红景天苷通过Nrf2-HO-1保护Aβ25-35诱导的原代皮层神经元损伤  

Attenuation of Aβ-induced Neurotoxicity by Salidroside Through Activation of Nrf2/HO-1 Pathway

作　　者：张霜梅 祝维峰 王安荣 李衍滨[5] 张兆旭 ZHANG Shuangmei;ZHU Weifeng;WANG Anrong;LI Yanbin;ZHANG Zhaxu(The Cancer Hospital of the University of Chinese Academy of Sciences,Zhejiang Cancer Hospital,Hangzhou 310022,Zhejiang,China;Institute of Basic Medicine and Cancer(IBMC)Chinese Academy of Sciences,Hangzhou 310022,Zhejiang,China;Chinese Medicine Hospital Affiliated to Guangzhou Medical University,Guangzhou 510006,Guangdong,China;Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China;Shandong Province Qianfoshan Hospital,Shandong First Medical University,Jinan 250014,Shandong,China;People's Hospital of Peking University,Beijing 100044,China)

机构地区：[1]中国科学院大学附属肿瘤医院,浙江省肿瘤医院,浙江杭州310022 [2]中国科学院基础医学与肿瘤研究所,浙江杭州310022 [3]广州医科大学附属中医医院,广东广州510006 [4]广州中医药大学,广东广州510006 [5]山东第一医科大学附属山东省千佛山医院,山东济南250355 [6]北京大学人民医院,北京100044

出　　处：《中华中医药学刊》2021年第1期101-106,I0031,I0032,I0033,I0034,I0035,I0036,共12页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金(81303013);祝维峰广东省中医药管理局名中医工作室建设项目(粤中医办函[2018]5号)。

摘　　要：目的探讨红景天苷对Aβ25-35诱导的皮层神经元损伤的保护作用。方法:随机将培养成熟的原代皮层神经分为5组:正常对照组、模型组、红景天苷5μmol·L^(-1)组、红景天苷10μmol·L^(-1)组、激动剂组。后3组分别使用5μmol·L^(-1)红景天苷、10μmol·L^(-1)红景天苷、10μmol·L^(-1)TBHQ培养24 h,余组正常换液。然后除正常对照组,将余组更换为终浓度为20μmol·L^(-1)Aβ25-35并维持各药物组终浓度的培养液,24 h后使用CCK8试剂盒测细胞活性、LDH试剂盒测细胞LDH释放量、免疫组化和Western Blot测Nrf2、HO-1、Caspase-3、Bax、Bcl-2等蛋白的表达水平。结果红景天苷5μmol·L^(-1)组、红景天苷10μmol·L^(-1)组、TBHQ组都能够提高Aβ25-35损伤后细胞的活性、减少LDH的释放改善细胞损伤,红景天苷10μmol·L^(-1)组、TBHQ组都能够增加Nrf2、HO-1的表达水平,下调Caspase-3蛋白表达水平、上调Bcl-2/Bax的蛋白表达水平。锌原卟啉ZnPP可逆转红景天苷及TBHQ对Aβ25-35诱导的皮层神经元的保护作用。结论红景天苷对Aβ25-35诱导的皮层神经元细胞的保护作用,可能主要通过改善凋亡相关蛋白、上调Nrf2/HO-1信号通路表达实现。Objective To investigate the protective effect of salidroside on cortical neuron damage induced by Aβ25-35.Methods The cultured and mature primary cortical nerves were randomly divided into 5 groups:normal control group,model group,salidroside 5μmol·L^(-1)group,salidroside 10μmol·L^(-1)group and agonist group.The last three groups were cultured with 5μmol·L^(-1)salidroside,10μmol·L^(-1)salidroside and 10μmol·L^(-1)TBHQ for 24 h,and the remaining groups were replaced the cultural liquid normally.And then except the normal control group,the remaining groups were replaced with a culture medium with a final concentration of 20μmol·L^(-1)Aβ25-35 and the final concentration of each drug group was maintained.After 24 hours,the CCK8 kit was used to measure cell viability,and the LDH kit was used to measure cell LDH release and immunity histochemistry and Western blot measured the expression levels of Nrf2,HO-1,Caspase-3,Bax,Bcl-2 and other proteins.Results Salidroside 5μmol·L^(-1)group,salidroside 10μmol·L^(-1)group and TBHQ group can increase the activity of cells after Aβ25-35 injury,reduce the release of LDH and improve cell damage.Salidroside 10μmol·L^(-1)group and TBHQ group can increase the expressions of Nrf2 and HO-1,down-regulate the protein expressions of Caspase-3,and up-regulate the protein expressions of Bcl-2/Bax.Zinc protoporphyrin(ZnPP)can reverse the protective effects of salidroside and TBHQ on cortical neurons induced by Aβ25-35.Conclusion The protective effect of salidroside on cortical neurons induced by Aβ25-35 may be mainly achieved by improving the expressions of apoptosis-related proteins and up-regulating the expression of Nrf2/HO-1 signaling pathway.

关 键 词：红景天苷 阿尔茨海默病 氧化应激 NRF2 HO-1 凋亡蛋白 

分 类 号：R285.5[医药卫生—中药学]