氧糖剥夺再灌注对星形胶质细胞缝隙连接与糖摄取功能的影响  

Effects of oxygen glucose deprivation and reperfusion on gap junction and glucose uptake of astrocytes

作　　者：张向阳[1] 王翠芳[2] 郭卫春[1] 陈燕[2] Zhang Xiangyang;Wang Cuifang;Guo Weichun;Chen Yan(Department of orthopedics,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Neurology,Renmin,Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院骨科,430060 [2]武汉大学人民医院神经内科,430060

出　　处：《中华行为医学与脑科学杂志》2019年第11期967-970,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金项目(81501053)。

摘　　要：目的探讨氧糖剥夺(oxygen-glucose deprivation,OGD)条件下脑星形胶质细胞缝隙连接与糖摄取功能之间关系。方法取出生1 d的SD大鼠大脑皮层进行星形胶质细胞纯化培养,通过OGD模拟缺血缺氧诱导激活培养的原代星形胶质细胞。实验分为正常组,OGD组与OGD+缝隙通道阻断剂CBX组(OGD+CBX组)。采用2-NBDG检测星形胶质细胞在OGD后不同再灌时间(0h,12 h,24 h,48 h)的糖摄取能力以及免疫荧光方法观察缝隙连接蛋白43(Connexin 43,Cx43)数量与形态变化。结果星形胶质细胞经糖氧剥夺后,随着再灌时间延长糖摄取能力逐渐增加。再灌24 h OGD组糖摄取量为[(2.32±0,43)nmol/jigDNA],与对照组[(0.95±0.28)nmol/fjigDNA]相比糖摄取能力上调2.63倍(t=13.99,P=0.0024);再灌24 h时Cx43形成的缝隙斑点与0 h相比,缝隙连接数量增加2.5倍(t=11.34,P=0.003),斑点大小增加1.85倍(t=10.27,P=0.004)0与OGD组相比,OGD+CBX组再灌48 h星形胶质细胞糖摄取功能下降42%。结论OGD条件下星形胶质细胞Cx43缝隙连接增加糖摄取功能,可能为脑缺血临床治疗潜在靶点。Objective To explore the relationship between gap junction and glucose uptake of astrocytes under oxygen-glucose deprivation(OGD)and reperfusion.Methods Cerebral cortical astrocyte from 1 day newborn SD rats were undergone the primary culture.The ischemia cell model was established by OGD.This experiment were divided into control group,OGD group and OGD+CBX group.After different reperfusion time(0 h,12 h 24 h and 48 h),the glucose uptake of astrocyte was measured by 2-NBDG through flow cytometry analysis and connexin 43(Cx43)gap junction plaques was detected using immunofluorescene.Results Compared with the control group,the glucose uptake of astrocyte was up-regulated in・duced by OGD following different reperfusion time.The glucose uptake of OGD group was(2.32±0.43)nmol/jigDNA in 24 hours reperfusion time and was(0.95±0.28)nmol/jigDNA in control group.The upregulation was up to 2.63-fold increase(t=13.99,P=0.0024)in 24 hours after reperfusion.Compared with the control group,the Cx43 gap junction number was up to 2.5-fold increase(Z=11.34,P=0.003)and the size was 1.85-fold increase(t=10.27,P=0.004)in 24 h reperfusion.The glucose uptake of astrocyte after OGD was reduced by CBX and the decrease was 42%in 48 h after reperfusion.Conclusion Those results urges us consider the clinical treatment for interfering with Cx43 gap junction.

关 键 词：星形胶质细胞 缝隙连接蛋白43 缝隙连接 氧糖剥夺 糖摄取功能 

分 类 号：R74[医药卫生—神经病学与精神病学]