SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult

Succinate semialdehyde dehydrogenase (SSADH) is a mitochondrial enzyme, encoded by ALDH5A1, mainly involved in γ-aminobutyric acid (GABA) catabolism and energy supply of neuronal cells, possibly contributing to antioxidant defense. This study aimed to further investigate the antioxidant role of SSADH, and to verify if common SNPs of ALDH5A1 may affect SSADH activity, stability, and mitochondrial function. In this study, we used U87 glioblastoma cells as they represent a glial cell line. These cells were transiently transfected with a cDNA construct simultaneously harboring three SNPs encoding for a triple mutant (TM) SSADH protein (p.G36R/p.H180Y/p.P182L) or with wild type (WT) cDNA. SSADH activity and protein level were measured. Cell viability, lipid peroxidation, mitochondrial morphology, membrane potential (ΔΨ), and protein markers of mitochondrial stress were evaluated upon Paraquat treatment, in TM and WT transfected cells. TM transfected cells show lower SSADH protein content and activity, fragmented mitochondria, higher levels of peroxidized lipids, and altered ΔΨ than WT transfected cells. Upon Paraquat treatment, TM cells show higher cell death, lipid peroxidation, 4-HNE protein adducts, and lower ΔΨ, than WT transfected cells. These results reinforce the hypothesis that SSADH contributes to cellular antioxidant defense; furthermore, common SNPs may produce unstable, less active SSADH, which could per se negatively affect mitochondrial function and, under oxidative stress conditions, fail to protect mitochondria.

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Pathway of γ-Aminobutyrate Metabolism in Rhizobium leguminosarum 3841 and Its Role in Symbiosis

Journal of Bacteriology ◽

10.1128/jb.01714-08 ◽

2009 ◽

Vol 191 (7) ◽

pp. 2177-2186 ◽

Cited By ~ 39

Author(s):

Jurgen Prell ◽

Alexandre Bourdès ◽

Ramakrishnan Karunakaran ◽

Miguel Lopez-Gomez ◽

Philip Poole

Keyword(s):

Nitrogen Source ◽

Rhizobium Leguminosarum ◽

Tricarboxylic Acid Cycle ◽

Amino Group ◽

Triple Mutant ◽

Gaba Aminotransferase ◽

Semialdehyde Dehydrogenase ◽

Elevated Expression ◽

Succinate Semialdehyde ◽

Cultured Bacteria

ABSTRACT Pea plants incubated in 15N2 rapidly accumulated labeled γ-aminobutyrate (GABA) in the plant cytosol and in bacteroids of Rhizobium leguminosarum bv. viciae 3841. Two pathways of GABA metabolism were identified in R. leguminosarum 3841. In the first, glutamate is formed by GABA aminotransferase (GabT), transferring the amino group from GABA to 2-oxoglutarate. In the second, alanine is formed by two ω-aminotransferases (OpaA and OpaB), transferring the amino group from GABA to pyruvate. While the gabT mutant and the gabT opaA double mutant grew on GABA as a nitrogen source, the final triple mutant did not. The semialdehyde released from GABA by transamination is oxidized by succinate semialdehyde dehydrogenase (GabD). Five of six potential GabD proteins in R. leguminosarum bv. viciae 3841 (GabD1, -D2, -D3, -D4, and -D5) were shown by expression analysis to have this activity. However, only mutations of GabD1, GabD2, and GabD4 were required to prevent utilization of GABA as the sole nitrogen source in culture. The specific enzyme activities of GabT, Opa, and GabD were highly elevated in bacteroids relative to cultured bacteria. This was due to elevated expression of gabT, opaA, gabD1, and gabD2 in nodules. Strains mutated in aminotransferase and succinate semialdehyde dehydrogenases (gabT, opaA, or opaB and gabD1, gabD2, or gabD4, respectively) that cannot use GABA in culture still fixed nitrogen on plants. While GABA catabolism alone is not essential for N2 fixation in bacteroids, it may have a role in energy generation and in bypassing the decarboxylating arm of the tricarboxylic acid cycle.

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Antioxidant defense systems and lipid peroxidation in sideritis taurica's extract-treated diabetic rats

10.1055/s-0037-1603550 ◽

2017 ◽

Author(s):

D Reznikova

Keyword(s):

Lipid Peroxidation ◽

Antioxidant Defense ◽

Diabetic Rats ◽

Defense Systems ◽

Antioxidant Defense Systems

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POSSIBILITIES OF ANTIOXIDANT PROTECTION IN PATIENTS WITH SUPERIOR HYPERPLASIA OF THE PEDESTAL GLAND AND ACCOMPANYING CHRONIC POSTATITIS

СИСТЕМНЫЙ АНАЛИЗ И УПРАВЛЕНИЕ В БИОМЕДИЦИНСКИХ СИСТЕМАХ ◽

10.36622/vstu.2020.19.2.001 ◽

2020 ◽

pp. 8-11

Author(s):

Павел Георгиевич Осипов ◽

Андрей Александрович Береш ◽

Юрий Сергеевич Ханин ◽

Олеся Игоревна Некрылова

Keyword(s):

Lipid Peroxidation ◽

Chronic Prostatitis ◽

Antioxidant Defense ◽

Prostate Gland ◽

Antioxidant Defense System ◽

Complex Treatment ◽

Traditional Treatment ◽

Important Place ◽

Antioxidant Defense Systems ◽

Benign Hyperplasia

Несмотря на достигнутые успехи в диагностике и лечении, на сегодняшний день проблема хронического простатита у пациентов с доброкачественной гиперплазией простаты продолжает оставаться актуальной. Тем временем, выздоровление или же стойкая ремиссия хронического простатита наступает только у 30-35% больных, которые получают традиционное лечение. В патогенезе хронического простатита важное место занимают мембранопатологические процессы, которые обусловлены активацией перекисного окисления липидов и нарушением состояния антиоксидантной системы защиты. У больных с хроническим простатитом и доброкачественной гиперплазией простаты наблюдается существенное усиление процессов липопероксидации на фоне сниженной функциональной способности антиоксидантных систем защиты организма. Поэтому равновесие в оксидантно-антиоксидантной системе является важным звеном в поддержании гомеостаза и, в частности, при патологии предстательной железы, предопределяет включение в комплексное лечение средств антиоксидантного действия. Перспективным можно считать применение препаратов с высоким содержанием биофлавоноидов и антиоксидантных витаминов. Включение в комплексное лечение таких пациентов препарата Кверцетина приводит к сокращению сроков нормализации клинико-лабораторных проявлений заболевания благодаря восстановлению равновесия между перекисным окислением липидов и состоянием антиоксидантной системы защиты Despite the successes achieved in the diagnosis and treatment, to date, the problem of chronic progression in patients with benign hyperplasia of the growth continues to remain relevant. Meanwhile, recovery or persistent remission of chronic prostatitis occurs only in 30-35% of patients who receive traditional treatment. In the pathogenesis of chronic prostatitis, membrane-pathological processes take an important place, which are caused by the activation of lipid peroxidation and impaired state of the antioxidant defense system. In patients with chronic prostate and benign hyperplasia, a significant increase in lipoperoxidation processes is observed against the background of a reduced functional ability of antioxidant defense systems. Therefore, the equilibrium in the oxidant-antioxidant system is an important link in the maintenance of homeostasis and, in particular, with the pathology of the prostate gland, allows the inclusion of antioxidant drugs in the complex treatment. The use of drugs with a high content of bioflavonoids and antioxidant vitamins can be considered promising. The inclusion of such patients in the complex treatment of the drug Quercetin leads to a reduction in the normalization period of the clinical and laboratory manifestations of the disease due to the restoration of the equilibrium between the peroxidation of the lipid peroxidation system and the state of lipid peroxidation

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Pathogenetic Therapy of Patients with Recurrent Genital Herpes

Antibiotics and Chemotherapy ◽

10.37489/0235-2990-2020-65-5-6-30-34 ◽

2020 ◽

Vol 65 (5-6) ◽

pp. 30-34

Author(s):

L. L. Logvina ◽

D. N. Bayram ◽

Z. A. Kambachokova ◽

F. V. Shavaeva ◽

Z. S. Krymshokalova ◽

...

Keyword(s):

Lipid Peroxidation ◽

Blood Plasma ◽

Genital Herpes ◽

Antioxidant Defense ◽

Clinical Manifestations ◽

Complex Treatment ◽

Herpesvirus Infection ◽

Recurrent Genital Herpes ◽

Complex Therapy ◽

Pathogenetic Therapy

Evaluation of the effectiveness of aminophthalhydroside in the complex treatment of patients with recurrent genital herpes was carried out. The efficacy of the drug was assessed by clinical, biochemical, and immunological criteria. The inclusion of aminophthalhydrazide in the complex therapy of patients with recurrent herpesvirus infection led to an earlier relief of clinical manifestations of the disease, an increase in the relapse period, a decrease in lipid peroxidation products in the blood plasma, an increase in the content of antioxidant defense components, and correction of immunological disorders.

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AB0877 Dynamics of Parameters Changes of Antioxidant Defense and Lipid Peroxidation in Patients with Juvenile Idiopathic Arthritis

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2016-eular.4745 ◽

2016 ◽

Vol 75 (Suppl 2) ◽

pp. 1202.1-1202

Author(s):

N. Panko ◽

I. Khadzhynova ◽

S. Shukrey

Keyword(s):

Lipid Peroxidation ◽

Juvenile Idiopathic Arthritis ◽

Antioxidant Defense

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When the Balance Tips: Dysregulation of Mitochondrial Dynamics as a Culprit in Disease

International Journal of Molecular Sciences ◽

10.3390/ijms22094617 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4617

Author(s):

Styliana Kyriakoudi ◽

Anthi Drousiotou ◽

Petros P. Petrou

Keyword(s):

Insulin Resistance ◽

Mitochondrial Function ◽

Human Disease ◽

Molecular Mechanisms ◽

Mitochondrial Dynamics ◽

Mitochondrial Fusion ◽

Mitochondrial Morphology ◽

Disease Development ◽

Pathological Conditions ◽

Wide Range

Mitochondria are dynamic organelles, the morphology of which is tightly linked to their functions. The interplay between the coordinated events of fusion and fission that are collectively described as mitochondrial dynamics regulates mitochondrial morphology and adjusts mitochondrial function. Over the last few years, accruing evidence established a connection between dysregulated mitochondrial dynamics and disease development and progression. Defects in key components of the machinery mediating mitochondrial fusion and fission have been linked to a wide range of pathological conditions, such as insulin resistance and obesity, neurodegenerative diseases and cancer. Here, we provide an update on the molecular mechanisms promoting mitochondrial fusion and fission in mammals and discuss the emerging association of disturbed mitochondrial dynamics with human disease.

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miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00538.2017 ◽

2019 ◽

Vol 316 (3) ◽

pp. H710-H721 ◽

Cited By ~ 6

Author(s):

Victoria L. Nasci ◽

Sandra Chuppa ◽

Lindsey Griswold ◽

Kathryn A. Goodreau ◽

Ranjan K. Dash ◽

...

Keyword(s):

Lipid Peroxidation ◽

Fatty Acid ◽

Palmitic Acid ◽

Fatty Acid Oxidation ◽

Lipid Content ◽

Mitochondrial Respiration ◽

Acid Oxidation ◽

H9c2 Cells ◽

Cellular Lipid ◽

Transfected Cells

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty acid oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty acid oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and palmitic acid. Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and palmitic acid, and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty acid oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

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