hsacirc0081596在人神经细胞氧糖剥夺/复糖复氧损伤中的作用  

Role of hsa_circ_0081596 in oxygen-glucose deprivation and restoration injury to human neurons

作　　者：黄超 刘冰[2] 袁阳 王培 陈怀龙[2] 王明山[2] Huang Chao;Liu Bing;Yuan Yang;Wang Pei;Chen Huailong;Wang Mingshan(Graduate School of Dalian Medical University,Dalian 116044,China;Department of Anesthesiology,Qingdao Municipal Hospital Affiliated to Qingdao University,Qingdao 266071,China)

机构地区：[1]大连医科大学研究生院,116044 [2]青岛大学附属青岛市市立医院麻醉科,266071

出　　处：《中华麻醉学杂志》2021年第3期363-366,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价hsacirc0081596在人神经细胞氧糖剥夺/复糖复氧(OGD/R)损伤中的作用。方法培养人神经母细胞瘤细胞,采用随机数字表法将细胞(5代以内)分为4组(n=20):对照组(C组)、OGD/R组(O组)、OGD/R+小干扰RNA(siRNA)组(S组)、OGD/R+siRNA阴性对照组(I组)。C组细胞在37℃、5%CO_(2)正常条件下培养,O组细胞铺于6孔板或96孔板待完全贴壁,氧糖剥夺4 h后复糖复氧24 h制备OGD/R损伤模型。S组和I组分别转染hsacirc0081596 siRNA及其阴性对照,72 h后建立OGD/R模型。采用qRT-PCR法测定hsacirc0081596和线粒体分裂蛋白1(Fis1)mRNA的表达水平,采用Western blot发测定Fis1的表达水平,采用CCK-8法确定细胞存活率,采用流式细胞仪检测细胞凋亡率。结果与C组比较,O组hsacirc0081596、Fis1及其mRNA表达上调,细胞存活率降低,凋亡率升高(P<0.05);与O组比较,S组hsacirc0081596、Fis1表达下调,细胞存活率升高,凋亡率降低,I组hsacirc0081596、Fis1表达上调,细胞存活率降低,凋亡率升高(P>0.05)。结论hsacirc0081596可通过上调Fis1表达参与人神经细胞OGD/R损伤的病理生理机制。Objective To evaluate the role of hsa_circ_0081596 in oxygen-glucose deprivation and restoration(OGD/R)injury to human neurons.Methods SK-N-SH cells were cultured and the cells within 5 generations were divided into 4 groups(n=20 each)using a random number table method:control group(group C),OGD/R group(group O),OGD/R+siRNA group(group S)and OGD/R+siRNA negative control group(group I).The cells in C group were cultured under normal conditions of 37℃and 5%CO_(2).The cells in group O were placed in 6-or 96-well plates until they were completely attached to the wall,and then subjected to oxygen-glucose deprivation for 4 h,followed by restoration of oxygen-glucose for 24 h.In group S and group I,the cells were transfected with hsa_circ_0081596 siRNA and its negative control,respectively,and 72 h later OGD/R model was established.The expression of hsa_circ_0081596 and mitochondrial fission protein 1(Fis1)mRNA was detected using quantitative real-time polymerase chain reaction.The expression of Fis1 was determined by Western blot,the cell survival rate was determined by CCK-8 assay and the apoptosis rate was determined by flow cytometry.Results Compared with group C,the expression of hsa_circ_0081596,Fis1 and its mRNA was significantly up-regulated,the cell survival rate was decreased,and the apoptosis rate was increased in group O(P<0.05).Compared with group O,the expression of hsa_circ_0081596 and Fis1 was significantly down-regulated,the cell survival rate was increased and the apoptosis rate was decreased in group S,and the expression of hsa_circ_0081596 and Fis1 was significantly up-regulated,the cell survival rate was decreased and the apoptosis rate was increased in group I(P>0.05).Conclusion hsa_circ_0081596 is involved in the pathophysiological mechanism of OGD/R through up-regulating the expression of Fis1 in human neurons.

关 键 词：RNA 环状 脑 再灌注损伤 

分 类 号：R743.3[医药卫生—神经病学与精神病学]