TY - JOUR
T1 - Astrocyte-conditioned medium attenuates glutamate-induced apoptotic cell death in primary cultured spinal cord neurons of rats
AU - Lu, Xiaojian
AU - Al-Aref, Rami
AU - Zhao, Dong
AU - Shen, Jianhong
AU - Yan, Yaohua
AU - Gao, Yilu
N1 - Publisher Copyright:
© W. S. Maney & Son Ltd 2015.
PY - 2015/7/20
Y1 - 2015/7/20
N2 - Objectives: To better understand the neuroprotective role of astrocytes in spinal cord injury (SCI), we investigated whether astrocyte-conditioned medium (ACM) can attenuate glutamate-induced apoptotic cell death in primary cultured spinal cord neurons. Methods: Spinal cord neurons were pretreated with ACM for 24 hours. Subsequently, they were exposed to glutamate (125 mM) for 1 hour. The neurons were then incubated for 24 hours. Following that, measurements assessing cell viability and lactate dehydrogenase (LDH) release were performed. Apoptosis was confirmed through cell morphology using Hoechst 33342 staining and terminal deoxynucleotidyl transferase dUTP-mediated nicked end labeling (TUNEL) assay. Assessment for expression of apoptotic enzymes, including Caspase-3, Bcl-2 and Bax, was performed using Western Blot Analysis. Results: Astrocyte-conditioned medium pretreatment of neurons showed both an increase in spinal cord neuron viability and a decrease in LDH release in a dose-dependent pattern. Moreover, pretreatment seems to attenuate glutamate-induced apoptotic cell death, antagonise glutamate-induced up-regulation of Caspase-3 expression and downregulate Bcl-2/Bax protein expression ratio. Conclusions: By attenuating glutamate-induced apoptotic cell death in primary cultured spinal cord neurons of rats, ACM seems to provide a neuroprotective effect by regulating apoptosis-related protein expression. Our results provide an experimental basis for clinical applications and potential therapeutic use of ACM in SCI.
AB - Objectives: To better understand the neuroprotective role of astrocytes in spinal cord injury (SCI), we investigated whether astrocyte-conditioned medium (ACM) can attenuate glutamate-induced apoptotic cell death in primary cultured spinal cord neurons. Methods: Spinal cord neurons were pretreated with ACM for 24 hours. Subsequently, they were exposed to glutamate (125 mM) for 1 hour. The neurons were then incubated for 24 hours. Following that, measurements assessing cell viability and lactate dehydrogenase (LDH) release were performed. Apoptosis was confirmed through cell morphology using Hoechst 33342 staining and terminal deoxynucleotidyl transferase dUTP-mediated nicked end labeling (TUNEL) assay. Assessment for expression of apoptotic enzymes, including Caspase-3, Bcl-2 and Bax, was performed using Western Blot Analysis. Results: Astrocyte-conditioned medium pretreatment of neurons showed both an increase in spinal cord neuron viability and a decrease in LDH release in a dose-dependent pattern. Moreover, pretreatment seems to attenuate glutamate-induced apoptotic cell death, antagonise glutamate-induced up-regulation of Caspase-3 expression and downregulate Bcl-2/Bax protein expression ratio. Conclusions: By attenuating glutamate-induced apoptotic cell death in primary cultured spinal cord neurons of rats, ACM seems to provide a neuroprotective effect by regulating apoptosis-related protein expression. Our results provide an experimental basis for clinical applications and potential therapeutic use of ACM in SCI.
KW - Apoptosis
KW - Astrocyte-conditioned medium
KW - Glutamate
KW - Spinal cord neurons
UR - http://www.scopus.com/inward/record.url?scp=84937542068&partnerID=8YFLogxK
U2 - 10.1179/1743132815Y.0000000059
DO - 10.1179/1743132815Y.0000000059
M3 - Article
C2 - 26038835
AN - SCOPUS:84937542068
SN - 0161-6412
VL - 37
SP - 803
EP - 808
JO - Neurological Research
JF - Neurological Research
IS - 9
ER -