scholarly journals Mechanistic Distinctions between Excitotoxic and Acidotic Hippocampal Damage in an in vitro Model of Ischemia

1990 ◽  
Vol 10 (4) ◽  
pp. 527-535 ◽  
Author(s):  
Geoffrey C. Tombaugh ◽  
Robert M. Sapolsky
Keyword(s):  
Nmda Receptor ◽  
Cell Injury ◽  
Neuronal Damage ◽  
Nmda Receptor Antagonist ◽  
Hippocampal Damage ◽  
Neuron Death ◽  
Vitro Model ◽  
Posthypoxic Period ◽  
Ph Range

Excitotoxicity is believed to underlie the selective loss of vulnerable neurons after transient ischemia, while lactic acidosis seems to be the principal feature and probable cause of tissue infarcts. Primary hippocampal cultures containing both neurons and astrocytes derived from fetal rats were used to examine the relative contributions of and interactions between excitotoxic and acidotic cell injury. Hypoxia-induced damage was energy dependent and involved the N-methyl-D-aspartate (NMDA) receptor. Glucose above 1 m M could completely protect against hypoxia-induced injury in a pH range of 7.4–6.5, while the NMDA receptor antagonist D,L-2-amino-5-phosphonovaleric acid (500 μ M) during the posthypoxic period provided only partial protection in the absence of glucose. Astrocyte cultures were undamaged by ischemic-like treatment in this pH range, suggesting that hypoxia-induced cell death in mixed cultures was restricted to neurons. Lowering the extracellular pH to 7.0 and 6.5 caused no neuronal damage in normoxic controls, but in each case provided significant protection against hypoxic neuronal injury. In contrast, a second type of neurotoxicity was observed after a 6-h exposure to pH 6.0, while exposure to pH 5.5 was required to kill astrocytes. This acidotic damage appeared to be energy independent and did not involve the NMDA receptor. These results suggest that excitotoxic neuron death has an energetic component and that acidosis may produce both protective and damaging effects in the hippocampus during ischemic insults.

Chronic neocortical epileptogenesis in vitro

1994 ◽  
Vol 71 (5) ◽  
pp. 1762-1773 ◽  
Author(s):  
S. N. Hoffman ◽  
P. A. Salin ◽  
D. A. Prince
Keyword(s):  
White Matter ◽  
Current Source ◽  
Nmda Receptor Antagonist ◽  
White Matter Injury ◽  
Posttraumatic Epilepsy ◽  
Layer V ◽  
Vitro Model ◽  
Interictal Discharges ◽  
Interictal Discharge

1. We used an in vitro model to explore critical aspects of chronic epileptogenesis. Partial neocortical isolations having intact blood supply were made in rat and guinea pig from postnatal day 7 to 34 and then examined 1 to 150 days later in standard brain slice preparations. 2. The epileptogenic potential of several different types of lesions was assessed. Slices containing transcortical (i.e., gray matter) lesions, with or without a contiguous white matter injury (i.e., “undercut”), developed chronic epileptogenesis after a latency of approximately 1–2 wk, manifested by evoked and spontaneous “interictal” discharges and evoked “ictal” events. The region of hyperexcitability did not extend beyond approximately 2 mm from the chronic transcortical lesion and was rarely observed in slices having only an apparent white matter injury. 3. Multiple recordings and current source density (CSD) analysis identified layer V as the source of the interictal discharge. 4. Significant differences in CSD profiles of the evoked interictal discharge occurred between chronically epileptogenic slices and control (noninjured) slices bathed in the convulsant, bicuculline methiodide, suggesting that mechanisms other than disinhibition must be involved in posttraumatic epileptogenesis. 5. Interictal events were blocked in most but not all chronically injured slices by application of the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonovalerate (D-AP5), suggesting that non-NMDA receptors were predominantly involved in some preparations. 6. This model of chronic epileptogenesis in vitro will be useful in studies relevant to mechanisms of posttraumatic epilepsy in man.

scholarly journals T-2 Toxin Exposure Induces Apoptosis in TM3 Cells by Inhibiting Mammalian Target of Rapamycin/Serine/Threonine Protein Kinase(mTORC2/AKT) to Promote Ca2+Production

2018 ◽  
Vol 19 (11) ◽  
pp. 3360 ◽  
Author(s):  
Ji Wang ◽  
Chenglin Yang ◽  
Zhihang Yuan ◽  
Jine Yi ◽  
Jing Wu
Keyword(s):  
Cell Injury ◽  
Mammalian Target Of Rapamycin ◽  
Annexin V ◽  
Target Of Rapamycin ◽  
Dependent Manner ◽  
Toxin Exposure ◽  
Concentration Changes ◽  
Vitro Model ◽  
Induced Apoptosis

Although mTOR (the mammalian target of rapamycin) can regulate intracellular free Ca2+concentration in normal cultured podocytes, it remains elusive as to how mTORC2/AKT-mediated Ca2+participates in the process of T-2 toxin-induced apoptosis. The potential signaling responsible for intracellular Ca2+ concentration changes was investigated using immunoblot assays in an in vitro model of TM3 cell injury induced by T-2 toxin. Changes in Ca2+ were assessed using the Ca2+-sensitive fluorescent indictor dye Fura 2-AM. The cytotoxicity of TM3 cells was assessed with an MTT bioassay, and apoptosis was measured using Annexin V-FITC staining. Following T-2 toxin treatment, the growth of cells, phospho-mTORSer2481, phospho-mTORSer2448, and phospho-AktSer473 were significantly decreased in a time-dependent manner, whereas Ca2+ and apoptosis were increased. T-2 toxin-induced apoptosis was prevented by BAPTA-AM (a Ca2+chelator) and MHY1485 (an mTOR activator), and the application of mTOR activator MHY1485 also prevented the increase of intracellular free Ca2+concentration in TM3 cells. Our results strongly suggest that T-2 toxin exposure induces apoptosis in TM3 cells by inhibiting mTORC2/AKT to promote Ca2+ production.

Kidney tubular epithelial cell injury in shock wave lithoripsy: The search for an appropriate in vitro model

1995 ◽  
Vol 98 (5) ◽  
pp. 2942-2943
Author(s):  
James A. McAteer ◽  
Andrew P. Evan ◽  
James E. Lingeman ◽  
Sharon P. Andreoli
Keyword(s):  
Shock Wave ◽  
Epithelial Cell ◽  
In Vitro Model ◽  
Cell Injury ◽  
Tubular Epithelial Cell ◽  
Vitro Model ◽  
Epithelial Cell Injury

scholarly journals No Cytotoxic and Inflammatory Effects of Empagliflozin and Dapagliflozin on Primary Renal Proximal Tubular Epithelial Cells under Diabetic Conditions In Vitro

2020 ◽  
Vol 21 (2) ◽  
pp. 391 ◽  
Author(s):  
Patrick C. Baer ◽  
Benjamin Koch ◽  
Janina Freitag ◽  
Ralf Schubert ◽  
Helmut Geiger
Keyword(s):  
Epithelial Cells ◽  
Cell Injury ◽  
Cellular Damage ◽  
Tubular Epithelial Cells ◽  
Urinary Glucose ◽  
Proximal Tubular Epithelial Cells ◽  
Proximal Tubular ◽  
Vitro Model ◽  
Renal Proximal Tubular

Gliflozins are inhibitors of the renal proximal tubular sodium-glucose co-transporter-2 (SGLT-2), that inhibit reabsorption of urinary glucose and they are able to reduce hyperglycemia in patients with type 2 diabetes. A renoprotective function of gliflozins has been proven in diabetic nephropathy, but harmful side effects on the kidney have also been described. In the current project, primary highly purified human renal proximal tubular epithelial cells (PTCs) have been shown to express functional SGLT-2, and were used as an in vitro model to study possible cellular damage induced by two therapeutically used gliflozins: empagliflozin and dapagliflozin. Cell viability, proliferation, and cytotoxicity assays revealed that neither empagliflozin nor dapagliflozin induce effects in PTCs cultured in a hyperglycemic environment, or in co-medication with ramipril or hydro-chloro-thiazide. Oxidative stress was significantly lowered by dapagliflozin but not by empagliflozin. No effect of either inhibitor could be detected on mRNA and protein expression of the pro-inflammatory cytokine interleukin-6 and the renal injury markers KIM-1 and NGAL. In conclusion, empa- and dapagliflozin in therapeutic concentrations were shown to induce no direct cell injury in cultured primary renal PTCs in hyperglycemic conditions.

Characterization of 3-carboxy-5-phosphono-1,2,3,4-tetrahydroisoquinoline (SC-48981), a potent competitive (NMDA) receptor antagonist, in vitro and in vivo

1992 ◽  
Vol 135 (2) ◽  
pp. 149-152 ◽  
Author(s):  
Michael L. Vazquez ◽  
Danny J. Garland ◽  
Eric T. Sun ◽  
Julie A. Cler ◽  
Steve J. Mick ◽  
...  
Keyword(s):  
Nmda Receptor ◽  
Receptor Antagonist ◽  
Nmda Receptor Antagonist

scholarly journals Progression of Neuronal Damage in an In Vitro Model of the Ischemic Penumbra

PLoS ONE ◽  
2016 ◽  
Vol 11 (2) ◽  
pp. e0147231 ◽  
Author(s):  
Joost le Feber ◽  
Stelina Tzafi Pavlidou ◽  
Niels Erkamp ◽  
Michel J. A. M. van Putten ◽  
Jeannette Hofmeijer
Keyword(s):  
In Vitro Model ◽  
Neuronal Damage ◽  
Ischemic Penumbra ◽  
Vitro Model

scholarly journals Salvianolate lyophilized injection induced autophagy against neuronal apoptosis through the Akt/mTOR pathway in Neuro-2a cells

2020 ◽  
Author(s):  
Dongna Li ◽  
Mengmeng Ma ◽  
Wenqi Zhang ◽  
Yangyang Xu ◽  
Shaoxia Wang ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Cell Injury ◽  
Biological Effects ◽  
Glucose Deprivation ◽  
Novel Therapy ◽  
Substrate Protein ◽  
N2a Cells ◽  
Autophagy Inhibitor ◽  
Vitro Model

Abstract Background: Stroke remains a worldwide health problem. Salvianolate lyophilized injection (SLI) is one of the most widely-used in clinics in China for treating stroke. Our previous research found that SLI protects against stroke by inhibiting oxidative stress and inflammation. Whether SLI has other biological effects on stroke remains unknown. Methods: Neuro-2a (N2a) cells were treated with SLI (10, 25, 50μg/ml) and exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) as an in vitro model of ischemic stroke. Then, cell apoptosis was assessed using flow cytometry, apoptosis and autophagy related proteins were investigated by western blotting. Autophagy inhibitor 3-methyladenine (3-MA) pretreatment was used to detect the apoptosis effect of SLI on N2a cells. Results: In this work, we verified that SLI promoted cell proliferation, inhibited cell injury and apoptosis in OGD/R-induced N2a cells. The increase the expression of autophagy markers LC3 and Beclin-1, and decrease autophagy substrate protein p62 demonstrated the induction of autophagy by SLI. Additionally, we found that SLI activation of autophagy significantly preceded inhibition of apoptosis in N2a. Pretreatment with autophagy inhibitor 3-MA could antagonize the protective effect of SLI on inhibiting apoptosis, which may be related to the Akt/mTOR signalling pathway. Conclusions: The present study indicated that SLI has potential as a novel therapy for ischemic stroke, and its possible mechanism is to induced autophagy and inhibit apoptosis through blocking Akt/mTOR signaling pathway in OGD/R-induced N2a cells, providing new insights into the mechanism of SLI in the treatment of stroke.

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