Critical Care Medicine
Role of EAAT2 in cannabinoid receptor 2 activation-induced attenuation of microglial injury caused by glutamate
Xiajing Zhang, Yan Feng, Liying Tian, Jing Wang, Yongping Shao, Xude Sun
Published 2018-08-20
Cite as Chin J Anesthesiol, 2018, 38(8): 1009-1011. DOI: 10.3760/cma.j.issn.0254-1416.2018.08.029
Abstract
ObjectiveTo evaluate the role of excitatory amino acid transporter 2 (EAAT2) in cannabinoid receptor 2 (CB2 receptor) activation-induced attenuation of microglial injury caused by glutamate.
MethodsN9 microglial cells were divided into 4 groups (n=26 each) using a random number table method: control group (group Con), glutamate group (group Glu), CB2 receptor agonist AM1241 plus glutamate group (group AM1241+ Glu) and AM1241 plus EAAT inhibitor TBOA plus glutamate group (group AM1241+ TBOA+ Glu). The cells were routinely cultured for 30 h in group Con.In group Glu, the cells were routinely cultured for 6 h, and then were incubated for 24 h in the culture medium containing glutamate 10 mmol/L.In group AM1241+ Glu, the cells were incubated for 4 h in the culture medium containing AM1241 2 μmol/L, and then were routinely cultured for 2 h, and then were incubated for 24 h in the culture medium containing glutamate 10 mmol/L.In group AM1241+ TBOA+ Glu, the cells were incubated for 4 h in the culture medium containing AM1241 2 μmol/L and TBOA 100 μmol/L, and then were routinely cultured for 2 h, and then were incubated for 24 h in the culture medium containing glutamate 10 mmol/L.The cell viability was measured by MTT assay, the activity of lactic dehydrogenase (LDH) in supernatant was determined using colorimetric method, and the expression of EAAT2 was determined by Western blot.
ResultsCompared with group Con, the cell viability was significantly decreased and LDH activity was increased in Glu, AM1241+ Glu and AM1241+ TBOA+ Glu groups, and the expression of EAAT2 was significantly up-regulated in Glu and AM1241+ Glu groups (P<0.05). Compared with group Glu, the cell viability was significantly increased, LDH activity was decreased, and the expression of EAAT2 was up-regulated in group AM1241+ Glu (P<0.05), and no significant change was found in the parameters mentioned above in group AM1241+ TBOA+ Glu (P>0.05). Compared with group AM1241+ Glu, the cell viability was significantly decreased, LDH activity was increased, and the expression of EAAT2 was down-regulated in group AM1241+ TBOA+ Glu (P<0.05).
ConclusionThe mechanism by which the activation of CB2 receptor attenuates microglial injury caused by glutamate is related to up-regulating the expression of EAAT2.
Key words:
Excitatory amino acid transporter 2; Receptor, cannabinoid, CB2; Glutamic acid; Microglia
Contributor Information
Xiajing Zhang
Department of Anesthesiology, Xi′an No.4 Hospital, Xi′an 710001, China
Yan Feng
Department of Anesthesiology, Xi′an No.4 Hospital, Xi′an 710001, China
Liying Tian
Department of Anesthesiology, Tangdu Hospital, Air Force Military Medical University Xi′an, 710038, China
Jing Wang
Department of Anesthesiology, Tangdu Hospital, Air Force Military Medical University Xi′an, 710038, China
Yongping Shao
Department of Anesthesiology, Xi′an No.4 Hospital, Xi′an 710001, China
Xude Sun
Department of Anesthesiology, Tangdu Hospital, Air Force Military Medical University Xi′an, 710038, China
