Conclusion
Inhibition of intercellular gap junction has protective effect against hypoxia/reoxygenation injury in astrocytes. 目的: 研究抑制细胞缝隙连接通讯功能对星形胶质细胞缺氧/复氧损伤的影响. 方法: 原代培养的新生SD大鼠大脑皮层星形胶质细胞分为正常对照组、空白对照组及细胞缝隙连接抑制剂18-α-甘草次酸和油酸酰胺组.荧光示踪法检测星形胶质细胞的缝隙连接通讯功能; MTT法检测抑制缝隙连接通讯功能对缺氧/复氧损伤星形胶质细胞存活率的影响; Annexin V/碘化丙啶双染流式细胞术及Hoechst 33258荧光染色法检测抑制缝隙连接通讯功能对缺氧/复氧损伤星形胶质细胞凋亡的影响. 结果: 与正常对照组相比, 空白对照组星形胶质细胞缝隙连接通讯功能明显增强( P＜0.01), 细胞存活率明显降低( P＜0.01), 细胞凋亡明显增加( P＜0.01);与空白对照组相比, 18-α-甘草次酸及油酸酰胺可显著降低星形胶质细胞缝隙连接通讯功能(均 P＜0.01), 减轻缺氧/复氧损伤引起的星形胶质细胞存活率的降低(均 P＜0.01), 且可减少缺氧/复氧损伤引起的星形胶质细胞凋亡(均 P＜0.01). 结论: 抑制细胞间缝隙连接通讯功能对星形胶质细胞缺氧/复氧损伤具有保护作用.
Methods
Primary cultured cerebral cortical astrocytes of neonate rats were divided into normal control group, hypoxia reoxygenation injury group and 18-α-glycyrrhetinic acid and oleamide (gap junctional intercellular channel inhibitors) group. The gap junction intercellular communication was determined by Parachute assay. The viability of astrocyes was detected by MTT assay. The apoptosis of astrocytes were detected with annexin V/PI and Hoechst 33258 staining.
Objective
To investigate the effects of inhibiting gap junctional intercellular communication on hypoxia/reoxygenation injury in astrocytes.
Results
Compared with the normal control group, the gap junctional function of astrocytes was increased significantly in ischemia/reperfusion group (P<0.01), the surviving fraction of astrocytes decreased significantly (P<0.01) and its cell apoptosis ratio increased significantly (P<0.01). Compared with the ischemia/reperfusion group, the gap junctional function of astrocytes in18-α-glycyrrhetinic acid and oleamide group decreased significantly (P<0.01), the viability of astrocytes increased significantly (P<0.01), while cell apoptosis decreased significantly (P<0.01).