基于Nrf2信号通路的三七总皂苷对Aβ_(25-35)诱导PC12细胞凋亡的保护作用机制研究  

Protective effect of Panax Notoginseng Saponins of Nrf2 signaling pathway on apoptosis of PC12 cells induced by Aβ_(25-35)

作　　者：赵静宇[1,2] 汪梦霞[1,2] 赵自明 孟祥宝[3] 孙桂波[3] 孙晓波[3] 

机构地区：[1]广州中医药大学附属广东省第二中医院,广东广州510405 [2]广东省中医药工程技术研究院广东省中医药研究开发重点实验室,广东广州510095 [3]中国医学科学院北京协和医学院药用植物研究所中药(天然产物)创新药物研发北京市重点实验室,北京100193

出　　处：《中国药理学通报》2016年第3期343-349,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81503290);中医药行业科研专项(No 201507004);国家重大创新药创制专项(No2012ZX09501001-004)

摘　　要：目的探讨三七总皂苷对Aβ_(25-35)诱导的PC12细胞凋亡的保护作用及分子机制。方法利用Aβ_(25-35)诱导PC12细胞损伤模型,MTT法检测PC12细胞活力,试剂盒测定LDH漏出量、ROS水平、MDA含量、Caspase-3活力以及抗氧化酶SOD、CAT和GHS-Px活力,TUNEL染色检测细胞凋亡,JC-1染色观察线粒体膜电位,Western blot检测相关蛋白HO-1、Lamin-B、细胞核内Nrf2及总Nrf2的蛋白表达。结果三七总皂苷能够明显抑制Aβ_(25-35)诱导的PC12细胞活力下降(P<0.01)和LDH漏出(P<0.01),减少ROS和MDA的产生(P<0.01),增加SOD、CAT和GSH-Px活力(P<0.01),稳定线粒体膜电位,抑制caspase-3的活化(P<0.01)。而且,三七总皂苷还能够上调PC12细胞细胞核Nrf2、总Nrf2和细胞质中HO-1的蛋白表达。HO-1抑制剂Sn PP能够抑制三七总皂苷的神经保护作用。结论三七总皂苷可能通过上调Nrf2/HO-1信号通路,从而抑制Aβ25-35诱导PC12细胞氧化应激和凋亡。Aim To investigate the protective effect of Panax Notoginseng Saponins(PNS) on Aβ_(25-35)-induced apoptosis in PC12 cells and molecular mechanism.Methods The cell viability of PC12 cells was detected by MTT assay.The levels of LDH leakage,ROS,MDA,Caspase-3 activity and antioxidant enzyme activities of SOD,CAT,and GSH-Px activity were determined by respective kits.The apoptosis of cells was decteced by Western blot.Results PNS could significantly inhibit the decrease of cell viability and LDH leakage,reduce the production of MDA and ROS(P<0.01),increase the activity of SOD,CAT and GSH-Px(P<0.01),and the mitochondrial membrane potential,inhibit the activation of caspase-3(P<0.01) in PC12 cells which were induced by Aβ_(25-35).PNS could incrase nuclear Nrf2 and up-regulate HO-1.The neuroprotective of PNS could be inhibited by HO-1 inhibitor Zn PP.Conclusion PNS may inhibit Aβ_(25-35)-induced oxidative stress and apoptosis in PC12 cells by activating Nrf2/HO-1 signaling pathway.

关 键 词：阿尔茨海默病 三七总皂苷 氧化应激 凋亡 NRF2 HO 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学]