中药蛇六谷提取物对人胃癌SGC-7901细胞增殖影响的实验研究(1)
摘 要：目的：观察蛇六谷提取物对人胃癌SGC-7901细胞增殖的影响，探讨其可能的作用机理。方法：采用系统溶剂法初步提取分离蛇六谷不同提取物，运用MTT法和生长曲线法观察不同浓度的各提取物对人胃癌SGC-7901细胞的增殖的影响，流式细胞仪检测蛇六谷石油醚萃取物对该细胞周期分布及凋亡的影响。结果：蛇六谷醇提取物、石油醚萃取物和乙酸乙酯萃取物均具有一定的抑制人胃癌SGC-7901细胞增殖的作用，以石油醚萃取物的抑制作用最为明显，呈现较好的剂量-效应曲线。生长曲线法观察发现蛇六谷石油醚萃取物、乙酸乙酯萃取物对人胃癌SGC-7901细胞增殖抑制作用有一定的时间-效应曲线；流式细胞仪检测发现蛇六谷石油醚萃取物可阻滞SGC-7901细胞周期于G0/G1期，分析表明其作用机理可能与诱导肿瘤细胞凋亡有关。结论：蛇六谷石油醚萃取物可能是该药抗肿瘤的有效部位之一。
关键词：蛇六谷；提取物；人胃癌SGC-7901细胞；细胞增殖；细胞周期；凋亡
, http://www.100md.com
中图分类号：R285.5文献标识码：A
文章编号：1673-7717(2008)11-2313-04
Empirical Study on Effect of Different Extracts of Rhizoma Amorphophalli on the Proliferation of Cell SGC-7901
CHEN Peifeng1,WU Qiaofeng2,DING Zhishan2, YU Ying1, PAN Lei1
(1.The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,Zhejiang,China;
, 百拇医药
2.ZheJiang Chinese Medical University,Hangzhou 310053,Zhejiang,China)
Abstract:Objective:To observe the effect of different extracts of Rhizoma Amorphophalli on the proliferation of cell SGC-7901,and explore the possibility mechanism of them.Methods:Different extracts of Rhizoma Amorphophalli were extracted by the technique of solvent-refining; effect of different dose groups of extracts on cell SGC-7901 were tested by MTT and growth curve method; effects of the extracts of petroleum ether on cell cycle and apoptosis were inspected by flow cytometry.Results:The extracts of ethanol, petroleum ether and ethyl acetate of Rhizoma Amorphophalli have inhibitory effect on the proliferation of cell SGC-7901,and the extract of petroleum ether has much more evident effect, showed a dose-effect curve. Growth curve method showed the extract of petroleum ether and ethyl acetate of Rhizoma Amorphophalli had a time-effect effect on the proliferation of cell SGC-7901. Flow cytometry detection showed the extract of petroleum ether of Rhizoma Amorphophalli can block the cell cycle of SGC-7901 at G0/G1 period,and the mechanism possibly related to inducing apoptosis of tumor cell.Conclusion:The extract of petroleum ether of Rhizoma Amorphophalli is possibly one of antineoplastic parts of the plant.
, 百拇医药
Key words:Rhizoma Amorphophalli;plant extracts;SGC-7901;cell proliferation;cell cycle;apoptosis
中药蛇六谷是天南星科魔芋属(Amorphophallus)的多年生草本植物的统称，又名魔芋，亦称蒟蒻、蒻头、由跋等。经考证[1-2]，浙江地区药用“蛇六谷”为花魔芋Amorphophallus Konjac K.Koch的块茎。近年来，在上海、浙江地区“蛇六谷”被大量用于肿瘤病的治疗，取得了较好的疗效。鉴于文献报道大多集中于魔芋精粉与魔芋葡甘露聚糖的研究，对原植物蛇六谷抗肿瘤有效成分及其机制的研究较少，本实验拟用系统溶剂法对蛇六谷进行初步提取分离，观察其对体外培养的人胃癌SGC-7901细胞增殖的影响，以分析蛇六谷抗肿瘤的有效部位和作用机理，为临床治疗和新药开发提供理论依据。
1 材料与方法
, 百拇医药
1.1 材料
1.1.1 实验材料 蛇六谷，购自浙江省中医院中药房，产地为浙江临安。经徐锡山主任药师鉴定为天南星科(Araceae)魔芋属花魔芋(Amorphophallus Konjac K.Koch)的干燥块茎。5-氟尿嘧啶/Fluorouracil：江苏南通精华制药有限公司，批号060602。
1.1.2 细胞株 人胃癌细胞株SGC-7901，购自中国科学院上海细胞生物学研究所。
1.1.3 主要试剂 超级新生牛血清(杭州四季青生物工程材料研究所)、RPMI-1640(美国GIBCOBRL公司)、MTT(华美生物工程公司，进口分装)、1∶250胰蛋白酶(吉泰科技有限公司)、二甲基亚砜(分析纯，江苏鸿声化工厂)、碘化丙啶(杭州昊天生物技术有限公司，进口分装)。乙醇、石油醚、乙酸乙酯、石油醚均为分析纯。
1.1.4 主要仪器 SENCO R-201旋转蒸发仪(上海中顺生物科技有限公司)、Heraeus三气细胞培养箱(德国Heraeus公司)、SW-CJ-1F层流超净工作台(苏州安泰空气技术公司)、LD4-2A型低速离心机(北京医用离心机厂)、UR-4100型酶标仪(美国Dynatech公司)、EPICS XL型流式细胞仪(美国COULTER公司)、细胞培养板与培养瓶(丹麦 NUNCLON公司)。, 百拇医药(陈培丰 吴巧凤 丁志山 俞 颖 潘 磊)
关键词：蛇六谷；提取物；人胃癌SGC-7901细胞；细胞增殖；细胞周期；凋亡
, http://www.100md.com
中图分类号：R285.5文献标识码：A
文章编号：1673-7717(2008)11-2313-04
Empirical Study on Effect of Different Extracts of Rhizoma Amorphophalli on the Proliferation of Cell SGC-7901
CHEN Peifeng1,WU Qiaofeng2,DING Zhishan2, YU Ying1, PAN Lei1
(1.The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,Zhejiang,China;
, 百拇医药
2.ZheJiang Chinese Medical University,Hangzhou 310053,Zhejiang,China)
Abstract:Objective:To observe the effect of different extracts of Rhizoma Amorphophalli on the proliferation of cell SGC-7901,and explore the possibility mechanism of them.Methods:Different extracts of Rhizoma Amorphophalli were extracted by the technique of solvent-refining; effect of different dose groups of extracts on cell SGC-7901 were tested by MTT and growth curve method; effects of the extracts of petroleum ether on cell cycle and apoptosis were inspected by flow cytometry.Results:The extracts of ethanol, petroleum ether and ethyl acetate of Rhizoma Amorphophalli have inhibitory effect on the proliferation of cell SGC-7901,and the extract of petroleum ether has much more evident effect, showed a dose-effect curve. Growth curve method showed the extract of petroleum ether and ethyl acetate of Rhizoma Amorphophalli had a time-effect effect on the proliferation of cell SGC-7901. Flow cytometry detection showed the extract of petroleum ether of Rhizoma Amorphophalli can block the cell cycle of SGC-7901 at G0/G1 period,and the mechanism possibly related to inducing apoptosis of tumor cell.Conclusion:The extract of petroleum ether of Rhizoma Amorphophalli is possibly one of antineoplastic parts of the plant.
, 百拇医药
Key words:Rhizoma Amorphophalli;plant extracts;SGC-7901;cell proliferation;cell cycle;apoptosis
中药蛇六谷是天南星科魔芋属(Amorphophallus)的多年生草本植物的统称，又名魔芋，亦称蒟蒻、蒻头、由跋等。经考证[1-2]，浙江地区药用“蛇六谷”为花魔芋Amorphophallus Konjac K.Koch的块茎。近年来，在上海、浙江地区“蛇六谷”被大量用于肿瘤病的治疗，取得了较好的疗效。鉴于文献报道大多集中于魔芋精粉与魔芋葡甘露聚糖的研究，对原植物蛇六谷抗肿瘤有效成分及其机制的研究较少，本实验拟用系统溶剂法对蛇六谷进行初步提取分离，观察其对体外培养的人胃癌SGC-7901细胞增殖的影响，以分析蛇六谷抗肿瘤的有效部位和作用机理，为临床治疗和新药开发提供理论依据。
1 材料与方法
, 百拇医药
1.1 材料
1.1.1 实验材料 蛇六谷，购自浙江省中医院中药房，产地为浙江临安。经徐锡山主任药师鉴定为天南星科(Araceae)魔芋属花魔芋(Amorphophallus Konjac K.Koch)的干燥块茎。5-氟尿嘧啶/Fluorouracil：江苏南通精华制药有限公司，批号060602。
1.1.2 细胞株 人胃癌细胞株SGC-7901，购自中国科学院上海细胞生物学研究所。
1.1.3 主要试剂 超级新生牛血清(杭州四季青生物工程材料研究所)、RPMI-1640(美国GIBCOBRL公司)、MTT(华美生物工程公司，进口分装)、1∶250胰蛋白酶(吉泰科技有限公司)、二甲基亚砜(分析纯，江苏鸿声化工厂)、碘化丙啶(杭州昊天生物技术有限公司，进口分装)。乙醇、石油醚、乙酸乙酯、石油醚均为分析纯。
1.1.4 主要仪器 SENCO R-201旋转蒸发仪(上海中顺生物科技有限公司)、Heraeus三气细胞培养箱(德国Heraeus公司)、SW-CJ-1F层流超净工作台(苏州安泰空气技术公司)、LD4-2A型低速离心机(北京医用离心机厂)、UR-4100型酶标仪(美国Dynatech公司)、EPICS XL型流式细胞仪(美国COULTER公司)、细胞培养板与培养瓶(丹麦 NUNCLON公司)。, 百拇医药(陈培丰 吴巧凤 丁志山 俞 颖 潘 磊)