Bax、Bcl-2在培养新生鼠皮层神经元中的表达及L-NAME的干预作用

doi:10.13418/j.issn.1001-165x.2014.05.020

中国临床解剖学杂志 ›› 2014, Vol. 32 ›› Issue (5): 595-598.doi: 10.13418/j.issn.1001-165x.2014.05.020

Bax、Bcl-2在培养新生鼠皮层神经元中的表达及L-NAME的干预作用

宋海岩，　高志涛，　邓晓慧，　连辉，　任铭新，　付升旗

新乡医学院河南省医用组织再生重点实验室，河南新乡 453003

收稿日期:2013-10-24 出版日期:2014-09-25 发布日期:2014-10-14
通讯作者: 任铭新,讲师，Tel:（0373）3029051，E-ail:hnlyrmx@163.com E-mail:shy80825@163.com
作者简介:宋海岩（1980-），女，河南人，硕士，讲师，主要从事神经损伤研究，Tel: 0373-3029051
基金资助:
2011年新乡医学院重点研究领域招标课题（ZD2011-14)

The expression of Bax and Bcl-2 in cultured neonatal rat cortical neurons and the role of L-NAME intervention

SONG Hai-yan, GAO Zhi-tao, DENG Xiao-hui, LIAN Hui, REN Ming-xin, FU Sheng-qi

Xinxiang Medical University；Key Laboratory for Medical Tissue Regeneration of Henan Province, Xinxiang 453003, China

Received:2013-10-24 Online:2014-09-25 Published:2014-10-14

摘要/Abstract

摘要：

目的　探讨Bax与Bcl-2在体外培养的皮层神经元的表达及非选择性一氧化氮合酶抑制剂NG-硝基-L-精氨酸甲酯（L-NAME）的干预作用。方法　原代培养SD新生大鼠皮层神经元，并采用neuron specific enolase (NSE)免疫荧光方法进行细胞纯度鉴定。神经元随机分为正常组、N-甲基-D-天门冬氨酸（NMDA）组、NG-Nitro-L-arginine Methyl Ester（L-NAME）组，分别采用ELISA检测各组细胞中一氧化氮合酶（nitric oxide synthase, NOS）的表达，采用RT-PCR、Western blot分别检测各组细胞中Bax、Bcl-2 mRNA及蛋白的表达。结果　NSE免疫荧光大多数细胞为NSE阳性细胞，神经元纯度高达90%；L-NAME组中NOS及Bax的表达均高于正常组而低于NMDA组， Bcl-2的表达低于正常组而高于NMDA组。结论　L-NAME对NMDA诱导的神经细胞凋亡起明显的保护作用，其可能是通过上调Bcl-2的表达、下调Bax的表达发挥作用。

关键词: 一氧化氮合酶, Bax, Bcl-2

Abstract:

Objective　To investigate the expression of Bcl-2 and Bax in cultured cortical neurons after NMDA injury and intervention effect of L-NAME.　Methods 　The primary cortical neurons were identified by polyclonal antibody against NSE. The neurons were randomly divided into 3 groups: control group, NMDA-injured group, L-NAME pretreatment group. ELISA was employed to explore the expression of NOS, and the expression of Bcl-2 and Bax was detected by RT-PCR and Western blot. 　Results 　Immunofluorescence test showed that the most cells were NSE positive staining cells. NOS in L-NAME group were higher than that in control group, but lower than that in NMDA group; the expression of Bcl-2 mRNA and protein in L-NAME group were lower than in control group, but higher than that in NMDA group. The expression of Bax was in contrast with Bcl-2.　Conclusion　L-NAME plays an important protective role in neuronal apoptosis induced by NMDA, the possible mechanism of which may be related with up-regulation of Bcl-2 expression and down-regulation of Bax expression.

Key words: NOS, Bax, Bcl-2

中图分类号:

R363

宋海岩, 高志涛, 邓晓慧, 连辉, 任铭新, 付升旗. Bax、Bcl-2在培养新生鼠皮层神经元中的表达及L-NAME的干预作用[J]. 中国临床解剖学杂志, 2014, 32(5): 595-598.

SONG Hai-Yan, GAO Zhi-Chao, DENG Xiao-Hui, LIAN Hui, LIN Ming-Xin, FU Sheng-Qi. The expression of Bax and Bcl-2 in cultured neonatal rat cortical neurons and the role of L-NAME intervention[J]. Chinese Journal of Clinical Anatomy, 2014, 32(5): 595-598.

参考文献

[1] 刘伟困，翟丽，张浩，等. 一氧化氮调节细胞凋亡的作用机制
[J]. 生命的化学，2009，28(1)：45-48．

[2] 胡文忠，王晓强. 一氧化氮与一氧化氮合酶抑制剂对脑缺血后海马神经元的保护作用
[J]. 中国实用神经疾病杂志，2011，14（3）：8-10.

[3] 宋海岩，王志勇，李娜娜. 新生大鼠大脑皮质神经元的原代培养及其鉴定
[J]. 实用儿科临床杂志, 2012, 27(2):129-131.

[4] 戴晓春，周希，黄晓燕，等.转录因子Pax-8基因干扰后线粒体功能对心肌细胞凋亡的影响
[J].中华心血管病杂志，2013，41(1):54-59.

[5] Nicoll RA, Malenka RC. Expression mechanisms underlying NMDA receptor-dependent long-term potentiation
[J]. Ann N Y Acad Sci, 1999,868: 515-525.

[6] Malenka RC, Nicoll RA. NMDA-receptor-dependent synaptic plasticity: multiple forms and mechanisms
[J]. Trends Neurosci, 1993, 16(12):521-527.

[7] Bliss TV, Collingridge GL. A synaptic model of memory:long-term potentiation in the hippocampus
[J]. Nature, 1993, 361(6407):31-39.

[8] Brennan AM, Won Suh S, Joon Won S, et al. NADPH oxidase is the primary source of superoxide induced by NMDA receptor activation
[J]. Nat Neurosci, 2009, 12(7):857–863.

[9] Chen X, Liu J, Gu X, et al. Salidroside attenuates glutamate-induced apoptotic cell death in primary cultured hippocampal neurons of rats
[J]. Brain Res, 2008, 31 (1238):189-198.

[10]金宏波, 徐洪伟, 刘军, 等.一氧化氮合酶抑制剂L-NNA对福尔马林大鼠脑nNOS、iNOS表达的影响
[J].哈尔滨医科大学报,2004,38(5),407-409.

[11]Ortiz-Ortiz MA, Moran JM, Gonzalez-Polo RA, et al. Nitric oxide-mediated toxicity in paraquat-exposed SH-SY5Y cells: a protective role of 7-nitroindazole
[J]. Neurotox Rex, 2009, 16(2):160-173.

[12]Iriyama T, Kamei Y, Kozuma S, et al. Bax-inhibiting peptide protects glutamate-induced cerebellar granule cell death by blocking Bax translocation
[J]. Neurosci Lett, 2009, 451(1):11-15.

[13]Chen F, Chen Y, Kang X, et al. Anti-apoptotic function and mechanism of ginseng saponins in Rattus pancreatic β-cells
[J]. Biol Pharm Bull, 2012, 35(9):1568-1573.

[14]高琴，胡俊峰，于影，等. 一氧化氮在乙醇后处理心肌保护中的作用
[J]. 中国应用生理学杂志 2012, 28(1): 9-13.

[15]Gross A, Jockel J, Wei MC, et al. Enforcec cimerization of BAX results in its translocation, mitochoncrial cysfunction and apoptosis
[J]. EMBO J, 1998, 17 (14):3878-3885.

[16]葛宇松,尹琳,滕伟禹,等.亲环素A对Aβ25-35诱导PC12细胞凋亡的影响及机制研究
[J].中华神经医学杂志，2011，10(6):582-586.

[17] Mao W, Yi X, Qin J, et al. CXCL12 inhibits cortical neuron apoptosis by increasing the ratio of Bcl-2/Bax after traumatic brain injury
[J]. Int J Neurosci, 2014, 124(4):281-290.

Bax、Bcl-2在培养新生鼠皮层神经元中的表达及L-NAME的干预作用

The expression of Bax and Bcl-2 in cultured neonatal rat cortical neurons and the role of L-NAME intervention

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

Metrics

本文评价

推荐阅读 0

[1]	胡晓芳，邱仁杰，余磊，刘茂生，杨琳，王国保. 纳米载体SWCNT投递Bcl-2 siRNA促进U251细胞凋亡[J]. 中国临床解剖学杂志, 2017, 35(5): 521-525.
[2]	汤银娟,王建钧,王祥海,潘梦婕,陈卓,肖玲珑,刘嘉华,蔡维君. 茶多酚对慢性酒精中毒肝损伤影响的实验研究[J]. 中国临床解剖学杂志, 2016, 34(2): 198-201.
[3]	汤银娟, 王建钧, 王祥海, 文锦坤, 潘梦婕, 钱长晖, 康伟祥, 陈卓. 茶多酚预防慢性酒精中毒对精子损伤的作用及机制研究[J]. 中国临床解剖学杂志, 2014, 32(5): 604-608.
[4]	方志启, 吴刚, 王贺彬, 陈晓宇. 凋亡相关基因p53、bcl-2、bax在前列腺组织中的相关性[J]. 中国临床解剖学杂志, 2013, 31(5): 560-563.
[5]	宋海岩, 李娜娜, 付升旗, 任铭新. Bax、NOS在NMDA诱导的兴奋性神经毒中的表达变化[J]. 中国临床解剖学杂志, 2012, 30(6): 675-677.