Involvement of MAPK pathways in NMDA-induced apoptosis of rat cortical neurons.
- Author:
Xiao-Rong YANG
1
,
2
;
Ping SUN
;
Hua-Ping QIN
;
Pei-Pei SI
;
Xue-Fei SUN
;
Ce ZHANG
Author Information
1. Department of Physiology
2. Department of Nutrition of the First Hospital, Shanxi Medical University, Taiyuan 030001, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Anthracenes;
pharmacology;
Apoptosis;
Caspase 3;
metabolism;
Cells, Cultured;
Extracellular Signal-Regulated MAP Kinases;
antagonists & inhibitors;
Imidazoles;
pharmacology;
JNK Mitogen-Activated Protein Kinases;
antagonists & inhibitors;
MAP Kinase Signaling System;
N-Methylaspartate;
pharmacology;
Neurons;
cytology;
Primary Cell Culture;
Pyridines;
pharmacology;
Rats;
p38 Mitogen-Activated Protein Kinases;
antagonists & inhibitors
- From:
Acta Physiologica Sinica
2012;64(6):609-616
- CountryChina
- Language:English
-
Abstract:
NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.
