摘 要：目的 通过观察泡核桃青皮提取物1,2,4,6-Tetragalloyl glucose对结肠癌细胞存活率的影响, 为1,2,4,6-Tetragalloyl glucose治疗结肠癌提供一定的理论依据，以探讨其对人类结肠癌药物治疗的机制。 方法 体外培养肿瘤细胞株HT-29,A549, PC3, HePG2 和人正常细胞株HEK293，从泡核桃青皮中分馏、萃取和纯化出1,2,4,6-没食子酰葡萄糖（1,2,4,6-Tetragalloyl glucose），采用不同浓度的1,2,4,6-Tetragalloyl glucose（3.125, 6.25, 12.5, 25, 50, 100 g/ml）对结肠癌细胞株和人正常细胞进行干预，用四甲基偶氮唑盐(MTT)比色法检测其对人结肠癌细胞株存活率的影响。结果 MTT法检测1,2,4,6-Tetragalloyl glucose对结肠癌细胞株存活率的具有抑制作用，且对HT29细胞株抑制最为明显。结论  从泡核桃青皮中提取的1,2,4,6-Tetragalloyl glucose对体外培养的结肠癌细胞存活率有明显抑制作用，具体作用机制有待进一步研究。54348

毕业论文关键词： 1,2,4,6-Tetragalloyl glucose，结肠癌细胞，MTT法，萃取

Abstract：Objective  It provided a theoretical basis for 1,2,4,6-Tetragalloyl glucose treatment of colon cancer by observing the its effect on the viability of colon cancer cells and to investigate the mechanisms of human colon cancer drug therapy. Methods  We culture colon cancer cell lines HT-29, A549, PC3, HePG2 (full name,except HT-29, other cell lines were not human colon cancer cell, please  CHECK IT) and normal human cell lines HEK293. Extraction, fractionation, and purification of galloyltannins from seed husks of J. sigillata was carried out under activity-guide way. The cytotoxicity of different concentrations of 1,2,4,6-Tetragalloyl glucose (3.125, 6.25, 12.5, 25, 50, 100 g / ml) were investigated by  tetrazolium salt (MTT) assay, moreover, the apoptic rate was investigated by flow cytometry. Result  1,2,4,6-Tetragalloyl glucose inhibited the cell viability of colon cancer cell lines in a concentration-dependent manner. Conclusion  1,2,4,6-Tetragalloyl glucose significantly inhibited colon cancer cell survival  in vitro.  Mechanism needs further study in detail.

Keywords：1,2,4,6-Tetragalloyl glucose, Colon cancer cells, MTT assay, apoptosis

目   录

1 前言 2

1.1 泡核桃简介 2

1.1.1泡核桃功效 3

1.2 关于结肠癌的认识 4

1.2.1结肠癌的发病机理 4

1.2.2 结肠癌的治疗 4

2  材料与方法 5

2.1 材料 5

2.1.1 实验材料 5

2.1.2 实验主要试剂 5

2.1.3 实验主要仪器 5

2.2 方法 5

2.2.1 原材料的分馏萃取 5

2.2.2结肠癌细胞培养 6

2.2.3 MTT法检测粗提取物B1~B6对结肠癌细胞株增殖的抑制作用 6

2.2.4 MTT法检测 1,2,4,6-Tetragalloyl glucose对结肠癌细胞增殖的抑制作用 8

2.2.5 不同浓度1,2,4,6-Tetragalloyl glucose对HT-29细胞活性的影响 8

2.2.6 Annexin V-FITC染色法检测细胞凋亡率 8

3 结果与分析 8

3.1检测粗提取物B1~B6对结肠癌细胞株增殖的抑制作用 8

3.2 1,2,4,6-Tetragalloyl glucose对结肠癌细胞增殖的抑制作用