四氢小檗碱对过氧化氢致前成骨细胞MC3T3-E1氧化损伤的防治作用及机制探讨  

Protective Effect of Tetrahydroberberine on H_2O_2-induced Damage in MC3T3-E1 in vitro

作　　者：朱建红[1] 薛照芸 黄光业 陈健文[2] 石勇辉 伍俊妍[1] ZHU Jianhong;XUE Zhaoyun;HUANG Guangye;CHEN Jianwen;SHI Yonghui;WU Junyan(Department of Pharmacy,Sun-Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou,Guangdong 510120,China;School of Pharmaceutical Sciences,Sun-Yat-sen University,Guangzhou,Guangdong 510006,China)

机构地区：[1]中山大学孙逸仙纪念医院药学部,广东广州510120 [2]中山大学药学院药理与毒理学实验室,广东广州510006

出　　处：《今日药学》2018年第10期649-654,共6页Pharmacy Today

基　　金：广东省科技计划项目(2014A020212134)

摘　　要：目的观察四氢小檗碱对过氧化氢致前成骨细胞MC3T3-E1氧化损伤的影响,并探讨相关机制。方法建立体外MC3T3-E1细胞氧化应激模型,将细胞随机分为H_2O_2+1μmol·L^(-1)四氢小檗碱组; H_2O_2+0.1μmol·L^(-1)四氢小檗碱组; H_2O_2+0.01μmol·L^(-1)四氢小檗碱组; H_2O_2+N-乙酰半胱氨酸(NAC) 1 mmol·L^(-1)组; H_2O_2模型对照组;空白对照组。MTT法检测细胞活力;荧光显微镜观察活性氧(ROS)含量;比色法检测丙二醛(MDA)含量;流式细胞术检测细胞凋亡率; Real time-PCR和Western-blot检测凋亡相关蛋白Bcl-2、Bax表达情况。结果与空白对照组比较,单纯H_2O_2处理组细胞活性明显降低,凋亡率及ROS、MDA含量明显增加;同时Bcl-2 mRNA和蛋白表达明显下调,Bax mRNA和蛋白表达明显上调(P<0.05)。与H_2O_2模型对照组比较,四氢小檗碱能够明显增强细胞活性,降低细胞凋亡率及ROS、MDA含量,上调Bcl-2 mRNA和蛋白表达,下调Bax mRNA和蛋白表达,并且呈剂量依赖性(P<0.05)。结论四氢小檗碱对H_2O_2介导MC3T3-E1细胞的氧化损伤具有保护作用,其机制可能与降低细胞内MDA和ROS的水平相关。OBJECTIVE To study the protective effect of tetrahydroberberine on H2O2-induced damage in MC3T3-E1,and its action mechanism. METHODS The oxidative stress cell model was established by using hydrogen peroxide（ H2O2）,and divided into 6 groups： tetrahydroberberine 1 μmol·L^-1 group,tetrahydroberberine 0.1 μmol·L^-1 group,tetrahydroberberine 0.01 μmol·L^-1 group,NAC1 mmol·L^-1 group,H2O2 model group,and control group. The cell viabilities were detected by MTT assay,and cell apoptosis rates were detected by flow cytometry.The content of malondialdehyde（ MDA） and reactive oxygen species（ ROS） were measured by fluorescence microscopy and colorimetric technique,respectively. The mRNA and protein expression levels of Bcl-2 and Bax were detected by Real time-PCR and Western Blot,respectively. RESULTS Compared with those in the H2O2 model group,tetrahydroberberine 0. 01-1 μmol·L^-1 significantly increased the cell viabilities,and decreased the cell apoptosis rates and the levels of ROS and MDA（ P〈0.05）.In addition,the expression levels of Bcl-2 mRNA and protein were obviously up-regulated,and the expression levels of Bax mRNA and protein were significantly down-regulated,with a dose-dependent manner（ P〈0. 05）. CONCLUSION Tetrahydroberberine can prevent MC3 T3-E1 from oxidative damage in vitro.

关 键 词：四氢小檗碱 MC3T3-E1细胞 氧化应激 

分 类 号：R963[医药卫生—微生物与生化药学]