M2型小胶质细胞源性外泌体与小鼠神经元氧糖剥夺/复糖复氧损伤的关系  

Relationship between M2-type microglia-derived exosomes and neuronal oxygen-glucose deprivation and restoration injury in mice

作　　者：单凯悦 李慧敏 王红[3] 陈静燕 刘文洁 陈怀龙[2] 张高峰 王明山[2] 董瑞 Shan Kaiyue;Li Huimin;Wang Hong;Chen Jingyan;Liu Wenjie;Chen Huailong;Zhang Gaofeng;Wang Mingshan;Dong Rui(Graduate School of Dalian Medical University,Dalian 116044,China;Department of Anesthesiology,Qingdao Municipal Hospital Affiliated to Qingdao University,Qingdao 266071,China;Department of Education and Training,Qingdao Women and Children′s Hospital,Qingdao University,Qingdao 266071,China;Department of Anesthesiology,Qingdao Medical College of Nanjing Medical University,Qingdao 266071,China;Medical School,Nanjing University,Nanjing 210093,China)

机构地区：[1]大连医科大学研究生院,大连116044 [2]青岛大学附属青岛市市立医院麻醉科,青岛266071 [3]青岛大学附属青岛市妇女儿童医院教育培训科,青岛266034 [4]南京医科大学青岛临床医学院青岛市市立医院麻醉科,青岛266071 [5]南京大学医学院,南京210093

出　　处：《中华麻醉学杂志》2022年第10期1233-1237,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82001132);贝朗麻醉科研基金(BBDF-2019-010)。

摘　　要：目的评价M2型小胶质细胞源性外泌体(M2-exo)与小鼠神经元氧糖剥夺/复糖复氧(OGD/R)损伤的关系。方法体外培养小鼠神经母细胞(N2a细胞)及BV2小胶质细胞,采用20 ng/ml IL-4将BV2小胶质细胞激活为M2型,提取M0型小胶质细胞源性外泌体(M0-exo)和M2-exo。采用随机数字表法将N2a细胞分为4组(n=23):对照+M0-exo组(C+M0组)、对照+M2-exo组(C+M2组)、OGD/R+M0-exo组(O+M0组)和OGD/R+M2-exo组(O+M2组)。C+M0组和C+M2组分别加入M0-exo和M2-exo(终浓度100μg/ml)孵育24 h,O+M0组和O+M2组于氧糖剥夺3 h时分别加入M0-exo和M2-exo(终浓度100μg/ml),再复糖复氧24 h。采用CCK-8法检测N2a细胞活力,采用LDH释放率评估细胞损伤程度,采用Western blot法及实时q-PCR检测Bax和Bcl-2及其mRNA表达。结果与C+M0组比较,C+M2组N2a细胞活力、LDH释放率、Bax/Bcl-2比值和Bax mRNA/Bcl-2 mRNA比值差异无统计学意义(P>0.05),O+M0组N2a细胞活力下降,LDH释放率、Bax/Bcl-2比值和Bax mRNA/Bcl-2 mRNA比值升高(P<0.05)。与C+M2组比较,O+M2组N2a细胞活力下降,LDH释放率、Bax/Bcl-2比值和Bax mRNA/Bcl-2 mRNA比值升高(P<0.05)。与O+M0组比较,O+M2组N2a细胞活力升高,LDH释放率、Bax/Bcl-2比值和Bax mRNA/Bcl-2 mRNA比值降低(P<0.05)。结论M2-exo在小鼠神经元OGD/R过程中发挥内源性保护作用,可能与抑制细胞凋亡有关。Objective To evaluate the relationship between M2-type microglia-derived exosomes(M2-exo)and neuronal oxygen-glucose deprivation and restoration(OGD/R)injury in mice.Methods Mouse 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 100μg/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 100μg/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.Results Compared 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).Conclusions M2-exo exerts an endogenous protective effect during OGD/R in mouse neurons,which may be related to the inhibition of cell apoptosis.

关 键 词：小神经胶质细胞 外泌体 神经元 再灌注损伤 

分 类 号：R614[医药卫生—麻醉学]