千金子提取液对大鼠肺成纤维细胞增殖的影响及细胞毒性作用  

Effects of euphorbiae crude extract on proliferation of lung fibroblasts in rats and its cytotoxicity

作　　者：杨臖 王世岭[1] 付桂英[1] 郭华[1] 吴坤[1] 

机构地区：[1]军事医学科学院附属医院药剂科,北京市100039

出　　处：《中国临床康复》2005年第27期101-103,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:观察原代培养的大鼠肺成纤维细胞给予不同浓度的千金子提取液后,对肺成纤维细胞增殖的影响以及药物的细胞毒性作用。方法:实验于2003-12/2004-03在军事医学科学院附属医院药剂科临床药理室完成。选取纯系Wistar雌性大白鼠10只,千金子为北京首创大地药业有限公司产品,经鉴定为大戟科属植物续随子的种子。经过传代5代以后的肺成纤维细胞大部分呈梭形,可以用于实验,3个实验各自独立。①肺成纤维细胞增殖检测与分组:大鼠肺成纤维细胞在正规生长培养基中传代培养,48h后处于对数生长期,可进行增殖试验。随机分为1个空白对照组和6个实验组。空白对照组加入空白DMEM,实验组分别加入浓度为500,250,125,62.5,31.25,15.625mg/L的千金子提取液。应用四甲基偶氮唑盐比色法观察瑞香狼毒提取液对细胞增殖的影响。②乳酸脱氢酶活性测定与分组:随机分为1个空白对照组和5个实验组。空白对照组加入空白DMEM,实验组分别加入浓度为125,62.5,31.25,15.625,7.813mg/L的千金子提取液。全自动生化分析仪测定乳酸脱氢酶活性值来观察其细胞毒性。③细胞形态学观察与分组:随机分为1个空白对照组和6个实验组。空白对照组加入空白DMEM,实验组分别加入浓度为500,250,125,62.5,31.25,15.625mg/L的千金子提取液,显微镜下观察细胞形态学变化。结果:实验纳入10只大鼠无脱落。①千金子提取液对肺成纤维细胞增殖的影响:浓度为500,250,125,62.5mg/L的千金子提取液的吸光度值明显低于空白对照组(0.108±0.004,0.089±0.002,0.093±0.005,0.187±0.002,0.285±0.011,P<0.01);浓度为31.25,15.625mg/L的千金子提取液的吸光度值仍低于空白对照组(0.231±0.014,0.218±0.023,0.285±0.011,P<0.05)。②千金子提取液对细胞的毒性作用:浓度为15.625,7.813mg/L的千金子提取液其乳酸脱氢酶吸光度值与空白对照组比较,均无显著性差异(28.91±0.88AIM： To observe the influence on the proliferation of lung fibroblasts after the treatment of euphorbiac crude extract of different concentrations, and discuss the cytotoxicity of the medicine. METHODS： The experiment was carried out in the clinical pharmacological room of Department of Pharmacy, Affiliated Hospital, Academy of Military Sciences between December 2003 and March 2004. Ten female pure Wistar white rats were used, euphorbiac was produced by Beijing Shouchuang Dadi Pharmaceutical Company, and it was identifed to be the seed of euphorbiae caper. Most of the lung fibroblasts showed the shape of fusiform after 5 passages, and they could be used in 3 independent experiments. ① Detection of the proliferation of lung fibroblasts and grouping： The lung fibroblasts of rats were subcultured in regular growth medium, and at the logarithm growth period after 48 hours, and could be used in the proliferation test. The lung fibroblasts were randomly divided into one a control group and 6 experimental groups, blank DMEM was added in the blank control group and euphorbiac crude extract of 500, 250, 125, 62.5, 31.25 and 15.625 mg/L were added into the experimental groups respectively. The influence of euphorbiae crude extract on cell proliferation was observed with the methyl-thiazol-tetrazolium colorimetry. ② Detection of the activity of lactate dehydrogenase and grouping： The lung fibroblasts were randomly divided into one a control group and 5 experimental groups, blank DMEM was added in the blank control group and euphorbiac crude extract of 125, 62.5, 31.25, 15.625 and 7.813 mg/L were added into the experimental groups respectively. The activity of lactate dehydrogenase was detected with the automatic biochemistry analyzer to observe its cytotoxicity. ③ Cell morphological observation and grouping： The lung fibroblasts were randomly divided into one a control group and 6 experimental groups, blank DMEM was added in the blank control group and euphorbiae crude extract of 500, 250, 125, 62.5, 31.25 a

关 键 词：细胞分裂 千金子 毒性试验 中药疗法 成纤维细胞 肺 

分 类 号：R563[医药卫生—呼吸系统]