金银花对大鼠体内和体外RBL细胞的抗氧化保护作用及其机制  

Antioxidant effect of honeysuckle on hepatic RBL cells of rats in vivo and in vitro and mechanism

作　　者：宫璀璀[1] 郑乃刚[2,3] 吴景兰[3] 赖泽仁[1] 何培霞[1] 

机构地区：[1]中国人民解放军153医院检验中心,河南郑州450042 [2]郑州大学基础医学院肿瘤基础学教研室,河南郑州450001 [3]郑州大学基础医学院分子细胞生物学研究中心,河南郑州450042

出　　处：《吉林大学学报（医学版）》2009年第6期1074-1078,共5页Journal of Jilin University：Medicine Edition

基　　金：河南省科技厅基础与前沿技术研究计划项目资助课题(072300410440)

摘　　要：目的:从细胞和分子水平探讨金银花(HS)对机体内外的抗氧化作用及其机制,为金银花的临床应用提供理论依据。方法:体外培养的人肝RBL细胞与0.25g·L-1的金银花共培养前或后加H2O2(氧化剂),应用MTT和TUNEL实验检测细胞生存率和凋亡率,免疫组织化学/免疫印迹检测NF-κB、HSP-70、Bcl-2、Bax及Caspase-3表达水平,检测细胞内抗氧化酶体系的水平。建立金银花大、小剂量灌胃的大鼠动物模型,检测血浆抗氧化酶体系的总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、谷胱苷肽过氧化物酶(GSH-Px)、谷胱苷肽(GSH)、丙二醛(MDA)的水平。结果:金银花对RBL细胞的抗氧化和抗凋亡作用中,前保护(先加金银花,后加H2O2)高于后保护(先加H2O2,后加金银花)的效应;下调NF-κB和HSP-70的表达和Bcl-2/Bax比值,且前保护组凋亡率低于后保护组;同时也上调细胞内抗氧化酶体系的水平。金银花灌胃1～2h后,大鼠血浆中T-AOC、GSH-Px、GSH、SOD含量较灌胃前显著增高(P<0.05),而MDA含量明显减低(P<0.05)。结论:金银花可通过上调体内抗氧化酶体系并下调NF-kB信号传导途径呈现抗氧化、抗凋亡的保护作用。Objective To explore the antioxidant effect of honeysuckle on hepatic RBL cells of rats in vivo and in vitro and its underlying mechanism,and provide theoretical basis for the application of honeysuckle in clinic.Methods The blood levels of T-AOC,SOD,GSH-Px,GSH or MDA in an antioxidant enzyme system were detected 1-2 h after administration in the established rat models,which were administrated with larger and smaller doses of honeysuckle respectively.The cultured human hepatic RBL cells were exposed to H2O2 as an oxidant and treated with 0.25g·L-1 neysuckle before or after exposure to H2O2.The cell survival and apoptosis were detected by MTT and TUNEL assays.The expression levels of NF-kB,Bcl-2,Bax,HSP-70 and Caspase-3 were determined by immunohistochmistry and Western blotting and the cell anti-oxidant enzyme system was detected by corresponding kits.Results In the antioxidant and anti-apoptotic effects of honeysuckle,the pre-protective effect was markedly higher than the post-protective.The expressions of NF-κB and HSP-70,the expression ratio of Bcl-2/Bax were down-regulated,and the apoptotic rate in pre-protective group was lower than that in post-protective group. Besides,the detected cell level of anti-oxidant enzyme system was up-regulated significantly.In the plasma of rat model the levels of T-AOC,GSH-Px,GSH and SOD were promoted（P〈0.05）,while that of MDA was decreased markedly（P〈0.05）.Conclusion Honeysuckle displays antioxidant capacity through up-regulating the antioxidant enzyme system and down-regulating NF-κB signal transduction pathway.

关 键 词：金银花 过氧化氢 抗氧化酶体系 抗凋亡信号传导 人肝RBL细胞系 

分 类 号：R285.5[医药卫生—中药学] R329.28[医药卫生—中医学]