北五味子总木脂素对PC12细胞氧化应激损伤的保护作用及其抑制NF-κB/iNOS/NO信号通路的机制  

Protective effects of Schisandra Chinensis lignans on oxidative stress injury of PC12 cells and its mechanism of inhibition on NF-κB/iNOS/NO signaling pathway

作　　者：姜恩平[1] 于春荣 李贺[3] 于春艳[1,4] 陈建光[1,3] 

机构地区：[1]广东医学院基础医学院肿瘤研究所,广东东莞523808 [2]北京昭衍新药研究中心股份有限公司,北京100176 [3]北华大学药学院药理学教研室,吉林吉林132013 [4]北华大学基础医学院病理学教研室,吉林吉林132013

出　　处：《吉林大学学报（医学版）》2015年第3期532-536,I0006,共6页Journal of Jilin University：Medicine Edition

基　　金：广东医学院博士基金资助课题(B2012063);吉林省教育厅科研基金资助课题(2011130;2012394);吉林省科技厅自然科学基金资助课题(201215103);吉林省吉林市科技局科技计划项目资助课题(201464064)

摘　　要：目的:观察北五味子总木脂素(SCL)对H2O2诱导的PC12细胞氧化应激损伤的保护作用及对NF-κB/iNOS/NO信号通路的影响,并探讨其机制。方法:将处于对数生长期PC12细胞分为对照组、模型组(H2O2200μmol·L-1)、SCL高剂量组(SCL 30mg·L-1+H2O2200μmol·L-1,SCL1组)和SCL低剂量组(SCL 10mg·L-1+H2O2200μmol·L-1,SCL2组)。对照组PC12细胞不做任何处理;模型组PC12细胞用200μmol·L-1 H2O2孵育6h造成细胞氧化应激损伤模型;SCL1和SCL2组分别采用不同剂量SCL预处理PC12细胞2h后再用H2O2(200μmol·L-1)继续孵育6h。MTT法检测各组细胞存活率,酶标仪检测培养基上清中乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)、一氧化氮(NO)水平,流式细胞术检测活性氧(ROS)水平,免疫组织化学和Western blotting法检测PC12细胞中诱导型一氧化氮合酶(iNOS)和核转录因子(NF-κB)P65蛋白的表达情况。结果:与对照组比较,模型组PC12细胞存活率明显降低(P<0.01),细胞上清液中LDH活性升高(P<0.01),SOD活性降低(P<0.01),MDA和NO水平升高(P<0.01),细胞中ROS水平升高(P<0.01),NF-κB阳性细胞数显著增加(P<0.01),iNOS和NF-κB P65蛋白表达水平均升高(P<0.01);与模型组比较,SCL1和SCL2组细胞存活率明显升高(P<0.05或P<0.01),细胞上清中LDH活性降低(P<0.05或P<0.01),SOD活性升高(P<0.01),MDA和NO水平降低(P<0.05或P<0.01),细胞中ROS水平降低(P<0.01),NF-κB阳性细胞数明显减少(P<0.05或P<0.01),iNOS和NF-κB P65蛋白表达水平显著降低(P<0.05)。结论:SCL对氧化应激损伤的PC12细胞具有一定的保护作用,其机制可能与抑制NF-κB/iNOS/NO信号通路、减少氧自由基的产生和减轻脂质过氧化损伤有关联。Objective To investigate the protective effects of Schisandra Chinensis lignans（SCL）on the oxidative stress injury of PC12 cells induced by hydrogen peroxide and influence in NF-κB/iNOS/NO signaling pathway,and to explore the mechanisms.Methods The PC12 cells were divided into control group,model group（H2O2200μmol·L^-1）,high dose of SCL group（SCL 30mg·L^-1＋ H2O2200μmol·L^-1,SCL1group）and low dose of SCL group（SCL 10mg·L^-1＋ H2O2200μmol·L^-1,SCL2 group）.The PC12 cells were untreated（control group）or treated with H2O2（200μmol·L^-1）for 6h（model group）.The PC12 cells in SCL1 and SCL2 group were firstly pretreated with different concentrations of SCL for 2h,and then continuously with SCL and exposed to200μmol·L^-1 H2O2 for 6hat the same time.The survival rate of PC12 cells was evaluated by MTT assay.The activities of lactate dehydrogenase（LDH）,superoxide dismutase（SOD）and the levels of malondialdehyde（MDA）and nitric oxide（NO）were detected by microplate reader.The fluorescence intensity of reactive oxygen species（ROS）was assessed with flow cytometry.Immunohistochemistry and Western blotting methods were used to detect the protein expressions of inducible nitric oxide synthase（iNOS）and nuclear factor kappa B P65（NF-κB P65）.Results Compared with control group,the survival rates of PC12 cells in model group were significantly decreased（P〈0.01）;the LDH activities in the supernatant were increased（P〈0.01）,the SOD activities were decreased（P〈0.01）,the MDA and NO levels were increased（P〈0.01）,the cell ROS level was increased（P〈0.01）,the number of positive cells of NF-κB were increased（P〈0.01）,and the expression levels of iNOS and NF-κB P65 protein were increased（P〈0.01）.Compared with model group,the survival rates of PC12 cells in SCL1 and SCL2groups were increased（P〈0.05 or P〈0.01）,the LDH activities in the supernatant were decreased（P〈0.05 or P〈0.01）,the SOD activities were increased（P〈0.

关 键 词：五味子 氧化应激 核转录因子 一氧化氮合酶 

分 类 号：R285.5[医药卫生—中药学]