川芎嗪对缺氧缺糖损伤大脑皮层神经元的保护作用及初步机制研究  

Preliminary study of neuroprotection and the mechanism of TMP on damage of cortical neurons by oxygen-glucose deprivation

作　　者：周新华[1] 王亮[2] 张在军[1] 于沛[1] 

机构地区：[1]暨南大学药学院新药研究所,广东广州510632 [2]澳门大学中华医药研究院中药质量研究国家重点实验室,澳门999078

出　　处：《暨南大学学报（自然科学与医学版）》2014年第6期532-537,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省医学科研基金项目(A2013353);广州市科技计划项目(132000726);暨南大学科研培育与创新基金项目(21611321;21613331)

摘　　要：目的:研究川芎嗪(TMP)对缺氧缺糖(OGD)损伤大脑皮层神经元的保护作用,探讨TMP神经保护作用的可能机制.方法:建立体外原代大脑皮层神经元OGD模型,利用MTT法和LDH试剂盒分别检测细胞存活率及细胞毒性;Hoechst染色法检测细胞凋亡;流式细胞仪检测细胞内ROS的含量;实时-PCR检测细胞内线粒体生物合成相关mRAN(PGC-1α,TFAM)的表达量.结果:经OGD损伤4 h后,细胞活力明显下降,细胞释放的LDH含量升高;细胞凋亡形态学特征明显,细胞凋亡率增加;细胞内ROS含量增加,同时PGC-1α和TFAM mRNA的表达量明显减少.TMP预保护后,可以明显提高OGD损伤后细胞的存活率,降低LDH的释放,改善细胞核形态变化,并且减少细胞凋亡率,抑制细胞内ROS的产生,提高PGC-1α和TFAM的表达.结论:TMP对OGD损伤的神经元具有神经保护作用,其机制可能与抗氧化,抗凋亡和增加线粒体生物合成相关因子(PGC-1α,TFAM)的表达有关.Aim:To investigate the effect of TMP in preventing primary cortical neurons (CNs)from OGD-induced cell damage and to elucidate the possible mechanisms of TMP.Methods:Primary cortical neurons were exposed to oxygen-glucose deprivation as an in vitro model of ischemic stroke.Cell viability was measured by using MTT assay and the cytotoxicity was assessed by determination of lactate dehydro-genase (LDH)that was released immediately after OGD.The apoptosis rate of neurons was detected by Hoechst 33342 staining.The level of intracellular reactive oxygen species(ROS)level was detected by DCFH-DA probe.Mitochondrial biogenesis-related factors(PGC-1αand TFAM)were measured using real time-PCR assay.Results:TMP significantly increased OGD-induced cell viability and attenuated neuro-toxicity,mitigated cell apoptosis,and alleviated OGD-induced ROS production in CNs.TMP also in-creased the mRNA transcription of PGC-1αand TFAM.Conclusion:TMP significantly protects CNs from OGD-induced cell damage.The TMP’s protective effect might be mediated by its ROS-scavenging activi-ty and increase of the transcription of mitochondrial biogenesis-related factors including PGC-1αand TFAM.

关 键 词：川芎嗪 原代大脑皮层神经元 缺氧缺糖模型 活性氧自由基 线粒体生物合成相关因子 

分 类 号：R285[医药卫生—中药学]