银杏二萜内酯葡胺注射液通过下调calpain信号通路抑制脑缺血神经细胞凋亡  

Diterpene ginkgolides meglumine injection attenuates oxygen-glucose deprivation-induced apoptosis of nerve cells via inhibition of calpain signaling pathway

作　　者：刘秋[1] 许治良[1] 金治全[1] 周军[1] 毕宇安[1] 王振中[1] 萧伟[1] 

机构地区：[1]江苏康缘药业股份有限公司中药制药过程新技术国家重点实验室,连云港222001

出　　处：《中国药科大学学报》2015年第6期707-711,共5页Journal of China Pharmaceutical University

基　　金：国家"重大新药创制"科技部重大专项资助项目(No.2013ZX09402203);国家重大科技成果转化项目基金资助~~

摘　　要：研究银杏二萜内酯葡胺注射液(diterpene ginkgolides meglumine injection DGMI)对缺氧缺糖/复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤的人神经母细胞瘤细胞SH-SY5Y凋亡的抑制作用及其机制。采用试剂盒、Fluo-3 AM法、Western blot检测SH-SY5Y细胞氧糖剥离损伤4h后,与药物一起复氧1h细胞乳酸脱氢酶(LDH)的漏出量、caspase-3/7酶活力、细胞质中核小体含量、胞浆游离钙离子浓度以及calpain、cleaved caspase-12蛋白量的变化。结果显示DGMI能极显著降低OGD/R损伤的SH-SY5Y细胞LDH的漏出量,抑制caspase-3/7酶活性,减少细胞核核小体的释放量,下调细胞内游离钙离子浓度、calpain和cleaved caspase-12蛋白量,抑制calpain凋亡信号通路保护神经细胞。DGMI对OGD/R诱导的SH-SY5Y细胞凋亡具有显著的抑制作用,其作用机制可能与抑制细胞内Ca^(2+)/calpain/caspase-12/capase-3信号通路有关。To investigate the anti-apoptotic effect of diterpene ginkgolides meglmnine injection （DGMI） on SH- SY5Y ceils induced by oxygen-glucose deprivation/reoxygenation （ OGD/R）, and to explore its mechanisms. After 4 h of OGD, the SH-SY5Y cells were treated with 25 mg/L DGMI for 1 h. The release of lactic dehydrogenase （LDH） was measured by cytotoxicity detection kitCus. Cell apoptosis was detected by easpase-3/7 assays. Cell death was detected by ELISA. The concentration of [ Ca2 ＋ ] in cytoplasm was measured by Fluo-3 AM and the levels of ealpain and cleaved capaease-12 were evaluated by western blot. As we expected, DGMI significantly decreased the release of LDH, the concentration of [ Ca2 ＋1 ], the protein levels of ealpain and cleaved caspase-12. Furthermore, DGMI injection also attenuated the activities of caspase-3/7 and the contents of cytoplasmic histoneassociated- DNA-fragments. These data demonstrated that the DGMI injection showed good anti-apoptotic effect in SH-SY5Y cells induced by OGD/R.The mechanisms may be associated with the inhibition of Ca2＋/calpain/ caspase-12/caspase-3 signaling pathway.

关 键 词：银杏二萜内酯葡胺注射液 缺氧缺糖损伤 细胞凋亡 钙离子 CALPAIN caspase-12 

分 类 号：R285[医药卫生—中药学]