丙泊酚减轻人胶质细胞瘤细胞糖剥夺损伤时miR-29a与PUMA的关系  

Relationship between microRNA-29a and PUMA in propofol-induced reduction of glucose deprivation-induced injury to human giloma cells

作　　者：王云霞[1] 谭泽霞 张鑫磊 周相君 陆瑜[1] Wang Yunxia;Tan Zexia;Zhang Xinlei;Zhou Xiangjun;Lu Yu(Department of Anesthesiology,Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China)

机构地区：[1]首都医科大学附属北京天坛医院麻醉科,北京100070

出　　处：《中华麻醉学杂志》2024年第12期1495-1498,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81201017);北京市卫生系统高层次卫生技术人才培养计划-学科骨干(2015-3-039)。

摘　　要：目的评价丙泊酚减轻人胶质细胞瘤细胞糖剥夺损伤时微小RNA-29a(miR-29a)与p53上调凋亡调控因子(PUMA)的关系。方法体外培养人胶质细胞瘤U87细胞至对数生长期,采用随机数字表法分为6组(n=24):对照组(C组)、糖剥夺组(GD组)、丙泊酚+糖剥夺组(P+GD组)、miR-29a抑制剂组(I组)、miR-29a抑制剂组+糖剥夺组(I+GD组)、miR-29a抑制剂+丙泊酚+糖剥夺组(I+P+GD组)。C组在正常条件下培养;GD组更换为无糖DMEM培养基,培养12 h;P+GD组加入丙泊酚终浓度为10μmol/L孵育12 h,弃去培养基,更换为无糖DMEM培养基,培养12 h。I组、I+GD组和I+P+GD组转染miR-29a抑制剂孵育48 h,后续处理同P+GD组。测定细胞存活率、线粒体膜电位和ROS水平,采用qRT-PCR法检测miR-29a表达,Western blot法检测PUMA表达。结果与C组比较,GD组和I组细胞存活率和线粒体膜电位降低,ROS水平升高,细胞miR-29a表达下调,PUMA表达上调(P<0.05);与GD组比较,P+GD组细胞存活率和线粒体膜电位升高,ROS水平降低,miR-29a表达上调,PUMA表达下调,I+GD组细胞存活率和线粒体膜电位降低,ROS水平升高,miR-29a表达下调,PUMA表达上调(P<0.05);与P+GD组比较,I+P+GD组细胞存活率和线粒体膜电位降低,ROS水平升高,miR-29a表达下调,PUMA表达上调(P<0.05)。结论丙泊酚减轻人胶质细胞瘤细胞糖剥夺损伤的机制与上调miR-29a表达,下调PUMA表达有关。Objective To evaluate the relationship between microRNA-29a(miR-29a)and p53 up-regulated modulator of apoptosis(PUMA)in propofol-induced reduction of glucose deprivation(GD)-induced injury to human glioma cells.MethodsHuman glioma U87 cells were cultured in vitro to the logarithmic growth phase.Cells were then divided into 6 groups(n=24 each)by the random number table method:control group(group C),group GD,propofol+GD group(group P+GD),miR-29a inhibitor group(group I),miR-29a inhibitor+GD group(group I+GD)and miR-29a inhibitor+propofol+GD group(group I+P+GD).Cells were cultured in normal condition in group C.The culture medium was changed to glucose-free DMEM solution,and the cells were cultured for 12 h in group GD.In group P+GD,cells were incubated with propofol 10μmol/L for 12 h and then incubated in glucose-free DMEM solution for 12 h.In group I,group I+GD and group I+P+GD,miR-29a inhibitor was transfected into cells using lipofectamine transfection kit,and then the cells were cultured for 48 h,and the other treatments were similar to those previously described in group P+GD.The cell survival rate,mitochondrial membrane potential and level of reactive oxygen species(ROS)were determined.The expression of miR-29a was detected by quantitative real-time polymerase chain reaction,and the expression PUMA was detected by Western blot.ResultsCompared with group C,the cell survival rate and mitochondrial membrane potential were significantly decreased,the level of ROS was increased,the expression of miR-29a was down-regulated,and the expression of PUMA was up-regulated in group GD and group I(P<0.05).Compared with group GD,the cell survival rate and mitochondrial membrane potential were significantly increased,the level of ROS was decreased,the expression of miR-29a was up-regulated,and the expression of PUMA was down-regulated in group P+GD,and the cell survival rate and mitochondrial membrane potential were significantly decreased,the level of ROS was increased,the expression of miR-29a was down-regulated,and the e

关 键 词：二异泊酚 脑缺血 微RNAS 凋亡调节蛋白质类 

分 类 号：R73[医药卫生—肿瘤]