摘要
目的:研究乙醇对氧化应激损伤的作用特点和红景天醇提物的抗氧化作用。方法:采用DPPH自由基生成体系、体外鲁米诺化学发光反应(OH和O_2^-)体系观察红景天醇提物对DPPH自由基、OH和O_2^-化学发光强度的抑制作用以及剂量-效应关系。采用乙醇诱导人正常肝细胞系QZG细胞的氧化应激反应模型,观察红景天醇提物对乙醇诱导细胞活力和氧化应激损伤的保护作用。实验分设红景天醇提物3个不同浓度(50、100、200mg/L)的预防给药组和治疗给药组、阳性对照组(200 mmol/L乙醇干预)和阴性对照组(不加受试物)。预防给药组用红景天醇提物预处理QZG细胞12h后,再加入200mmol/L乙醇处理6h,治疗给药组采用红景天醇提物和乙醇同时处理QZG细胞6h。用MTT试验和生化法测定细胞活力、细胞丙二醛(MDA)、还原型和氧化型谷胱甘肽(GSH、GSSG)和总巯基(T-SH)含量、过氧化氢酶(CAT)、超氧阴离子歧化酶(SOD)活力;免疫印迹法检测细胞抗氧化酶血红素加氧酶-1(HO-1)和核因子相关因子2(NRF-2)蛋白的表达。结果:红景天醇提物对自由基的生成具有显著的抑制作用,呈现较为明显的剂量-效应关系;红景天醇提物可有效保护乙醇所致的肝细胞活力损伤,且治疗组具有明显的剂量-效应关系。与阳性对照组相比,红景天醇提物干预组细胞的MDA含量和GSSG含量下降(P<0.05),GSH和T-SH含量显著升高(P<0.05);CAT和SOD活性也显著升高(P<0.05)。免疫印迹测定结果表明,红景天醇提物可以诱导HO-1蛋白和NRF-2蛋白表达上调(P<0.05)。结论:红景天醇提物在体外自由基模型中具有较强的抗自由基作用,可保护乙醇导致的QZG细胞氧化损伤,其作用机制可能和抗氧化作用相关。
OBJECTIVE：To study the characteristics of alcohol-induced oxidative damage in QZG cells and antioxidant activity of Rhodiola ethanol-extract.METHODS：The DPPH system and chemiluminescence system models for determination of OH and O_2 were established to assess the inhibitive rates of DPPH radicals and chemical luminescence intensity of OH and O_2.The oxidative damage model induced by ethanol in QZG cells was set up to measure the protective activities of Rhodiola ethanol-extract.Ethanol-extract of Rhodiola was divided into 3 different concentrations（50,100,200 mg/L） for the prophylaxis group and treatment groups,positive control group（200 mmol/L ethanol intervention） and negative control group（without test substance）.Prophylaxis QZG cells were pretreated with Rhodiola ethanol-extract for 12 h,then 200 mmol/L ethanol added for 6 h.Treatment group received 200 mmol/L ethanol and Rhodiola ethanol-extract for 6 h.We used MTT test and biochemical method for determining cell vitality, malondialdehyde（MDA） and reduced and oxidized glutathione（GSH,GSSG） and total mercaptoacetic（T-SH） content and catalase（CAT）,superoxide dismutase enzyme（SOD） activity,Western blot to detect protein expression of antioxidant enzymes HO-1 and NRF-2.RESULTS：In the DPPH system and the two chemiluminescence systems,Rhodiola ethanol-extract could significantly inhibit generation of free radicals.In the oxidative damage model induced by ethanol in QZG cells,Rhodiola extract could effectively protect cell injury induced by alcohol,and the treatment groups showed an evident dose-effect relationship.Ethanol-extract of Rhodiola intervention groups reduced the contents of MDA and GSSG compared with positive control group（P0.05）,and increased the content of GSH and T-SH.The treatment groups also demonstrated high CAT and SOD activities.Western blot results showed that Rhodiola ethanol-extract could induce the protein expressions of antioxidant enzymes HO-1 and NRF-2.CONCLUSION：Ethanol-extract of Rhodiola coul
出处
《癌变．畸变．突变》
CAS
CSCD
2011年第2期81-86,共6页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金(30872135)
关键词
红景天醇提物
DPPH
化学发光
乙醇
氧化应激
Rhodiola ethanol-extract
DPPH
chemiluminescence
alcohol
oxidative stress
