YAP/OPA1信号通路在丙泊酚减轻小鼠海马神经元氧糖剥夺-复氧复糖损伤中的作用  

Role of YAP/OPA1 signaling pathway in propofol-induced reduction of oxygen-glucose deprivation and restoration injury in hippocampal neurons

作　　者：王泽华 马晓燕 喻文立[5] Wang Zehua;Ma Xiaoyan;Yu Wenli(Department of Anesthesiology,Changzhi Medical College,Changzhi 046000,China;Department of Anesthesiology,Heping Hospital Affiliated to Changzhi Medical College,Changzhi 046000,China;Department of Anesthesiology,Changzhi People's Hospital,Changzhi 046000,China;Tianjin Medical University First Center Clinical College,Tianjin 300070,China;Department of Anesthesiology,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]长治医学院麻醉学系,长治046000 [2]长治医学院附属和平医院麻醉科,长治046000 [3]长治市人民医院麻醉科,长治046000 [4]天津医科大学一中心临床学院,天津300070 [5]天津市第一中心医院麻醉科,天津300192

出　　处：《中华麻醉学杂志》2023年第8期986-990,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(82072219)。

摘　　要：目的:评价Yes相关蛋白(YAP)/视神经萎缩蛋白1(OPA1)信号通路在丙泊酚减轻小鼠海马神经元氧糖剥夺-复氧复糖损伤中的作用。方法:取正常培养处于对数生长期的HT_(2)2小鼠海马神经元,采用随机数字表法分为4组(n=54):对照组(C组)、氧糖剥夺-复氧复糖组(OGD/R组)、丙泊酚组(P组)和丙泊酚+YAP沉默组(P+siRNA-YAP组)。采用无糖无氧孵育6 h、复氧复糖培养24 h的方法制备神经元氧糖剥夺-复氧复糖损伤模型。P组于复氧复糖即刻加入丙泊酚终浓度50μmol/L。P+siRNA-YAP组于模型制备前48 h转染siRNA-YAP。采用CCK-8法检测神经元活力,采用流式细胞术检测ROS含量和凋亡率,采用分光光度法检测MDA含量、SOD活性以及线粒体膜电位(MMP),采用荧光素荧光酶发光法检测线粒体ATP含量,采用免疫荧光法观察YAP核转位,Western blot法检测YAP、磷酸化YAP(p-YAP)以及OPA1的表达。结果:与C组比较,OGD/R组神经元活力降低,ROS、MDA含量和凋亡率升高,SOD活性、MMP和线粒体ATP含量降低,p-YAP表达上调,OPA1表达下调(P<0.05),核内YAP荧光强度减弱;与OGD/R组比较,P组神经元活力升高,ROS、MDA含量和凋亡率降低,SOD活性、MMP和线粒体ATP含量升高,p-YAP表达下调,OPA1表达上调(P<0.05),核内YAP荧光强度增强;与P组比较,P+siRNA-YAP组神经元活力降低,ROS、MDA含量和凋亡率升高,SOD活性、MMP和线粒体ATP含量降低,p-YAP、YAP和OPA1表达下调(P<0.05),核内YAP荧光强度减弱。结论:丙泊酚减轻小鼠海马神经元氧糖剥夺-复氧复糖损伤的机制可能与激活YAP/OPA1信号通路有关。Objective To evaluate the role of Yes-associated protein(YAP)/Optic atrophy-1(OPA1)signaling pathway in propofol-induced reduction of oxygen-glucose deprivation and restoration(OGD/R)injury in hippocampal neurons.Methods HT_(2)2 mouse hippocampal neurons at the logarithmic growth phase were divided into 4 groups(n=54 each)using a random number table method:control group(group C),group OGD/R,propofol group(group P)and propofol+YAP silencing group(group P+siRNA-YAP).The cells were subjected to O2-glucose deprivation for 6 h followed by restoration of O2-glucose supply for 24 h.In group P,propofol 50μmol/L was added immediately after restoration of O2-glucose supply.In P+siRNA-YAP group,siRNA-YAP was transfected at 48 h before model preparation.The viability of neurons was measured by CCK-8 assay,ROS content and apoptosis rate were measured by flow cytometry,the content of malondialdehyde(MDA),activity of superoxide dismutase(SOD)and mitochondrial membrane potential(MMP)were determined by spectrophotometry,the content of mitochondrial ATP was determined by fluorescein fluorescence method,the nuclear translocation of YAP was observed by immunofluorescence,and the expression of YAP,phosphorylated YAP(p-YAP)and OPA1 was detected by Western blot.Results Compared with group C,the viability of hippocampal neurons was significantly decreased,the contents of ROS and MDA and apoptosis rate were increased,the SOD activity,MMP and mitochondrial ATP content were decreased,the expression of p-YAP protein was up-regulated,OPA1 expression was down-regulated(P<0.05),and the fluorescence intensity of YAP in nucleus was weakened in group OGD/R.Compared with OGD/R group,the viability of neurons was significantly increased,the contents of ROS and MDA and apoptosis rate were decreased,the activity of SOD,MMP and content of mitochondrial ATP were increased,the expression of p-YAP protein was down-regulated,the expression of OPA1 protein was up-regulated(P<0.05),and the fluorescence intensity of YAP in nucleus was enhanced in P group.Com

关 键 词：二异丙酚 神经元 再灌注损伤 Yes相关蛋白 视神经萎缩蛋白1 

分 类 号：R614[医药卫生—麻醉学]