GM-1预处理减轻布比卡因诱导的N2a细胞凋亡  

Effect of GM-1 pretreatment reducing the apoptosis after bupivacaine-induced in N2a cells

作　　者：刘丹[1] 罗茜 刘本铨 梁予洁 吉杰梅[1] 刘敬臣[1] LIU Dan;LUO Xi;LIU Benquan;LIANG Yujie;JI Jiemei;LIU Jingchen(Department of Anesthesiology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁市530021 [2]广东省佛山市第一人民医院麻醉科 [3]广西柳州市人民医院麻醉科

出　　处：《临床麻醉学杂志》2021年第3期286-289,共4页Journal of Clinical Anesthesiology

基　　金：国家自然科学基金资助项目(81660623),广西研究生教育创新计划项目(YCSW2020117)。

摘　　要：目的研究单唾液酸四己糖神经节苷脂(GM-1)预处理对布比卡因诱导N2a细胞凋亡后对c-Jun氨基末端激酶(JNK)、CCAAT/增强子结合蛋白同源蛋白(CHOP)表达的影响。方法体外培养小鼠神经母细胞瘤细胞N2a细胞株,取对数生长期N2a细胞,将细胞随机分为四组:对照组(C组),N2a细胞无任何药物处理;GM-1组(G组),N2a细胞中加入GM-15μmol/L处理24 h;布比卡因组(B组),N2a细胞中加入布比卡因900μmol/L处理36 h;GM-1预处理组(GB组),GM-15μmol/L预处理24h后,N2a细胞中加入布比卡因900μmol/L处理36 h。以倒置显微镜观察细胞形态学变化,采用TUNEL法检测细胞凋亡率,采用qRT-PCR法和Western blot法分别检测细胞内JNK和CHOP mRNA表达量及蛋白含量。结果与C组比较,B组和GB组细胞损伤形态明显加重,凋亡率明显升高,JNK和CHOP mRNA表达量及蛋白含量明显升高(P<0.05)。与B组比较,GB组细胞损伤形态明显减轻,凋亡率明显下降,JNK和CHOP mRNA表达量及蛋白含量明显降低(P<0.05)。结论GM-1预处理通过调控内质网应激凋亡途径中相关蛋白JNK和CHOP的表达,从而减少布比卡因诱导的细胞凋亡,减轻布比卡因引起的神经毒性。Objective To measure the effect of expressions of c-JNK N-terminal kinase(JNK),CCAAT/enhancer binding protein homologous protein(CHOP)after monosialoganglioside(GM-1)pretreatment on bupivacaine-induced apoptosis in N2a cells.Methods Mice nerve N2a cells cultured in vitro.Growing N2a cells were randomly divided into four groups:the control group(group C),N2a cells were treated without any drug;GM-1 group(group G),N2a cells were cultured with 5μmol/L GM-1 for 24 hours;bupivacaine group(group B),N2a cells were treated with 900μmol/L bupivacaine for 36 hours;GM-1 pretreatment group(group GB),N2a cells were pretreated with GM-1 for 24 hours and then added 900μmol/L bupivacaine for 36 hours.Light Microscope was detected to observe the morphological changes of the cells of N2a.TUNEL assay was measured the apoptosis rate of N2a cells.Real-time polymerase chainreaction and Western blotting evaluated the expressions of JNK and CHOP.Each expreiment was repeated three times.Results Compared with group C,N2a cells of groups B and GB were seriously damaged under the light microscope and the apoptosis rate was incresead,the mRNA and protein expressions of JNK and CHOP were significantly increased(P<0.05).Compared with group B,N2a cells of group GB had less cell damage morphology,and the apoptosis rate was reduced,mRNA and protein expressions of JNK and CHOP were significantly reduced(P<0.05).Conclusion GM-1 pretreatment regulated the expression of JNK and CHOP in the apoptosis pathway of endoplasmic reticulum stress,thereby reducing bupivacaine-induced cell apoptosis and had a certain protective effect on bupivacaine-induced neurotoxicity in N2a cells.

关 键 词：布比卡因 神经毒性 单唾液酸四己糖神经节苷脂 C-JUN氨基末端激酶 CCAAT/增强子结合蛋白同源蛋白 N2A细胞 细胞凋亡 

分 类 号：R965[医药卫生—药理学]