毛花猕猴桃根活性部位与活性成分制备及其抗肿瘤活性研究

发布时间：2018-06-25 08:13

本文选题：毛花猕猴桃根 + 抗肿瘤活性　；参考：《浙江大学》2015年硕士论文

【摘要】：毛花猕猴桃(Actinidia eriantha Benth.)又称白山毛桃、毛冬瓜,为猕猴桃科猕猴桃属植物。而白山毛桃根是畲族民间治疗肿瘤的习用药材,广泛用于胃癌、乳腺癌、食道癌、鼻咽癌和肝癌等的治疗,但相关的药效物质基础及其作用机制尚不清楚,本研究旨在发现毛花猕猴桃根具有抗肿瘤作用的活性部位及成分,并初步探讨它们的抗肿瘤机制。论文首先采用快速层析法将毛花猕猴桃根的乙醇总提物(AE-E)粗分,得到二氯甲烷洗脱物(AE-E/DM)及甲醇洗脱物(AE-E/M)两个部位；综合采用硅胶色谱柱层析法及葡聚糖凝胶纯化法,分离得到单体化合物(AE-P)；采用质谱及核磁共振技术鉴定AE-P的结构为2α,3α,24-三羟基-12-烯-28-乌苏酸。论文还采用HPLC-PAD法测定了不同批次毛花猕猴桃根药材及不同提取部位中2α,3α,24-三羟基-12-烯-28-乌苏酸的含量。论文采用MTT法对毛花猕猴桃根的不同提取、分离部位及化合物2α,3α,24-三羟基-12-烯-28-乌苏酸进行了活性筛选,结果表明AE-E/DM对人肝癌细胞株HepG2,人前列腺癌细胞株pc3,人胃癌细胞株BGC-823,人结直肠腺癌sw620的增殖均有显著地抑制作用,推测其可能为毛花猕猴桃根发挥抗肿瘤作用的潜在活性部位；2α,3α,24-三羟基-12-烯-28-乌苏酸亦能显著地抑制以上四种癌细胞增殖。论文采用人结直肠腺癌sw620裸鼠移植瘤模型对2α,3α,24-三羟基-12-烯-28-乌苏酸的体内抗肿瘤活性进行了验证,证实其能剂量依赖性的抑制人结直肠腺肿瘤的生长,推测其可能为毛花猕猴桃根发挥抗肿瘤作用的潜在活性成分之一。论文中进一步采用AO/EB染色、流式细胞术以及western blot技术初步探讨了2α,3α,24-三羟基-12-烯-28-乌苏酸抗结直肠腺癌的作用机制。结果表明其能引起细胞凋亡,并伴随自噬。本研究为揭示毛花猕猴桃根的药效物质基础奠定了一定的基础,为其进一步在抗肿瘤领域的应用提供了初步的科学数据。
[Abstract]:(Actinidia eriantha Benth. of Actinidia chinensis L. It is also called Baishan Maotao, the white winter melon, for the Actinidia family Actinidia plants. Peach root in Baishan Mountain is a folk medicine used in the treatment of cancer of she nationality. It is widely used in the treatment of gastric cancer, breast cancer, esophagus cancer, nasopharyngeal carcinoma and liver cancer. However, the relevant pharmacodynamic material basis and its mechanism of action are still unclear. The aim of this study was to find out the active parts and components of Actinidia deliciosa root with anti-tumor effect and to explore their anti-tumor mechanism. In this paper, the ethanol total extract (AE-E) of Actinidia chinensis root was extracted by rapid chromatography, and two parts of AE-E / M (AE-E / DM) and AE-E / M (AE-E / M) were obtained, and the silica gel chromatographic column chromatography and dextran gel purification were used. The monomer compound (AE-P) was isolated, and the structure of AE-P was identified by mass spectrometry and nuclear magnetic resonance (NMR). HPLC-pad method was also used to determine the content of 2 伪 -trihydroxy-12ene-28-ursolic acid in different batches of Actinidia chinensis root and its extracts. In this paper, MTT method was used to screen the different extraction, isolation site and activity of 2 伪 -trihydroxy-12-ene-28-ursolic acid from Actinidia chinensis root. The results showed that AE-E / DM could significantly inhibit the proliferation of human hepatoma cell line HepG2, human prostate cancer cell line pc3, human gastric cancer cell line BGC-823 and human colorectal adenocarcinoma sw620. The proliferation of these four kinds of cancer cells was also significantly inhibited by 2 伪 -trihydroxy-12en-28-ursolic acid. The in vivo antitumor activity of 2 伪 -3 伪 -trihydroxy-12-ene-28-ursolic acid was tested in nude mice with human colorectal adenocarcinoma (sw620). It was proved that it could inhibit the growth of human colorectal adenoma in a dose-dependent manner. It is speculated that it may be one of the potential active components of Actinidia chinensis root to play an anti-tumor effect. In this paper, AO-EB staining, flow cytometry and western blot technique were used to investigate the mechanism of anti-colorectal adenocarcinoma. The results showed that it could induce apoptosis, accompanied by autophagy. This study has laid a certain foundation for revealing the substance basis of medicinal effect of Actinidia chinensis root, and provided preliminary scientific data for its further application in the field of anti-tumor.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S663.4

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