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作 者:肖冰[1] 肖丽春 赖卓胜[1] 张亚历[1] 张振书[1] 张万岱[1]
机构地区:[1]中国人民解放军第一军医大学南方医院消化内科,广东省广州市510515 [2]福州市第二人民医院中医科,福建省福州市350007
出 处:《世界华人消化杂志》1999年第11期951-954,共4页World Chinese Journal of Digestology
摘 要:目的 观察自拟治癌方剂镇癌灵诱导胃癌细胞凋亡和抑制胃癌细胞体外生长的效应,为临床应用与新药开发奠定实验基础.方法 镇癌灵是由马钱子、三棱、红根地覃、白花蛇舌草等10余种中药、草药、植物组成的药方. 将镇癌灵按成人每天正常使用剂量500 g( 即10 g/ kg 体重) ,常规煎熬浓缩成500 m L汤液,4 ℃冰箱冷却,0-22 μm 滤膜过滤除菌,以消毒的玻璃离心管分装成每支5 m L,- 20 ℃冰箱储存备用. 根据抗癌药物敏感性实验的方法,计算出体外药物敏感性实验所用药物的一般浓度为5 g/ L,即在含10 % 小牛血清的RPMI1640 完全培养基中加入5 m L/ L的过滤汤剂为中剂量,0-5 m L/ L 和50 m L/ L分别为低剂量和高剂量,常规培养SGC7901 胃癌细胞,3 d 换液一次. 根据直接记数法和MTT 显色法检测细胞生长曲线与细胞增殖实验的同时,每隔36 h 收集细胞进行细胞涂片染色及提取细胞基因组DNA 电泳.结果 生长曲线表明,经高、中、低剂量镇癌灵作用的胃癌细胞实验组的对数生长期分别为3 d ~6-5 d ,3 d ~7 d ,3 d ~7-5 d ,细胞倍增时间各为3 d ,2-5 d ,2-5 d ;而非空白对照组细胞对?AIM To investigate apoptosis and growth inhibition effect of gastric carcinoma cell in vitro induced by the formulae Zhen'ailing made by ourself, and establish the experimental basis for clinical application and development of new drugs. METHODS Zhen'ailing was composed of more than ten types of Chinese herbal medicine and plants including Vomiting nut (马钱子), Burreed tuber (三棱), Sage (红根地覃), oldenlandia (白花蛇舌草), et al . Zhen'ailing was decocted and concentrated in 500*!mL decoction according to the normal dosage of 500*!g ( 10*!g/*!kg body weight) used for adults, cooled at 4℃; bacteria were filtered by 0 22*!μm filtration membrane, then it was put into 5*!mL glass tube, stored at -20℃ refrigerator. The concentration of this formulae was divided into high, moderate and low dose, added by this decoction of 50*!mL/*!L , 5*!mL/*!L , 0 5*!mL/*!L respectively. In the anti tumor drug's sensitive test in vitro SGC7901 stomach carcinoma cell was cultured in this complete medium. The growth curve and proliferation of cells were detected by MTT test and direct counting method, Simultaneously cells were collected, smeared and stained, the genomic DNA was extracted for gel electrophoresis once every 36 hours. RESULTS The growth curve indicated that the logarithmic growth phase of the experimental group of stomach carcinoma cells were 3*!d - 6 5*!d , 3*!d - 7*!d , 3*!d - 7 5*!d , respectively by high, moderate and low dose of Zhen'ailing, but the control group was 2*!d - 8*!d and the doubling time was 2*!d . The results of MTT test showed the cell proliferation rate or proliferation index and survival rate of experimental group by high, moderate and low dose of Zhen'ailing were lower than control group. There was significant difference between the two groups ( P <0 01). The staining of brush cells of experimental group showed nuclear shrinkage, margination, nuclear fragmentation and formation of apoptotic bodie
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