小檗碱与5'-N-乙基甲酰胺基腺苷单独及联合给药对心肌细胞缺氧/复氧损伤的影响  

Effects of alone and co-administration of berberine and5′-N-ethylformamidoadenosine on cardiomyocyte hypoxia/reoxygen injury

作　　者：贡美娜 高亚云 张束滢 逄晓倩 田炜 徐菁蔓 GONG Mei-na;GAO Ya-yun;ZHANG Shu-ying;PANG Xiao-qian;TIAN Wei;XU Jing-man(School of Public Health,North China University of Science and Technology;Hebei Key Laboratory of Organ Fibrosis;Laboratory Animal Center of North China University of Science and Technology,Tangshan Hebei 063000,China)

机构地区：[1]华北理工大学公共卫生学院 [2]河北省器官纤维化重点实验室 [3]华北理工大学实验动物中心,河北唐山063000

出　　处：《中国药理学通报》2024年第12期2311-2318,共8页Chinese Pharmacological Bulletin

基　　金：中央引导地方科技发展资金项目(No 226Z7711G);河北省引进留学人员资助项目(No C20220354);华北理工大学公共卫生学院青年人才托举计划(No QNRC202313)。

摘　　要：目的 探讨小檗碱(berberine,BBR)与5'-N-乙基甲酰胺基腺苷(5'-n-ethylformamidoadenosine,NECA)单独及联合给药对缺氧/复氧(H/R)诱发的心肌H9c2和HL-1细胞损伤的影响。方法 心肌H9c2和HL-1细胞分为Control组、BBR组、NECA组、联合给药组、H/R组、BBR+H/R组、NECA+H/R组、联合给药+H/R组,CCK-8检测细胞活力;TMRE检测MMP;DCFH-DA检测ROS的含量;Mito SOX Red探针检测线粒体超氧化物;Western blot检测COXⅣ、Tom20、Tim23蛋白表达;qRT-PCR检测COXⅣ、Tom20基因表达。结果 在H9c2细胞中,与Control组相比,H/R组细胞活力、TMRE荧光强度明显降低,COXⅣ、Tom20和Tim23蛋白表达、COXⅣ和Tom20基因表达、ROS和线粒体超氧化物的含量均明显升高。与H/R组相比,BBR和NECA单独给药后细胞活力均增强;单独给药及联合给药后,ROS和线粒体超氧化物的含量明显降低。在HL-1细胞中,与Control组相比,H/R组细胞活力明显降低;ROS和线粒体超氧化物的含量均明显升高。与H/R组相比,BBR和NECA单独给药及联合给药后,细胞活力均增强;ROS和线粒体超氧化物的含量均明显降低。结论 BBR和NECA单独给药及联合给药是通过减少线粒体超氧化物和细胞ROS的生成,进而缓解线粒体损伤,减轻氧化应激损伤,最终降低H/R诱发的心肌细胞损伤。Aim To investigate the effects of berberine(BBR)combined with 5′-n-ethylformamidoadenosine(NECA)on myocardial H9c2 and HL-1 cell damage induced by hypoxia/reoxygenation(H/R).Methods H9c2 and HL-1 cells were divided into the Control group,BBR group,NECA group,combined administration group,H/R group,BBR+H/R group,NECA+H/R group,and combined administration+H/R group.CCK-8 was used to detect cell viability in each group.The TMRE kit was used to detect MMP.DCFH-DA was used to detect ROS content.The Mito SOX Red fluorescent probe was used to detect mitochondrial superoxide.The expressions of COXⅣ,Tom20,and Tim23 were detected by Western blot.The expression of COXⅣand Tom20 genes was detected by qRT-PCR.Results In H9c2 cells,the cell viability and TMRE fluorescence intensity in the H/R group were significantly decreased compared with the Control group.The protein expressions of COXⅣ,Tom20,and Tim23,gene expressions of COXⅣand Tom20,ROS,and mitochondrial superoxide contents were significantly increased.Compared with the H/R group,the cell viability of BBR and NECA were enhanced after administration alone.The contents of ROS and mitochondrial superoxide were significantly decreased.In HL-1 cells,cell viability in the H/R group was significantly decreased compared with the Control group.The contents of ROS and mitochondrial superoxide were significantly increased.Compared with the H/R group,BBR and NECA alone and combined administration enhanced cell viability.The contents of ROS and mitochondrial superoxide were significantly decreased.Conclusion The administration of BBR and NECA alone or in combination can reduce the production of mitochondrial superoxide and cell ROS,thereby alleviating mitochondrial damage,alleviating oxidative stress damage,and ultimately reducing H/R-induced myocardial cell damage.

关 键 词：心肌缺氧/复氧损伤 小檗碱 NECA 线粒体超氧化物 ROS 氧化应激 

分 类 号：R-332[医药卫生] R284.1R329.24R329.411R342.5R542.2