泻根醇酸对NMDA诱导的PC12细胞损伤的保护作用研究  

Bryonolic Acid Protects Against NMDA-Induced Neurotoxicity in PC12 Cells

作　　者：张胜男[1] 李煌[1] 张玉琴[1] 徐伟[1] 陈立典[1] 黄枚[1] 褚克丹[1] 阙金花 

机构地区：[1]福建中医药大学药学院,福州450122

出　　处：《天然产物研究与开发》2015年第1期139-142,162,共5页Natural Product Research and Development

基　　金：福建省科技厅重点项目(2012Y0041);福建省教育厅项目(JA12176);国家级大学生创新创业训练计划项目(201410393008);校管课题(X2013015)

摘　　要：钙离子内流、Ca2+/钙调蛋白(Ca M)依赖的蛋白激酶Ⅱ(CaMKⅡ)和c AMP应答元件结合蛋白(CREB)的磷酸化是NMDA诱导细胞凋亡的重要过程。本实验探讨了泻根醇酸(BA)对N-甲基-D-天冬氨酸(NMDA)诱导的PC12细胞损伤的保护作用及可能机制。MTT法测定细胞活力、测定LDH释放率、Fura-2/AM荧光标记法测定细胞内钙离子浓度,Western blot法测定CaMKⅡ、p-CaMKⅡ、CREB、p-CREB、Bax、Bcl-2蛋白表达。结果显示,与模型组相比,BA给药组细胞活力显著升高,LDH释放减少,[Ca2+]i降低,p-CREB,Bcl-2表达上调,Bax表达下调。BA能够通过Ca2+-CaMKⅡ-CREB信号通路保护NMDA诱导的PC12细胞损伤,进而有望成为脑缺血疾病的神经保护药物。Calcium overload is considered as one of the mechanisms of cerebral ischemia. Ca2 ＋influx and Ca2 ＋/ calmodulin-dependent protein kinase Ⅱ（CaMKⅡ） and c AMP response element-binding protein（CREB） phosphorylation are considered to be involved in N-Methyl-D-aspartate（NMDA）-induced apoptosis process. In this study,we investigated the neuroprotective effects of bryonolic acid（BA） in the NMDA-induced rat＇s adrenal pheochromocytoma cell line（PC12 cells） and the potential mechanism. NMDA induced cytotoxicity in PC12 cells was accompanied by cell viability decrease and lactate dehydrogenase（LDH） release,as well as Ca2 ＋influx,Bax up-regulation,p-CREB and Bcl-2 downregulation. The results showed that pretreatment with BA significantly attenuated cells viability decrease and LDH release,as well as Ca2 ＋influx,Bax generation p-CREB and Bcl-2 protein increase. All these results indicated that BA protected PC12 cells against NMDA-induced apoptosis by inhibiting Ca2 ＋influx and regulating genes expression in Ca2 ＋-CaMKⅡ-CREB signal pathway. Therefore,the present study supported the notion that BA may be a promising neuroprotective agent for the treatment of cerebral ischemia disease.

关 键 词：泻根醇酸 脑缺血 PC12细胞 CA2+ Bcl-2 Bax p-CaMKⅡ P-CREB 

分 类 号：R285.5[医药卫生—中药学]