摘要
目的评价孤儿核受体Nur77在衣霉素(TM)诱导小鼠海马神经元损伤中的作用及其与内质网应激的关系。方法小鼠HT22细胞,采用随机数字表法分为4组(n=10):对照组(C组)、Nur77特异性激动剂Csn-B组(Csn-B组)、内质网应激诱导剂TM组(TM组)和TM+Csn-B组。C组细胞正常培养24 h;Csn-B组培养基中加入Csn-B(终浓度为10μg/ml)培养细胞24 h;TM组培养基中加入TM(终浓度为200 ng/ml)培养24 h诱导细胞内质网应激损伤;TM+Csn-B组培养基中加入Csn-B(终浓度为10μg/ml)预处理细胞15 min后,给予TM(终浓度为200 ng/ml)共同培养24 h。各组细胞处理结束后采用CCK-8试剂盒检测细胞活力,Western blot法检测内质网应激相关蛋白CCAAT/增强子结合蛋白同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)、凋亡相关蛋白Bcl-2、Bax、caspase-3和活化的caspase-3(cleaved-caspase-3)的表达。结果与C组比较,TM组细胞活力下降,CHOP、GRP78、Bax和cleaved-caspase-3表达上调,Bcl-2和caspase-3表达下调(P<0.05或0.01);与TM组比较,TM+Csn-B组细胞活力升高,CHOP、GRP78、Bax和cleaved-caspase-3表达下调,Bcl-2和caspase-3表达上调(P<0.05或0.01)。结论孤儿核受体Nur77参与了TM诱导小鼠海马神经元损伤的过程,与激活内质网应激有关。
Objective To evaluate the role of orphan nuclear receptor Nur77 in tunicamycin(TM)-induced injury to hippocampal neurons and the relationship with endoplasmic reticulum stress in mice.Methods Mouse HT22 cells were divided into 4 groups(n=10 each)using a random number table method:control group(C group),Nur77 specific agonist Csn-B group(Csn-B group),endoplasmic reticulum stress inducer TM group(TM group),and TM+Csn-B group.Cells in C group were cultured for 24 h under normal condition.In Csn-B group,Csn-B at a final concentration of 10μg/ml was added to the culture medium,and the cells were incubated for 24 h.In TM group,TM at a final concentration of 200 ng/ml was added to the culture medium and the cells were incubated for 24 h to induce cell endoplasmic reticulum stress injury.Cells in TM+Csn-B group were pretreated with Csn-B at a final concentration of 10μg/ml for 15 min,then TM at a final concentration of 200 ng/ml was added,and the cells were co-incubated for 24 h.The cell viability was examined by CCK-8 assay kit after treatment in each group.The expression of endoplasmic reticulum stress-related protein CCAAT/enhancer-binding protein homologous protein(CHOP),glucose regulated protein 78(GRP78)and apoptosis-associated protein Bcl-2,Bax,caspase-3 and cleaved-caspase-3 was detected by Western blot.Results Compared with C group,the cell viability was significantly decreased,and the expression of CHOP,GRP78,Bax and cleaved-caspase-3 was up-regulated,and the expression of Bcl-2 and caspase-3 was down-regulated in TM group(P<0.05 or 0.01).Compared with TM group,the cell viability was significantly increased,the expression of CHOP,GRP78,Bax and cleaved-caspase-3 was down-regulated,and the expression of Bcl-2 and caspase-3 was up-regulated in TM+Csn-B group(P<0.05 or 0.01).Conclusions Orphan nuclear receptor Nur77 is involved in TM-induced injury to hippocampal neurons,which is related to activation of the endoplasmic reticulum stress in mice.
作者
王小烨
刘天雅
周思琦
党静静
王志萍
Wang Xiaoye;Liu Tianya;Zhou Siqi;Dang Jingjing;Wang Zhiping(Jiangsu Province Key Laboratory of Anesthesiology,Xuzhou Medical University,Xuzhou 221004,China;Department of Anesthesiology,The Affiliated Hospital of Xuzhou Medical University,Xuzhou 221004,China;Department of Gastroenterology,Nanjing Drum Tower Hospital Clinical College of Jiangsu University,Nanjing 210008,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2023年第4期418-421,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(82270059)
江苏省卫健委医学科研重点项目(K2019003)
徐州市社会发展重点研发计划(KC21154)。
关键词
核受体亚家族4
A组
成员1
衣霉素
海马
神经元
内质网应激
Nuclear receptor subfamily 4,group a,member 1
Tunicamycin
Hippocampus
Neurons
Endoplasmic reticulum stress
