, MDA and MPO, and increased SOD release. Levels of Bcl-2, AQP-1, AQP-4, p-Akt, HO-1 and Nrf2 were significantly increased following HRS; whereas Bax and caspase-3 expression levels were significantly reduced following CPB. HRS treatment significantly increased the viability of myocardial cells, reduced the rate of myocardial cell apoptosis and the release of MDA and LDH compared with the CPB group. A PI3K inhibitor (LY294002) was revealed to reverse the protective effect of HRS treatment. HRS was demonstrated to attenuate CPB-induced myocardial injury, suppress AQP-1 and AQP-4 expression following CPB treatment and protect myocardial cells via the PI3K/Akt signaling pathway.

The Effects of Two Anesthetics, Propofol and Sevoflurane, on Liver Ischemia/Reperfusion Injury

Background: Propofol and sevoflurane are widely used in clinical anesthesia, and both have been reported to exert a protective effect in organ ischemia/reperfusion (IR). This study aims to investigate and compare the effects of propofol and sevoflurane on liver ischemia/reperfusion and the precise molecular mechanism. Methods and Materials: Rats were randomized into four groups: the sham group, I/R group, propofol treatment group (infused with 1% propofol at 500 µg· kg-1· min-1), and sevoflurane treatment group (infused with 3% (2 L/min) sevoflurane). The liver ischemia/reperfusion model was used to evaluate the hepatoprotective effect on ischemic injury. Liver enzyme leakage, liver cytokines and histopathological examination were used to evaluate the extent of hepatic ischemia/reperfusion injury. Oxidative stress was investigated by evaluating the levels of Malondialdehyde(MDA), Superoxide Dismutase(SOD) and NO. The terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling (TUNEL) assay and western blot were applied to detect apoptosis in the ischemic liver tissue and its mechanism. Results: Both propofol and sevoflurane attenuated the extent of hepatic ischemia/reperfusion injury which is evident from the hisopathological studies and alterations in liver enzymes such as AST and LDH by inhibiting Nuclear factor kappa B (NFκB) activation and subsequent alterations in inflammatory cytokines interleukin-1(IL-1), interleukin-6(IL-6), tumor necrosis factor-alpha (TNF-a) and increased IL10 release. Propofol exhibited a similar protective effect and a lower IL-1 release, while sevoflurane decreased TNF-a leakage more significantly. Meanwhile, oxidative stress was attenuated by reduced MDA and NO and elevated SOD release. The expression of antiapoptotic protein Bcl-2 and Bcl-xl were enhanced while that of apoptotic protein Bax and Bak were reduced by both propofol and sevoflurane to regulate hepatic apoptosis. In addition, propofol downregulated the phosphorylation of AKT and Bad protein, while sevoflurane downregulated the phosphorylation of p38. In addition, Both the treatments had no effect on the expression of AKT, Bad and p38. Conclusion: Both propofol and sevoflurane can protect the liver from ischemia/reperfusion injury by modulating the inflammatory responses reducing oxidative stress and liver apoptosis.

A poly(γ‐glutamic acid)‐based hydrogel loaded with superoxide dismutase for wound healing

ABSTRACTThe purpose of this study was to develop a poly(γ‐glutamic acid) (γ‐PGA)‐based hydrogel loaded with superoxide dismutase (SOD) to accelerate wound healing. First, γ‐PGA was modified with taurine (γ‐PGAS), and then the SOD‐loaded γ‐PGAS/γ‐PGA hydrogel (SOD‐PGAS/PGA‐H) was prepared by cross‐linking of ethylene glycol diglycidyl ether. The swelling behavior and water vapor transmission rate revealed that PGAS/PGA‐H could create a moist environment for wound surface.In vitrokinetics of SOD release showed that SOD released from PGAS/PGA‐H maintained high activity and SOD‐PGAS/PGA‐H effectively scavenged the superoxide anion. The results of our fibroblast proliferation experiments showed that PGAS/PGA‐H had good cytocompatibility. The effects of SOD‐PGAS/PGA‐H on wound healing were examined in a Type I diabetic rat model with full‐thickness wounds. Twenty‐one days after grafted to wounds, SOD‐PGAS/PGA‐H exhibited a higher rate of wound healing than control group and showed increased collagen deposition and epithelialization. SOD‐PGAS/PGA‐H seems to promote better wound healing and thus might be a promising candidate for wound healing management. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci.2015,132, 42033.

Protective role of adiponectin in a rat model of intestinal ischemia reperfusion injury.

To determine the potential protective role of adiponectin in intestinal ischemia reperfusion (I/R) injury. A rat model of intestinal I/R injury was established. The serum level of adiponectin in rats with intestinal I/R injury was determined by enzyme-linked immunosorbent assay (ELISA). The serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were also measured by ELISA. Apoptosis of intestinal cells was detected using the terminal deoxynucleotidyl transferase dUTP nick end labeling assay. The production of malondialdehyde (MDA) and superoxide dismutase (SOD) and villous injury scores were also measured. Adiponectin was downregulated in the serum of rats with intestinal I/R injury compared with sham rats. No significant changes in the expression of adiponectin receptor 1 and adiponectin receptor 2 were found between sham and I/R rats. Pre-treatment with recombinant adiponectin attenuated intestinal I/R injury. The production of pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α, in rats with intestinal I/R injury was reduced by adiponectin pre-treatment. The production of MDA was inhibited, and the release of SOD was restored by adiponectin pre-treatment in rats with intestinal I/R injury. Adiponectin pre-treatment also inhibited cell apoptosis in these rats. Treatment with the AMP-activated protein kinase (AMPK) signaling pathway inhibitor, compound C, or the heme oxygenase 1 (HO-1) inhibitor, Snpp, attenuated the protective effects of adiponectin against intestinal I/R injury. Adiponectin exhibits protective effects against intestinal I/R injury, which may involve the AMPK/HO-1 pathway.

Effect of Nonenzymatic Antioxidants on Sperm Motility and Survival Relative to Free Radicals and Antioxidant Enzymes of Chilled-Stored Ram Semen

The effect of inclusion of three antioxidants (Vitamin E, cysteine and glutathione) in ram semen extender on the release of antioxidant enzymes and free radicals was studied. A 3 × 3 factorial experiment was conducted to test effects of supplementing ram semen extender with Vitamin E (1, 5 and 10 IU), cysteine (1, 5 and 10 mM) or glutathione (0.5, 1 and 2 mM) on the sperm survival and release of superoxide dismutase (SOD), glutathione peroxidase (GPX) and thiobarbituric acid reactive substances (TBARS). Eighty ejaculates of eight fertile Najdi rams were collected, assessed for the gross examination and the good ejaculates (≥90% motility) were pooled and sperm count was assessed. Therefore ten extenders; control (C) and 9 treated (C plus antioxidant) were tested. Extended semen was stored at 5°C for 96 h, examined for motility and survival and sperm cells were separated from plasma, sonicated, homogenized and exposed to the determinations of SOD, GPX and TBARS. The highest sperm survival was found in diluents containing 5 IU Vitamin E/ml, 1 and 2 mM glutathione (55.5% survival), while the lowest survival was found in 10 mM cysteine (11.1%). TBARS concentration was highest (P < 0.05) in control than other treatments, however, Vitamin E and glutathione exhibited low values. Contrariwise, activities of SOD and GPX increased (P < 0.05) within sperm cells and seminal plasma in diluents containing 5 IU E, 1 mM and 2 mM glutathione. The enzymatic activities were generally higher in seminal plasma than in sperm cells. It was concluded that supplementing ram semen extender during chilled storage with 5 IU Vitamin E per ml or 1 - 2 mM glutathione enhanced sperm survival and reduced free radicals.

Superoxide Dismutase Binding and Release Behaviors of Dodecylated Poly(allylamine)s: Effects of Self-Aggregation and Organic Solvents

pH sensitive dodecyl-modified poly(allylamine)s associate with superoxide dismutase (SOD) in aqueous solutions at pH 5.5 and 7.4. The bound SOD can be released completely at pH 10.0 due to the self-aggregation of the polymers. In a mixture of water and organic solvent, the hydrophobic groups of the polymers are exposed. During the process of removing the organic solvent, the polymers increase their hydrophobic interactions with SOD. As a result, the SOD binding of the polymers increases; the release of SOD from the complexes is remarkably extended. The released SOD molecules have full activity. The knowledge gained in this study will be beneficial to the applications of hydrophobically modified polyelectrolytes as delivery carriers for protein/peptide drugs.

Learning from your mistakes: is aneuploidy so bad, after all?

Learning from your mistakes: is aneuploidy so bad, after all?

Bioactive Long-Term Release from Biodegradable Microspheres Preserves Implanted ALG-PLO-ALG Microcapsules from In Vivo Response to Purified Alginate

To assess whether prevention of unexpected in vivo adverse inflammatory and immune responses to biohybrid organ grafts for the treatment of Type I Diabetes Mellitus (T1DM) is possible by superoxide dismutase and ketoprofen controlled release. Superoxide dismutase and ketoprofen-loaded polyester microspheres were prepared by W/O/W and O/W methods, embodied into purified alginate-poly-L-ornithine-alginate microcapsules and intraperitoneally implanted into CD1 mice. The microspheres were characterized for morphology, size, encapsulation efficiency, enzyme activity and in vitro release. Purified alginate contaminants were assayed, and the obtained microcapsules were investigated for size and morphology before and after implantation over 30 days. Cell pericapsular overgrowth and expression were evaluated by optical microscopy and flow cytometry. Superoxide dismutase and ketoprofen sustained release reduced cell pericapsular overgrowth in comparison to the control. Superoxide dismutase release allowed preserving the microcapsules over 30 days. Ketoprofen-loaded microspheres showed some effect in the immediate post-grafting period. A higher macrophage and T-cell expression was observed for the control group. Microspheres containing superoxide dismutase and ketoprofen may represent novel tools to limit or prevent unpredictable adverse in vivo response to alginate, thus contributing to improve cell transplantation success rates in T1DM treatment.

Sodium salicylate switches glucose depletion-induced necrosis to autophagy and inhibits high mobility group box protein 1 release in A549 lung adenocarcinoma cells

Sodium salicylate, the active metabolite of aspirin, has been shown to exert anti-inflammatory activities by inhibiting the expression of various pro-inflammatory factors, and has potent anti-cancer effects against a number of human cancers including colon, lung, breast and leukemia. Necrotic cell death is emerging as one of the crucial factors that trigger an inflammatory response since during necrotic death the cell membrane is ruptured and the intracellular constituents including high mobility group box 1 (HMGB1) are released into the extracellular space, thereby activating an inflammatory response. In contrast, autophagic death is regarded as a form of tumour suppressive cell death, as indicated in tumour suppressors such as beclin 1 in autophagic pathways. To better understand the anti-inflammatory properties of sodium salicylate and its effect on necrotic cell death in A549 cells induced by glucose depletion (GD), a common characteristic of the tumour micro-environment, was examined. While GD induced mostly necrotic death in A549 cells, salicylate suppresssed GD-induced necrosis and HMGB1 release. In addition, salicylate shifted the cell death pattern to autophagy by inhibiting GD-induced Cu/Zn superoxide dismutase release and ROS production. These results indicate that the activity of salicylate to prevent necrotic death may contribute to its anti-inflammatory action and suppress tumour development possibly through switching the cell death mode from tumour-promoting necrotic cell death to tumour-suppressive autophagic cell death.

Assessment of physical and antioxidant activity stability, in vitro release and in vivo efficacy of formulations added with superoxide dismutase alone or in association with α-tocopherol

A topical formulation was added with different concentrations of superoxide dismutase (SOD) alone or in association with α-tocopherol (α-TOC). The physical stability was evaluated by rheological behavior of formulations stored at 4 °C, 30 °C/60% RH and 40 °C/70% RH for 6 months. SOD alone and formulations containing SOD 0.2%, 0.4% or 0.6% or SOD and α-TOC were stored in the same conditions and the enzymatic activity was evaluated by the superoxide anion scavenging using chemiluminescence measurement. In vitro release study was carried out using modified Franz diffusion cell and SOD formulations photoprotection against skin erythema was observed for 72 h. SOD and α-TOC formulation proved to be instable, since the interaction between the antioxidants led to both physical and enzymatic activity instability. SOD formulations showed to be physically stable and maintained the enzymatic activity for 6 months when stored at 4 and 30 °C/60% RH. Despite the fact of low SOD release from the formulation, it was effective in inhibiting the UVB-induced skin erythema for 48 h after a single application. Topical administration of antioxidants provides an efficient way to enrich the endogenous cutaneous protection system, and SOD formulations could be used for improving photoprotection of skin.

Microencapsulation of Superoxide Dismutase into Poly(ε-Caprolactone) Microparticles by Reverse Micelle Solvent Evaporation

The aim of this work was to encapsulate superoxide dismutase (SOD) in poly(ε-caprolactone) (PCL) microparticles by reverse micelle solvent evaporation. The concentration of PCL, the hydrophile-lipophile balance (HLB), and concentration of the sucrose ester used as surfactant in the organic phase were investigated as formulation variables. Relatively higher encapsulation efficiency (∼48%) and retained enzymatic activity (>90%) were obtained with microparticle formulation made from the 20% (w/v) PCL and 0.05% (w/v) sucrose ester of HLB = 6. This formulation allowed the in vitro release of SOD for at least 72 hr. These results showed that reverse micelle solvent evaporation can be used to efficiently encapsulate SOD in PCL microparticles. Such formulations may improve the bioavailability of SOD.