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 bodies, and the characteristic ladder band on gel electrophoresis. NO changes were observed in control group. CONCLUSION Zhen'ailing has multiple functions including promoting blood circulation, promoting salivation, removing heat, supplementing qi. The results of this experiment showed that the proliferation of stomach carcinoma cells cultured in vitro was inhibited by this formulae Zhen'ailing and the mechanism was probably associated with the induction of apoptosis of stomach carcinoma cells.
出处
《世界华人消化杂志》
CAS
1999年第11期951-954,共4页
World Chinese Journal of Digestology
关键词
镇癌灵
药理学
细胞凋亡
胃肿瘤
Zhen'ailing/pharmacology
apoptosis
stomach neoplasm
