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1d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,12h,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/μ gDNA],与对照组[(0.95±0.28) nmol/μgDNA]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0h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).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.

abstractsObjective 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 induced by OGD following different reperfusion time.The glucose uptake of OGD group was (2.32 ± 0.43)nmol/μgDNA in 24 hours reperfusion time and was (0.95±0.28)nmol/μgDNA in control group.The upregtulation 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(t=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.