Midazolam anesthesia protects neuronal cells from oxidative stress-induced death via activation of the JNK-ERK pathway.
Mol Med Rep
; 15(1): 169-179, 2017 Jan.
Article
en En
| MEDLINE
| ID: mdl-27959401
ABSTRACT
Midazolam is an anesthetic agent commonly used during clinical and surgical procedures, which has been shown to exert ROSsuppressing and apoptosismodulating pharmacological activities in various cellular systems. However, the effects of midazolam on oxidative stress in neuronal cells require elucidation. The present study investigated the effects of midazolam on buthionine sulfoximine (BSO) and hydrogen peroxide (H2O2)induced oxidative stress in primary cortical neuronal cells. In addition, the effects of midazolam on middle cerebral artery occlusion (MCAO) in mice and on ethanolinduced neuroapoptosis in the brains of neonatal mice were determined. Subsequently, cell viability was detected using the MTT assay; intracellular reactive oxygen species (ROS) generation was determined using the 2',7'dichlorodihydrofluorescein diacetate method with confocal microscopy; terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was conducted to detect apoptotic cells; immunohistochemistry was performed to detect activated caspase3; neuronal deficit and infarct volume analyses were conducted; and quantitative polymerase chain reaction and western blotting were performed to detect the expression levels of genes and proteins associated with apoptosis and cell survival pathways. The results demonstrated that BSO (10 mM) and H2O2 (1 mM) suppressed proliferation of cortical neuronal cells by inducing apoptosis. These effects were suppressed following treatment with midazolam in a dosedependent manner. In addition, BSO and H2O2 induced ROS generation in neuronal cells; however, this was effectively suppressed by midazolam (100 µM). Beneficial synergistic effects were detected when midazolam was used in combination with the known antioxidant trolox. BSO and H2O2 also suppressed the protein expression levels of cJun Nterminal kinases (JNK), phosphorylated (p)JNK, extracellular signalregulated kinases (ERK)1/2, pERK1/2, AKT and nuclear factorκB; however, expression was recovered following treatment with midazolam. Midazolam also activated protein kinase Cε, which was suppressed by BSO, in cortical neuronal cells. In MCAO mice, midazolam postconditioning significantly suppressed infarct size and reduced the number of TUNELpositive cells. In addition, the expression levels of caspase3 and poly (ADPribose) polymerase were suppressed in a dosedependent manner. In neonatal mice, midazolam reduced ethanolinduced activated caspase3 staining and apoptotic TUNEL staining. The results of the present study demonstrated that midazolam may protect against neuronal degeneration and neuroapoptosis induced by physiological and oxidative stress.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Midazolam
/
Muerte Celular
/
Fármacos Neuroprotectores
/
Sistema de Señalización de MAP Quinasas
/
Adyuvantes Anestésicos
/
Neuronas
Límite:
Animals
Idioma:
En
Revista:
Mol Med Rep
Año:
2017
Tipo del documento:
Article