Critical Care Medicine
Relationship between M2-type microglia-derived exosomes and neuronal oxygen-glucose deprivation and restoration injury in mice
Shan Kaiyue, Li Huimin, Wang Hong, Chen Jingyan, Liu Wenjie, Chen Huailong, Zhang Gaofeng, Wang Mingshan, Dong Rui
Published 2022-10-20
Cite as Chin J Anesthesiol, 2022, 42(10): 1233-1237. DOI: 10.3760/cma.j.cn131073.20220413.01016
Abstract
ObjectiveTo evaluate the relationship between M2-type microglia-derived exosomes (M2-exo) and neuronal oxygen-glucose deprivation and restoration (OGD/R) injury in mice.
MethodsMouse neuroblastoma cells (N2a cells) and BV2 microglia were cultured in vitro, and BV2 microglia were activated to M2 type using 20 ng/ml IL-4, and M0-type microglia-derived exosomes (M0-exo) and M2-exo were extracted.N2a cells were divided into 4 groups (n=23 each) using the random number table method: control+ M0-exo group (C+ M0 group), control+ M2-exo group (C+ M2 group), OGD/R+ M0-exo group (O+ M0 group) and OGD/R+ M2-exo group (O+ M2 group).M0-exo and M2-exo (final concentration 100 μg/ml) were added in C+ M0 and C+ M2 groups, respectively, and the cells were incubated for 24 h. M0-exo and M2-exo (final concentration 100 μg/ml) were added at 3 h after oxygen and glucose deprivation, and then the cells were incubated for 24 h in O+ M0 and O+ M2 groups, respectively.N2a cell viability was measured by the CCK-8 method, and the severity of cell damage was assessed using the lactic dehydrogenase (LDH) release rate.The expression of Bax and Bcl-2 protein and mRNA was detected by quantitative real-time polymerase chain reaction and Western blot.
ResultsCompared with C+ M0 group, no significant changes were found in N2a cell viability, LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio in C+ M2 group (P>0.05), and N2a cell viability was significantly decreased, and the LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio were increased in O+ M0 group (P<0.05).Compared with C+ M2 group, the N2a cell viability was significantly decreased, and the LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio were increased in O+ M2 group (P<0.05).Compared with O+ M0 group, N2a cell viability was significantly increased, and LDH release rate, Bax/Bcl-2 ratio, and Bax mRNA/Bcl-2 mRNA ratio were decreased in O+ M2 group (P<0.05).
ConclusionsM2-exo exerts an endogenous protective effect during OGD/R in mouse neurons, which may be related to the inhibition of cell apoptosis.
Key words:
Microglia; Exosomes; Neurons; Reperfusion injury
Contributor Information
Shan Kaiyue
Graduate School of Dalian Medical University, Dalian 116044, China
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Li Huimin
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Wang Hong
Department of Education and Training, Qingdao Women and Children′s Hospital, Qingdao University, Qingdao 266071, China
Chen Jingyan
Department of Anesthesiology, Qingdao Medical College of Nanjing Medical University, Qingdao 266071, China
Liu Wenjie
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Chen Huailong
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Zhang Gaofeng
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Wang Mingshan
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Dong Rui
Department of Anesthesiology, Qingdao Municipal Hospital Affiliated to Qingdao University, Qingdao 266071, China
Medical School, Nanjing University, Nanjing 210093, China
