白藜芦醇对人胚肺成纤维细胞株MRC-5生长的抑制作用  

Inhibitory effects of resveratrol on human pulmonary fibroblast line MRC-5 growth

作　　者：刘理静 钱红 张平[2] 王在岩[2] 

机构地区：[1]湖南医药学院临床医学系,湖南怀化418000 [2]南华大学附属第一医院呼吸内科,湖南衡阳421001

出　　处：《中国药理学通报》2015年第5期673-679,共7页Chinese Pharmacological Bulletin

基　　金：湖南省教育厅重点资助项目(No 10A107);湖南省教育厅优秀青年资助项目(No 14B142)

摘　　要：目的探讨白藜芦醇(Res)对人胚肺成纤维细胞生长的抑制作用及相关机制。方法以人胚肺成纤维细胞MRC-5为研究对象,分别予20μL二甲基亚砜(DMSO)及0、12.5、25、50、100、200μmol·L-1Res处理细胞24、48、72 h,通过MTT法分析细胞增殖抑制率。另外,以20μL DMSO(溶媒组)及50、100μmol·L-1Res孵育细胞48 h,采用流式细胞术检测细胞周期和凋亡率,行原位缺口末端标记法(TUNEL)测定细胞凋亡指数(AI),应用荧光实时定量PCR和Western blot分别检测细胞周期蛋白D1(cell cycle protein D1,Cyclin D1)、细胞周期蛋白依赖激酶4(cyclin-dependent kinase 4,CDK4)mRNA与蛋白表达,Western blot检测Bcl-2、Bax蛋白表达。结果随着Res浓度的升高和处理时间的延长,细胞增殖抑制率逐渐增加(P<0.01)。在共同培养48h后,50、100μmol·L-1Res处理组S、G2/M期DNA比例及Cyclin D1、CDK4 mRNA与蛋白表达水平、Bcl-2蛋白表达水平降低,而G0/G1期DNA比例、AI、凋亡率及Bax蛋白表达水平增加,与溶媒组比较,差异均有显著性(P<0.01),并且100μmol·L-1Res效果强于50μmol·L-1(P<0.01)。结论Res能抑制MRC-5细胞增殖,其机制可能与阻碍细胞周期进展及促进细胞凋亡有关。Aim To explore the inhibitory effects of resveratrol （Res ） on human pulmonary fibroblast growth,and its related mechanisms.Methods Hu-man pulmonary fibroblasts MRC-5 were cultured in vitro as research object.These cells were inoculated＆nbsp;with 20 μL dimethyl sulfoxide （DMSO）as well as 0, 12.5,25 50,100 and 200 μmol·L-1 Res for 24,48 and 72 h,respectively.Inhibitory rate of cellular pro-liferation was analyzed by MTT.In addition,these cells were treated with 20 μL DMSO （medium group）＆nbsp;as well as 50 and 100 μmol·L-1 Res for 48 h,re-spectively.Subsequently,cell cycle and apoptotic rate were measured using flow cytometry.Apoptosis index （AI ） was detected through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL）. The mRNA and protein expression of cell cycle protein D1 （Cyclin D1 ） and cyclin-dependent kinase 4 （CDK4）was detected through fluorescence real-time quantitative PCR and Western blot, respectively. Western blot was used to measure the protein expres-sion of Bcl-2 and Bax.Results With the increase of Res concentrations and prolongation of treated time, inhibitory rate of cellular proliferation was gradually el-evated （P〈0.01）.After 48 h of co-culture,DNA ra-tio of S and G2/M periods,mRNA and protein levels of Cyclin D1 and CDK4,and Bcl-2 protein levels were significantly decreased while DNA ratio of G0/G1 peri-od,AI,apoptotic rate and Bax protein levels were sig-nificantly increased in 50 and 100 μmol · L-1 Res-treated groups as compared to medium group （P 〈0.01 ）.Moreover,the effects 100 μmol · L-1 Res were better than those of 50 μmol · L-1 Res （P 〈0.01）.Conclusion Res can suppress the prolifera-tion of MRC-5 cells,which may be associated with blockade of cell cycle progression and induction of cell apoptosis.

关 键 词：白藜芦醇 人胚肺成纤维细胞 增殖 细胞周期 凋亡 细胞周期蛋白D1 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]