BI-D1870对N2a细胞毒作用  

Cytotoxic Effect of BI-D1870 in N2a Cells

作　　者：易妍 商亚珍[1] YI Yan;SHANG Ya-zhen(Institute of Traditional Chinese Medicine,Chengde Medical College,Hebei Province Key Research Office of Traditional Chinese Medicine Against Dementia,Hebei Province Key Laboratory of Traditional Chinese Medicine Research and Development,Hebei Key Laboratory of Nerve Injury and Repair,Chengde,067000,China)

机构地区：[1]承德医学院中药研究所,河北省中医药抗痴呆重点研究室,河北省中药研究与开发重点实验室,河北省神经损伤与修复重点实验室,承德067000

出　　处：《神经药理学报》2023年第3期1-9,共9页Acta Neuropharmacologica

基　　金：河北省自然基金资助项目(No.H2019406063);河北省中医药管理局资助项目(No.05027,No.2014062);河北省教育厅重点项目(No.ZD20131022,No.ZD2019057);河北省中药学重点学科建设项目;河北省中医药中药药理重点学科建设项目(冀中医药函[2021]12号);承德医学院科技创新团队项目(No.[2020]50)。

摘　　要：目的:探讨RSK抑制剂BI-D1870对N2a细胞毒作用,确定产生细胞毒作用的浓度和时间。方法:N2a细胞体外培养,免疫荧光法鉴定神经细胞特异性蛋白神经源性分化因子1(neurogenic differentiation 1,NeuroD1)、神经元核抗原(neuronal nuclei antigen,NeuN)和双皮质素(doublecortex,DCX)。N2a细胞用10~80μmol·L^(-1)BI-D1870,分别作用4~24 h。倒置显微镜观察N2a细胞形态,CCK-8法检测细胞存活率,丙酮酸还原法检测细胞培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)释放量。结果:免疫荧光可检测到N2a细胞有NeuroD1、NeuN和DCX阳性表达。与对照组比较,随着BI-D1870作用浓度增加和作用时间延长,N2a细胞受损程度逐渐增加,细胞形态发生异常改变,细胞存活率逐渐降低(P<0.05),培养液中LDH先升高后降低(P<0.05)。40μmol·L^(-1)作用12 h条件下N2a细胞肿胀明显,折光率降低,细胞存活率在60%左右,LDH释放量增长率最高。结论:N2a细胞具有神经细胞特性,BI-D1870对N2a细胞有细胞毒作用,产生细胞毒作用的最适条件为40μmol·L^(-1)作用12 h。Objective:To investigate the cytotoxic effect of RSK inhibitor BI-D1870 in N2a and determine the concentration and time of cytotoxic effect.Methods:N2a cells were cultured in vitro,and the specific proteins NeuroDl,NeuN and DCX were identified by immunofluorescence.N2a cells were exposed to a series concentration of 10μmol.L^(-1)to 80μmol-LI BI-D1870 and a series duration of 4 h to 24 h.The morphology of N2a cell was observed with the inverted microscope,and the cell viability was measured with CCK-8 method,and lactate dehydrogenase(LDH)release of cell culture medium was assayed by pyruvate reduction method.Results:NeuroD1,NeuN and DCX were detected in N2a cells by immunofluorescence.Compared with the DMSO solvent control group,with the increase of BI-D1870 concentration and the exposure prolongation of BI-D1870,the damage degree of N2a cells gradually increased.The cell morphology changed abnormally,the cell viability decreased gradually(P<0.05)and LDH level in culture medium went up and then went down(P<0.05).Under the treatment of 40μmol.L-'for 12 h,the swelling of N2a cells was obvious,the refraction was reduced,the cell viability was about 60%,and the LDH release in culture medium was the highest.Conclusion:N2a cells have the characteristics of neuron.BI-D1870 has cytotoxic effect for N2a cells and the optimal cytotoxic condition is 40μmol.L1 for 12 h.

关 键 词：BI-D1870 N2A细胞 乳酸脱氢酶 细胞毒作用 

分 类 号：R964[医药卫生—药理学]