丹参川芎嗪注射液对Aβ损伤的PC12细胞保护作用及机制研究  

Protective effects and mechanism of salvia and TMP injection on Aβ-induced injury in PC12 cells

作　　者：严钢莉[1] 李朝武[1] 聂海岭[1] 黎逢光[1] 成勇[1] 毛高峰[1] 方煌[1] 魏海燕[1] 谢军[1] 

机构地区：[1]解放军161医院神经内科,武汉430012

出　　处：《卒中与神经疾病》2016年第1期15-20,共6页Stroke and Nervous Diseases

基　　金：湖北省卫计委科研基金资助项目(批准号WJ2015MB130)

摘　　要：目的探讨丹参川芎嗪注射液对Aβ损伤的PC12细胞可能的保护作用及机制。方法将PC12细胞分为5组:空白对照组(未加任何处理药物)、Aβ诱导组(20μmol/L Aβ处理组)和预处理组(分别加入浓度为5 ml/L、10 ml/L、20 ml/L的丹参川芎嗪注射液孵育24 h后加20μmol/L Aβ),通过CCK-8法检测细胞增殖活性,流式细胞术(FCM)检测细胞凋亡率,Hoechst 33258染色观察PC12细胞核的改变,荧光分光光度计测定LDH、SOD、GSH及caspase-3活性水平,免疫组织化学方法观察细胞色素C(Cyt-C)蛋白释放水平,Western Blot检测Bcl-2的表达水平。结果丹参川芎嗪注射液(5、10、20 ml/L)预处理对Aβ诱导的PC12细胞损伤有较好的保护作用,其保护作用随着药物浓度的增加而增强。它能增加Aβ损伤的PC12细胞增殖活力,减少Aβ诱导的PC12细胞凋亡,降低细胞核凝聚现象,抑制Aβ损伤的PC12细胞LDH释放,增强SOD和GSH活性,促进Cyt-C在细胞内表达,降低caspase-3活性,促进Bcl-2的表达。结论丹参川芎嗪注射液对Aβ诱导的PC12细胞损伤具有与线粒体通路相关的保护作用,其保护作用与它抑制细胞凋亡、抗氧化应激、维持线粒体正常功能、抑制caspase-3的激活、促进抗凋亡因子Bcl-2的表达有关。Objective To observe the possible protective effect and mechanism of Salvia and TMP injection on Aβ-induced injury in PC12 cells.Methods PC12 cells were divided into five groups：control gruop,Aβ-induced Injury in PC12 cells（Treatment with20μmol/L Aβalone）;Pretreatment group（Pretreatment with 5ml/L,10ml/L,20ml/L Salvia and TMP Injection on Aβ-induced Injury in PC12 cells）.Then the cell viability was measured by CKK-8,apoptotic rate was measured by flow cytometry,variation of nucleus was measured by Hoechst33258 dye,and the vitality of LDH,SOD,GSH and caspase-3 were measured by the kits,respectively.Immunohistochemical method was used to observe Cyt-C protein release.And western blot was used to detecting Bcl-2 protein expression.Results Salvia and TMP injection（5,10,20μmol/L）pretreatment can protect the PC12 cells injuried by Aβwhich protective effect was increased with higher drug concentration.Salvia and TMP injection can promote cells proliferation,reduces apoptosis and nuclei condensation,inhibit LDH release,increase SOD,GSH activation,decrease caspase-3 activation,help Cyt-C and Bcl-2 generation.Conclusion Salvia and TMP injection have neuroprotective effect of PC12 cells induced by Aβwhich was related with anti-oxidative stress,maintenance of mitochondrial function,improvement of the anti-apoptotic factor Bcl-2 and inhibition of caspase-3 activation.Salvia and TMP injection may be used to cure neurodegenerative disease such as AD.

关 键 词：丹参川芎嗪注射液 AΒ PC12细胞 凋亡 神经保护 

分 类 号：R741[医药卫生—神经病学与精神病学]