Erythrocyte deformability in an in vitro model of hypoxia/reoxygenation – protective effects of superoxide dismutase and catalase

Ischemic heart disease (IHD) is a cardiovascular disease with high fatality rate, and its pathogenesis is closely related to oxidative stress. Reactive oxygen species (ROS) in oxidative stress can lead to myocardial ischemia (MI) injury in many ways. Therefore, the application of antioxidants may be an effective way to prevent IHD. In recent years, glutathione peroxidase 4 (GPx4) has received increasing attention due to its antioxidant effect. In a previous study, we used the new chimeric tRNAUTuT6 to express highly active recombinant human GPx4 (rhGPx4) in amber-less Escherichia coli. In this study, we established an isoproterenol- (ISO-) induced MI injury model in rats and an in vitro model to research the protective effect and mechanism of rhGPx4 on MI injury. The results showed that rhGPx4 could reduce the area of myocardial infarction and ameliorate the pathological injury of heart tissue, significantly reduce ISO-induced abnormalities on electrocardiogram (ECG) and cardiac serum biomarkers, protect mitochondrial function, and attenuate cardiac oxidative stress injury. In an in vitro model, the results also confirmed that rhGPx4 could inhibit ISO-induced oxidative stress injury and cardiomyocyte apoptosis. The mechanism of action of rhGPx4 involves not only the inhibition of lipid peroxidation by eliminating ROS but also keeping a normal level of endogenous antioxidant enzymes by eliminating ROS, thereby preventing oxidative stress injury in cardiomyocytes. Additionally, rhGPx4 could inhibit cardiomyocyte apoptosis through a mitochondria-dependent pathway. In short, rhGPx4, a recombinant antioxidant enzyme, can play an important role in the prevention of IHD and may have great potential for application.

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Cannabidiol exerts protective effects in an in vitro model of Parkinson's disease activating AKT/mTOR pathway

Fitoterapia ◽

10.1016/j.fitote.2020.104553 ◽

2020 ◽

Vol 143 ◽

pp. 104553 ◽

Cited By ~ 3

Author(s):

Agnese Gugliandolo ◽

Federica Pollastro ◽

Placido Bramanti ◽

Emanuela Mazzon

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

In Vitro Model ◽

Mtor Pathway ◽

Protective Effects ◽

Vitro Model

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Low-dose lipopolysaccharide protects nerve cells against spinal cord injury via regulating the PI3K–AKT–Nrf2 signaling pathway

Biochemistry and Cell Biology ◽

10.1139/bcb-2020-0641 ◽

2021 ◽

pp. 1-9

Author(s):

Wei-Chao Li ◽

Shao-Ping Yao ◽

Jun Zhang ◽

Wei-Bing Liu ◽

Jie Liu ◽

...

Keyword(s):

Pc12 Cells ◽

In Vitro Model ◽

Low Dose ◽

Nuclear Translocation ◽

Protective Effects ◽

Expression Levels ◽

Nrf2 Signaling Pathway ◽

Cord Injury ◽

Vitro Model

This study explored the molecular mechanism behind the protective effects from low-dose lipopolysaccharide (LPS) on an in-vitro model of spinal cord injury (SCI). For this, PC12 cells were treated with different concentrations of LPS and the cell counting kit-8 assay was used to measure the toxicity of LPS to the cells. Next, we used immunofluorescence to measure nuclear translocation of Nrf2 in PC12 cells. PC12 cells were then treated with IGF-1 (PI3K agonist) and LY294002 (PI3K inhibitor). An in-vitro model of SCI was then established via oxygen–glucose deprivation/reoxygenation. Rates of apoptosis were measured using flow cytometry and the TUNEL assay. Low-dose LPS increased the expression levels of Nrf2, p-PI3K/PI3K, and p-AKT/AKT, and facilitated nuclear translocation of Nrf2. The activation of PI3K–AKT signaling by IGF-1 significantly increased the expression of Nrf2, whereas inhibition of PI3K–AKT signaling significantly decreased the expression of Nrf2. Low-dose LPS reduced the apoptotic ratio of PC12 cells, decreased the expression levels of caspase 3 and caspase 9, and increased the expression levels of HO-1, NQO1, and γ-GCS. Low-dose LPS also reduced the rate of apoptosis and oxidative stress by activating the PI3K–AKT–Nrf2 signaling pathway. Collectively, the results indicate that PI3K–AKT–Nrf2 signaling participates in the protective effects from low-dose LPS in an in-vitro PC12 cell model of SCI.

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Glutamate as a potential “survival factor” in an in vitro model of neuronal hypoxia/reoxygenation injury: leading role of the Na+/Ca2+ exchanger

Cell Death and Disease ◽

10.1038/s41419-018-0784-6 ◽

2018 ◽

Vol 9 (7) ◽

Cited By ~ 18

Author(s):

Silvia Piccirillo ◽

Pasqualina Castaldo ◽

Maria Loredana Macrì ◽

Salvatore Amoroso ◽

Simona Magi

Keyword(s):

In Vitro Model ◽

Survival Factor ◽

Leading Role ◽

Reoxygenation Injury ◽

Vitro Model ◽

Hypoxia Reoxygenation

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Sangiovese cv Pomace Seeds Extract-Fortified Kefir Exerts Anti-Inflammatory Activity in an In Vitro Model of Intestinal Epithelium Using Caco-2 Cells

Antioxidants ◽

10.3390/antiox9010054 ◽

2020 ◽

Vol 9 (1) ◽

pp. 54 ◽

Cited By ~ 1

Author(s):

Gabriele Carullo ◽

Paolo Governa ◽

Umile Gianfranco Spizzirri ◽

Marco Biagi ◽

Fabio Sciubba ◽

...

Keyword(s):

In Vitro Model ◽

Fermented Milk ◽

Food Allergies ◽

Protective Effects ◽

Electric Resistance ◽

Nutrition Science ◽

Fortified Milk ◽

Inflammatory Bowel ◽

Vitro Model

Inflammatory bowel disease and food allergies are a growing topic in the field of nutrition science. Polyphenols, which are the most important secondary metabolites of plants, demonstrated to modulate the expression and/or production of numerous proteins, but also to regulate the intestinal ecosystem. In this context, our aim was the investigation of protective effects against the gastrointestinal mucosa of fortified milk kefir obtained by adding seeds extract from Sangiovese cv. Pomace. Methods: An ultrasound-assisted method was used to obtain the extracts. All the extracts were assayed for the antioxidant activity. The best extract was used as an additive of fermented milk kefir to obtain a fortified final product. Kefir samples were analyzed by NMR spectroscopy. The efficiency of the barrier functions was evaluated by measuring trans-epithelial electric resistance (TEER) using a voltmeter. Results: the enriched kefir (Ksgn) possesses higher antioxidant performances compared to the unfortified sample (Kwht). Kwht and Ksgn did not alter Caco-2 TEER in basal condition.

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Development of an in vitro model for study of the efficacy of ischemic preconditioning in human skeletal muscle against ischemia-reperfusion injury

Journal of Applied Physiology ◽

10.1152/japplphysiol.00278.2006 ◽

2006 ◽

Vol 101 (5) ◽

pp. 1335-1342 ◽

Cited By ~ 14

Author(s):

Glyka Martou ◽

Catherine A. O’Blenes ◽

Ning Huang ◽

Sandra E. McAllister ◽

Peter C. Neligan ◽

...

Keyword(s):

Skeletal Muscle ◽

Protective Effect ◽

Ischemic Preconditioning ◽

Human Skeletal Muscle ◽

In Vitro Model ◽

Ischemia Reperfusion ◽

Mtt Reduction ◽

Vitro Model ◽

Hypoxia Reoxygenation

Ischemia-reperfusion (I/R) injury causes skeletal muscle infarction and ischemic preconditioning (IPC) augments ischemic tolerance in animal models. To date, this has not been demonstrated in human skeletal muscle. This study aimed to develop an in vitro model to investigate the efficacy of simulated IPC in human skeletal muscle. Human skeletal muscle strips were equilibrated in oxygenated Krebs-Henseleit-HEPES buffer (37°C). Aerobic and reperfusion phases were simulated by normoxic incubation and reoxygenation, respectively. Ischemia was simulated by hypoxic incubation. Energy store, cell viability, and cellular injury were assessed using ATP, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2 H-tetrazolium bromide (MTT), and lactate dehydrogenase (LDH) assays, respectively. Morphological integrity was assessed using electron microscopy. Studies were designed to test stability of the preparation ( n = 5–11) under normoxic incubation over 24 h; the effect of 1, 2, 3, 4, or 6 h hypoxia followed by 2 h of reoxygenation; and the protective effect of hypoxic preconditioning (HPC; 5 min of hypoxia/5 min of reoxygenation) before 3 h of hypoxia/2 h of reoxygenation. Over 24 h of normoxic incubation, muscle strips remained physiologically intact as assessed by MTT, ATP, and LDH assays. After 3 h of hypoxia/2 h of reoxygenation, MTT reduction levels declined to 50.1 ± 5.5% ( P < 0.05). MTT reduction levels in HPC (82.3 ± 10.8%) and normoxic control (81.3 ± 10.2%) groups were similar and higher ( P < 0.05) than the 3 h of hypoxia/2 h of reoxygenation group (45.2 ± 5.8%). Ultrastructural morphology was preserved in normoxic and HPC groups but not in the hypoxia/reoxygenation group. This is the first study to characterize a stable in vitro model of human skeletal muscle and to demonstrate a protective effect of HPC in human skeletal muscle against hypoxia/reoxygenation-induced injury.

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