白藜芦醇抑制血管紧张素Ⅱ诱导的大鼠心脏成纤维细胞增殖的实验研究  

Inhibitory Effect of Resveratrol on Cardiac Fibroblast Proliferation Induced by Angiotensin Ⅱ

作　　者：王世君[1] 王兴祥[1] 范芳华[2] 严杰[2] 刘云英[2] 范钰[3] 陈君柱[1] 

机构地区：[1]浙江大学医学院附属第一医院心内科,杭州310003 [2]浙江大学病原微生物实验室 [3]浙江大学肿瘤研究所

出　　处：《中国中西医结合杂志》2008年第4期334-338,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省中医药重点项目基金资助(No.2007ZA015)

摘　　要：目的观察白藜芦醇对血管紧张素Ⅱ（AngⅡ）诱导的大鼠心脏成纤维细胞（cFs）增殖的影响及其机制。方法差速贴壁法培养新生大鼠cFs，在体外建立AngⅡ诱导的cFs增殖模型。采用MTT法检测细胞增殖，分别观察一氧化氮合酶抑制剂（L-NAME）、鸟苷酸环化酶抑制剂（ODQ）及白藜芦醇对AngⅡ诱导的cFs增殖的影响；采用心钠素（ANP）及脑钠素（BNP）mRNA表达检测细胞肥厚反应；放免法及ELISA法测细胞培养液中ANP及BNP水平；RT-PCR方法检测ANP、BNPmRNA表达；硝酸还原酶法测细胞培养液中一氧化氮（NO）水平；化学比色法测细胞上清液中一氧化氮合酶（NOS）水平；放免法测定细胞内环磷酸鸟苷（cGMP）水平。结果白藜芦醇25-100μmol／L呈时间及剂量依赖性抑制AngⅡ诱导的cFs增殖，但这种作用可被L-NAME及ODQ部分阻断。白藜芦醇作用细胞后NO、cGMP水平升高，ANP、BNP水平降低，ANP、BNPmRNA的表达下降。结论白藜芦醇在一定浓度范围对AngⅡ诱导的cFs增殖及细胞肥厚有抑制作用。上调NO-cGMP信号通路可能是其发挥作用的途径之一。Objective To observe the effect and mechanism of resveratrol on cardiac fibroblast （cFs） proliferation induced by angiotensin Ⅱ （Ang Ⅱ ）. Methods The in vitro cFs proliferation model was established by stimulating cultured cFs of new born rats with Ang Ⅱ by differential attachment method. Cell proliferation was measured by MTY assay, and the effect of resveratrol, L-NAME and ODQ on cell proliferation were observed respectively. Besides, the hypertrophic response of cFs was estimated by measuring expressions of atrial natriuretic peptide （ANP） and brain natriuretic peptide （BNP） mRNA, with the levels of ANP and BNP in culture medium determined by radioimmunoassay and ELISA respectively; and their mRNA expressions determined by reverse transcription polymerase chain reaction （RT-PCR）. Level of nitric oxide （NO） in the culture medium was measured by Griess reagent; nitric oxide synthase （NOS） level by chemical colorimetric method; and cGMP by radioimmunoassay. Results Resveratrol at the dose of 25-100 μmol/L inhibited cFs proliferation in a time and dose dependent manner, which could be partially blocked by pretreatment with L-NAME or ODQ. NO and cGMP levels increased, ANP, BNP levels and their mRNA expression lowered after resveratrol treatment. Conclusion Resveratrol in a definite concentration range could inhibit cFs proliferation and hypertrophic response induced by Ang Ⅱ, up-regulating the signal pathway of NO and cGMP might be one of the acting paths of the inhibitory effects.

关 键 词：白藜芦醇 心脏成纤维细胞 心钠素 脑钠素 一氧化氮 环磷酸鸟苷 

分 类 号：R285.5[医药卫生—中药学]