丙泊酚对小鼠少突神经胶质细胞缺血缺氧再灌注损伤的保护  

Protection of propofol against ischemia-hypoxia reperfusion injury of mouse glial cells

作　　者：王昆锋 丁洁 王洪强 WANG Kun-feng;DING Jie;WANG Hong-qiang(The Fifth Affiliated Hospital of Shenzhen University(Shenzhen Baoan Central Hospital)Department of Anesthesiology 518000 Shenzhen,Guangdong Province)

机构地区：[1]深圳大学第五附属医院/深圳宝安中心医院麻醉科,广东深圳518000

出　　处：《解剖学研究》2021年第3期236-240,共5页Anatomy Research

摘　　要：目的探讨丙泊酚对小鼠少突神经胶质细胞缺血缺氧/再灌注损伤中的保护作用及可能机制。方法将处于对数生长期的小鼠少突神经胶质细胞,体外构建缺血缺氧/再灌注损伤细胞模型。随机将细胞分为为4组(每组10个样本)、常规培养组(N组)、丙泊酚常规组(P组)、缺血缺氧再灌注组(H组)和缺血缺氧再灌注+丙泊酚组(HP组)。P组和HP组在造模开始时加入10μmol/L的丙泊酚;N组和P组在培养箱常规培养27 h;H组和HP组细胞缺血缺氧培养3 h,再复氧复血继续培养24 h。之后用倒置显微镜观察各组细胞形态学变化,采用CCK-8法检测各组细胞活力,流式法检测各组细胞的凋亡。Western blot检测各组caspase-3和Bcl-2蛋白的表达。结果倒置显微镜观察HP组与H组比较,细胞肿胀变形程度较轻,未见细胞核明显固缩和大量细胞发生融合凋亡现象,上清液仅可见少量细胞碎片及悬浮凋亡细胞。CCK-8检测结果,HP组与H相比,A值上调,差异有统计学意义(P<0.01)。流式检测细胞凋亡结果,HP组与H相比,细胞存活率增高,凋亡细胞和死亡细胞减少(P<0.01)。Western blot蛋白检测示:HP组与H相比,caspase-3蛋白表达量明显降低(P<0.01);HP组与H相比,Bcl-2蛋白表达量明显升高(P<0.01)。结论丙泊酚能抑制caspase-3蛋白表达,提高Bcl-2蛋白表达,减轻缺血缺氧再灌注损伤导致的少突神经胶质细胞的凋亡现象,增强损伤细胞的活性。Objective To investigate the protective effect and possible mechanism of propofol on oligodendrocytes in mice from ischemia-hypoxia/reperfusion injury.Methods The mouse oligodendrocytes in the logarithmic growth phase were used to construct a cell model of hypoxia/reperfusion injury in vitro.The cells were randomly divided into 4 groups:conventional culture group(N group),conventional propofol group(P group),hypoxia-ischemia reperfusion group(H group)and hypoxia-ischemia re perfusion+propofol group(HP group).P group and HP group 10μmol/L propofol was added at the beginning of model building.N group and P group were routinely cultured in an incubator for 27 hours.The cells in the H group and HP group were cultured for 3 hours after ischemia and hypoxia,and then reoxygenated and re-blooded for 24 hours.After that,an inverted microscope was used to observe the morphological changes of the cells in each group,the cell viability of each group was detected by the CCK-8 method,and the apoptosis of the cells in each group was detected by the flow method.Western blotting was performed to detect the expression of caspase-3 and Bcl-2 proteins in each group.Results Compared with the H group,the HP group was observed with an inverted microscope.The degree of cell swelling and deformation was lighter.There was no obvious pyknosis of the nucleus and a large number of cells fused and apoptosis.Only a small amount of cell debris and suspended apoptotic cells were seen in the supernatant.As a result of CCK-8 test,the OD value of HP group was increased compared with H group,and the difference was statistically significant(P<0.01).As a result of flow cytometry,compared with H group,the cell survival rate of HP group was increased,and apoptotic cells and dead cells were decreased(P<0.01).Westernblot protein detection showed that the expression of caspase-3 protein in HP group was significantly lower than that in H group(P<0.01).Compared with H group,the expression of Bcl-2 protein in the HP group was significantly increased(P<0.01).

关 键 词：丙泊酚 少突神经胶质细胞 缺血缺氧/再灌注 凋亡 

分 类 号：R614[医药卫生—麻醉学]