An Novel Approach in Protective Gene Therapy: Recombinant Adeno-Associated Virus 2-Mediated Transfer of the Human Superoxide-Dismutase Gene.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 5265-5265
Author(s):  
Marlon R. Veldwijk ◽  
Carsten Herskind ◽  
Stephanie Laufs ◽  
Marius Stiefelhagen ◽  
W. Jens Zeller ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Therapy ◽  
Superoxide Dismutase ◽  
Tumor Cells ◽  
Normal Tissue ◽  
Therapeutic Window ◽  
Cervix Carcinoma ◽  
Sod Activity ◽  
Adeno Associated Virus ◽  
Tissue Protection

Abstract Background and purpose: The success rate of any therapeutic approach depends on the therapeutic window, which can be increased by either raising the resistance of the normal tissue such as hematopoietic cells without protecting the tumor cells or by sensitizing the tumor cells but not the normal cells. Two promising candidate genes for normal tissue protection against superoxide-induced damage may be the copper-zinc (CuZnSOD) and manganese superoxide-dismutase genes (MnSOD). Oxidative stress-induced apoptosis plays a role in both radiation- and chemotherapy-induced tissue damage. Relevant cytostatic drugs for which oxidative stress as one mechanism of action has been described are e.g. anthracyclins, platinum analogues, etoposide and ara-C. For gene therapeutic approaches, recombinant adeno-associated virus 2 (rAAV-2)-based vectors offer attractive advantages over other vector systems: low immunogenicity, possibility of in vivo application, ability to infect dividing and non-dividing tissues and a low chance of insertional mutagenesis, due to extra-chromosomal localization. Here, we report the production and testing of novel rAAV-2-SOD vectors with the goal of normal tissue protection. Material and methods: Various rAAV-2 vectors containing CuZnSOD, MnSOD and fusion proteins of both with the enhanced green fluorescent protein (eGFP) gene were cloned and vector stocks were produced. Human cervix carcinoma (HeLa-RC) cells were chosen for their susceptibility to rAAV-2. Cells were seeded and transduced with the rAAV-2-SOD vectors. Gene transfer and transgene expression were investigated using FACS and an SOD-activity assay. Results: Over 70% of all HeLa cells expressed SOD and significant amounts of functional SOD protein were detected (table 1). Conclusion: These results forms the basis to evaluate the radio- and chemoprotective effects of AAV-mediated SOD gene therapy in hematopoietic and non-hematopoietic (e.g. mucosal) cells. Vector % GFP+ cells SOD activity (U/mg) Mock control - 435 ± 37 rAAV-2-CuZnSOD N/A 965 ± 112 rAAV-2-CuZnSOD/eGFP 78 ± 2 1093 ± 178 rAAV-2-MnSOD N/A 1516 ± 191 rAAV-2-MnSOD/eGFP 77 ± 3 1204 ± 124

10-gingerol induces oxidative stress through HTR1A in cumulus cells: in-vitro and in-silico studies

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kiptiyah Kiptiyah ◽  
Widodo Widodo ◽  
Gatot Ciptadi ◽  
Aulanni’am Aulanni’Am ◽  
Mohammad A. Widodo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Culture ◽  
Superoxide Dismutase ◽  
In Silico ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Sod Activity ◽  
In Silico Studies ◽  
Incubation Periods

AbstractBackgroundWe investigated whether 10-gingerol is able to induce oxidative stress in cumulus cells.MethodsFor the in-vitro research, we used a cumulus cell culture in M199, containing 10-gingerol in various concentrations (0, 12, 16, and 20 µM), and detected oxidative stress through superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations, with incubation periods of 24, 48, 72, and 96 h. The obtained results were confirmed by in-silico studies.ResultsThe in-vitro data revealed that SOD activity and MDA concentration increased with increasing incubation periods: SOD activity at 0 µM (1.39 ± 0.24i), 12 µM (16.42 ± 0.35ab), 16 µM (17.28 ± 0.55ab), 20 µM (17.81 ± 0.12a), with a contribution of 71.1%. MDA concentration at 0 µM (17.82 ± 1.39 l), 12 µM (72.99 ± 0.31c), 16 µM (79.77 ± 4.19b), 20 µM (85.07 ± 2.57a), with a contribution of 73.1%. Based on this, the in-silico data uncovered that 10˗gingerol induces oxidative stress in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.Conclusions10-gingerol induces oxidative stress in cumulus cells through enhancing SOD activity and MDA concentration by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.

Protective Effects of Ginsenosides on 17α-Ethynyelstradiol-Induced Intrahepatic Cholestasis via Anti-Oxidative and Anti-Inflammatory Mechanisms in Rats

2017 ◽  
Vol 45 (08) ◽  
pp. 1613-1629 ◽  
Author(s):  
Yan-Jiao Xu ◽  
Zao-Qin Yu ◽  
Cheng-Liang Zhang ◽  
Xi-Ping Li ◽  
Cheng-Yang Feng ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Alkaline Phosphatase ◽  
Superoxide Dismutase ◽  
Protein Expression ◽  
Intrahepatic Cholestasis ◽  
Serum Levels ◽  
Total Bile Acid ◽  
Protective Effects ◽  
Sod Activity ◽  
Mda Content

The present study was designed to assess the effects and potential mechanisms of ginsenosides on 17[Formula: see text]-ethynyelstradiol (EE)-induced intrahepatic cholestasis (IC). Ginsenoside at doses of 30, 100, 300[Formula: see text]mg/kg body weight was intragastrically (i.g.) given to rats for 5 days to examine the effect on EE-induced IC. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bile acid (TBA) were measured. Hepatic malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. Protein expression of proinflammatory cytokines TNF-[Formula: see text], IL-6 and IL-1[Formula: see text] was analyzed by immunohistochemistry and Western blot. Results indicated that ginsenosides remarkably prevented EE-induced increase in the serum levels of AST, ALT, ALP and TBA. Moreover, the elevation of hepatic MDA content induced by EE was significantly reduced, while hepatic SOD activities were significantly increased when treated with ginsenosides. Histopathology of the liver tissue showed that pathological injuries were relieved after treatment with ginsenosides. In addition, treatment with ginsenosides could significantly downregulate the protein expression of TNF-[Formula: see text], IL-6 and IL-1[Formula: see text] compared with EE group. These findings indicate that ginsenosides exert the hepatoprotective effect on EE-induced intrahepatic cholestasis in rats, and this protection might be attributed to the attenuation of oxidative stress and inflammation.

The state of the blood antioxidant system in the patients presenting with acromegaly

2015 ◽  
Vol 61 (2) ◽  
pp. 8-11
Author(s):  
M V Faassen ◽  
M S Pankratova ◽  
N N Molitvoslovova ◽  
A A Baizhumanov ◽  
S S Kovalenko ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Activity ◽  
Superoxide Dismutase ◽  
Blood Plasma ◽  
Antioxidant System ◽  
The State ◽  
Total Antioxidant Activity ◽  
Sod Activity ◽  
Protein Thiols ◽  
Total Antioxidant

The state of the blood antioxidant system in the patients presenting with acromegaly. The objective of the present study was to evaluate the total antioxidant activity (TAA) of blood plasma, the levels of non-protein thiols and 2-thyobarbituric acid-active products (TBA-AP), superoxide dismutase (SOD) and catalase (CAT) activities as well as ceruloplasmin (CP) level in the patients presenting with acromegaly. It was shown that plasma TAA and SOD activity in this patients was on the average 20 and 30% lower respectively than in the control subjects. At the same time, the TBA-AP and CP levels increased by 50 and 40% respectively. These data suggest the development of oxidative stress in the acromegalic patients.

Further investigation of the mechanism of Vitreoscilla hemoglobin (VHb) protection from oxidative stress in Escherichia coli

Biologia ◽  
2011 ◽  
Vol 66 (5) ◽  
Author(s):  
Meltem Akbas ◽  
Tugrul Doruk ◽  
Serhat Ozdemir ◽  
Benjamin Stark
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Superoxide Dismutase ◽  
Stationary Phase ◽  
Exponential Phase ◽  
Vitreoscilla Hemoglobin ◽  
Sod Activity ◽  
E Coli ◽  
Hydrogen Peroxide Treatment

AbstractIn Escherichia coli, Vitreoscilla hemoglobin (VHb) protects against oxidative stress, perhaps, in part, by oxidizing OxyR. Here this protection, specifically VHb-associated effects on superoxide dismutase (SOD) and catalase levels, was examined. Exponential or stationary phase cultures of SOD+ or SOD− E. coli strains with or without VHb and oxyR antisense were treated with 2 mM hydrogen peroxide without sublethal peroxide induction, and compared to untreated control cultures. The hydrogen peroxide treatment was toxic to both SOD+ and SOD− cells, but much more to SOD− cells; expression of VHb in SOD+ strains enhanced this toxicity. In contrast, the presence of VHb was generally associated in the SOD+ background with a modest increase in SOD activity that was not greatly affected by oxyR antisense or peroxide treatment. In both SOD+ and SOD− backgrounds, VHb was associated with higher catalase activity both in the presence and absence of peroxide. Contrary to its stimulatory effects in stationary phase, in exponential phase oxyR antisense generally decreased VHb levels.

scholarly journals Effect of Nandrolone Treatment with And Without Resistance Training on Superoxide Dismutase Concentration and Pathology of Kidney Tissue in Rats

2020 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Esfandiyar Heidari ◽  
Seyed Ali Hosseini ◽  
Mohammad Ali Azarbayjani
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Resistance Training ◽  
Anabolic Steroids ◽  
Kidney Tissue ◽  
Sod Activity ◽  
Significant Difference ◽  
Stress Levels ◽  
Sh Group ◽  
Male Wistar Rats

Background: The prevalence of anabolic steroids abuse in athletes and non-athletes is associated with the risk of injury to various organs, but there are limited studies of oxidative changes in kidney tissue following nandrolone (N) administration. Objectives: the aim of this study was to investigate the effect of N treatment with and without resistance training (RT) on superoxide dismutase (SOD) concentration and tissue pathology of kidney tissue in rats. Methods: In this experimental study, 20 male Wistar rats were randomly divided into four groups of five rats including 1) control (C), 2) sham (normal saline) (Sh), 3) N, and 4) N + RT. Groups 3 and 4 received 10 mg/kg N peritoneally, and the N + RT group performed 1 m ladder climbing for eight weeks and three sessions per week. SOD levels of kidney tissue were measured by ELISA and radioimmunoassay. Hematoxylin-eosin (H&E) staining was used to evaluate oxidative stress levels in kidney tissue. One-way ANOVA with Bonferroni’s post- hoc tests were used for analysis of research findings in SPSS version 22 (P ≤ 0.05). Results: SOD levels in the C group were higher than the Sh (P = 0.001), N (P = 0.001), and N + RT (P = 0.001) groups. SOD levels were lower in the Sh group than in the N (P = 0.049) and N + RT (P = 0.001) groups. However, there was no significant difference in SOD levels in the N + RT group and N group (P = 0.28). Also, oxidative stress levels were normal in tissue studies in all groups. Conclusions: It seems that Ntreatment with and without RT reduces SOD activity in kidney tissue, but more studies are needed in this regard given the normality of tissue oxidative stress results.

scholarly journals Natural Compounds Rosmarinic Acid and Carvacrol Counteract Aluminium-Induced Oxidative Stress

Molecules ◽  
2020 ◽  
Vol 25 (8) ◽  
pp. 1807 ◽  
Author(s):  
Juste Baranauskaite ◽  
Ilona Sadauskiene ◽  
Arunas Liekis ◽  
Arturas Kasauskas ◽  
Robertas Lazauskas ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Superoxide Dismutase ◽  
Phenolic Compounds ◽  
Rosmarinic Acid ◽  
Sod Activity ◽  
Mice Brain ◽  
Aluminum Accumulation ◽  
Liver Homogenates ◽  
The Brain

Aluminum accumulation, glutathione (GSH) and malondialdehyde (MDA) concentrations as well as catalase (CAT) and superoxide dismutase (SOD) activities were determined in erythrocytes and brain and liver homogenates of BALB/c mice treated with Al3+ (7.5 mg/kg/day (0.15 LD50) as AlCl3 (37.08 mg/kg/day), whereas HCl (30.41 mg/kg/day) was used as Cl− control, the treatments were performed for 21 days, i.p., in the presence and absence of rosmarinic acid (0.2805 mg/kg/day (0.05 LD50), 21 days, i.g.) or carvacrol (0.0405 mg/kg/day (0.05 LD50), 21 days, i.g.). The treatment with AlCl3 increased GSH concentration in erythrocytes only slightly and had no effect on brain and liver homogenates. Rosmarinic acid and carvacrol strongly increased GSH concentration in erythrocytes but decreased it in brain and liver homogenates. However, AlCl3 treatment led to Al accumulation in mice blood, brain, and liver and induced oxidative stress, assessed based on MDA concentration in the brain and liver. Both rosmarinic acid and carvacrol were able to counteract the negative Al effect by decreasing its accumulation and protecting tissues from lipid peroxidation. AlCl3 treatment increased CAT activity in mice brain and liver homogenates, whereas the administration of either rosmarinic acid or carvacrol alone or in combination with AlCl3 had no significant effect on CAT activity. SOD activity remained unchanged after all the treatments in our study. We propose that natural herbal phenolic compounds rosmarinic acid and carvacrol could be used to protect brain and liver against aluminum induced oxidative stress leading to lipid peroxidation.

scholarly journals Dual AAV-mediated gene therapy restores hearing in a DFNB9 mouse model

2019 ◽  
Vol 116 (10) ◽  
pp. 4496-4501 ◽  
Author(s):  
Omar Akil ◽  
Frank Dyka ◽  
Charlotte Calvet ◽  
Alice Emptoz ◽  
Ghizlene Lahlou ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Mouse Model ◽  
Autosomal Recessive ◽  
Therapeutic Option ◽  
The Other ◽  
Therapeutic Window ◽  
Congenital Deafness ◽  
Adeno Associated Virus ◽  
Coding Sequence ◽  
Packaging Capacity

Autosomal recessive genetic forms (DFNB) account for most cases of profound congenital deafness. Adeno-associated virus (AAV)-based gene therapy is a promising therapeutic option, but is limited by a potentially short therapeutic window and the constrained packaging capacity of the vector. We focus here on the otoferlin gene underlying DFNB9, one of the most frequent genetic forms of congenital deafness. We adopted a dual AAV approach using two different recombinant vectors, one containing the 5′ and the other the 3′ portions of otoferlin cDNA, which exceed the packaging capacity of the AAV when combined. A single delivery of the vector pair into the mature cochlea ofOtof−/−mutant mice reconstituted the otoferlin cDNA coding sequence through recombination of the 5′ and 3′ cDNAs, leading to the durable restoration of otoferlin expression in transduced cells and a reversal of the deafness phenotype, raising hopes for future gene therapy trials in DFNB9 patients.

Superoxide dismutase mimetic drug tempol aggravates anti-GBM antibody-induced glomerulonephritis in mice

2010 ◽  
Vol 299 (2) ◽  
pp. F445-F452 ◽  
Author(s):  
Hua Lu ◽  
Junhui Zhen ◽  
Tianfu Wu ◽  
Ai Peng ◽  
Ting Ye ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Superoxide Dismutase ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Immunohistochemical Staining ◽  
Treated Group ◽  
First Line ◽  
Sod Activity ◽  
Leukocyte Influx

Oxidative stress plays an important role in the pathogenesis of anti-glomerular basement membrane antibody-induced glomerulonephritis (anti-GBM-GN). Superoxide dismutase (SOD) is the first line of defense against oxidative stress by converting superoxide to hydrogen peroxide (H2O2). We investigated the effect of the SOD mimetic drug tempol on anti-GBM-GN in mice. 129/svJ mice were challenged with rabbit anti-mouse-GBM sera to induce GN and subsequently divided into tempol (200 mg·kg−1·day−1, orally) and vehicle-treated groups. Routine histology, SOD and catalase activities, malondialdehyde (MDA), H2O2, and immunohistochemical staining for neutrophils, lymphocytes, macrophages, p65-NF-κB, and osteopontin were performed. Mice with anti-GBM-GN had significantly reduced renal SOD and catalase activities and increased H2O2 and MDA levels. Unexpectedly, tempol administration exacerbated anti-GBM-GN as evidenced by intensification of proteinuria, the presence of severe crescentic GN with leukocyte influx, and accelerated mortality in the treated group. Tempol treatment raised SOD activity and H2O2 level in urine, upregulated p65-NF-κB and osteopontin in the kidney, but had no effect on renal catalase activity. Thus tempol aggravates anti-GBM-GN by increasing production of H2O2 which is a potent NF-κB activator and as such can intensify inflammation and renal injury. This supposition is supported by increases seen in p65-NF-κB, osteopontin, and leukocyte influx in the kidneys of the tempol-treated group.

scholarly journals Glutathione Peroxidase (GPx) and Superoxide Dismutase (SOD) in Oropharyngeal Cancer Associated with EBV and HPV Coinfection

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 1008
Author(s):  
Małgorzata Strycharz-Dudziak ◽  
Sylwia Fołtyn ◽  
Jakub Dworzański ◽  
Małgorzata Kiełczykowska ◽  
Maria Malm ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Glutathione Peroxidase ◽  
Oropharyngeal Cancer ◽  
Hpv Infection ◽  
Cancer Tissue ◽  
Upper Aerodigestive Tract ◽  
Sod Activity ◽  
Tissue Samples ◽  
Hpv Dna

Recent reports have pointed to the link between persistent inflammation, oxidative stress, and carcinogenesis; however most of the studies concerning the role of viruses in head and neck cancer (HNC) are focused mainly on one type of virus. Our present study aimed to study the relationship between Epstein–Barr virus/human papilloma virus (EBV/HPV) coinfection and glutathione peroxidase (GPx) and superoxide dismutase (SOD) level in oropharyngeal cancer. Fresh-frozen tumor tissue samples were collected from 128 patients with oropharyngeal cancer infected with EBV or HPV or with EBV/HPV coinfection. After DNA extraction, EBV and HPV DNA was detected using a polymerase chain reaction (PCR) assay. GPx and SOD activity was determined in homogenates of cancer tissue using diagnostic kits produced by Randox Laboratories. Both GPx and SOD activity was statistically lower in patients with EBV/HPV coinfection than in a single EBV or HPV infection. Analysis of GPx and SOD activity in relation to histological grading and tumor, node (TN) classification revealed that in poorly-differentiated tumors, the level of antioxidant enzymes was lower compared with well-differentiated lesions and in cases with greater tumor dimensions and lymph-node involvement, both GPx and SOD activity was decreased. Further studies are necessary to clarify the influence of interplay between EBV, HPV, and oxidative stress on malignant transformation of upper aerodigestive tract epithelial cells.

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