民族药地稔抗妇科恶性肿瘤药效物质基础研究

The national medicine Melastoma dodecandrum Lour. has the functions of detoxification, cooling and hemostasis. It is a good medicine for the treatment of gynecological diseases. Based on the analysis of the quality of the Melastoma dodecandrum Lour. and its preliminary anti-cancer screening,Based on previous research, MTT assay or CCK-8 method will be used to investigate the anti-proliferative effects of extracts, different extracts and components of Melastoma dodecandrum Lour. as the research object, on human ovarian cancer cell lines SK-OV-3, HO-8910, human cervical cancer cells HeLa, SiHa, human endometrial cancer cell lines Hec-1B, HHUA, etc,which could screen out the site of anti-tumor activity. Single factor experiments and response surface methodology will be used to optimize the extraction of polyphenols in Melastoma dodecandrum Lour. and the anti-tumor active components were purified by modern methods. We will analyze the anti-tumor active site by high performance liquid chromatography (HPLC) combined with ELSD or MS.The further evaluation of the antitumor effect of the active site will be achieved through a mouse tumor transplant model. And then, BP neural network and grey relational degree model will be used to analyze the relationship between the active site fingerprints and efficacy results and find out the effective anti-tumor activity components and main components of Melastoma dodecandrum Lour., which is to explore the correlation between changes in chemical composition and anti-tumor activity in fingerprints ,so that clarify the material basis of anti-tumor in Melastoma dodecandrum Lour.

民族药地稔具有清热解毒、活血止血的功效，是疗效确切的妇科良药。在前期地稔质量分析和初步抗癌筛选研究的基础上，以地稔不同提取物和组分为研究对象，采用MTT法或CCK-8法考察地稔提取物对体外培养人卵巢癌细胞株SK-OV-3、HO-8910，人宫颈癌细胞Hela、SiHa,人子宫内膜癌细胞株Hec-1B、HHUA等肿瘤细胞的抗增殖作用，筛选出地稔抗肿瘤活性部位;采用单因素试验结合响应面法优化提取地稔中多酚类组分，以现代分离方法纯化地稔抗肿瘤活性组分；建立高效液相色谱（HPLC）法联合ELSD或MS对地稔抗肿瘤活性部位进行分析；通过小鼠肿瘤移植性模型进一步评价地稔活性部位抗妇科恶性肿瘤作用；采用BP神经网络结合灰色关联度模型分析地稔的活性部位指纹图谱与药效结果之间的联系，寻找地稔中抗肿瘤活性有效组分群及主要成分，探讨化学成分的变化与抗肿瘤活性的相关性，阐明地稔抗肿瘤的物质基础。

民族药地稔为野牡丹科植物地菍的干燥全草，是一种疗效确切的妇科良药。前期研究发现，地稔提取物具有潜在的抗肿瘤活性，但存在药效物质基础不清、作用机制不明确和检测困难等问题，无法为其进一步的临床应用提供科学依据。为了探明地稔提取物的药效物质基础和抗肿瘤作用机制，本项目根据地稔的药性及用药特点，以人宫颈癌、人子宫内膜癌、人乳腺癌和人卵巢癌等妇科肿瘤为研究对象，采用现代药理学和药物分析方法对地稔提取物开展相关基础研究。结果显示，地稔总多酚为抑制人宫颈癌细胞的活性部位，且对其提取及纯化工艺进行优化后，可显著提高地稔总多酚抗肿瘤活性；建立了地稔总多酚UPLC指纹图谱，并通过超高效液相色谱-四极杆飞行时间质谱（UHPLC-QTOF-MS）技术确证了地稔总多酚指纹图谱的共有峰，其中没食子酸、鞣花酸、原儿茶酸等成分与宫颈癌SiHa细胞IC50值呈负相关，同时建立了一测多评法分析地稔质量，为地稔质量标准提升提供了科学依据；地稔总多酚能有效抑制宫颈癌，促进SiHa和HeLa细胞调亡，并通过影响宫颈癌细胞周期的分布，将细胞周期阻滞到GO/G1期，显著抑制宫颈癌细胞的迁移；与正常组相比，地稔总多酚高、中、低剂量组均具有良好的SiHa肿瘤抑制作用，并能显著延长荷宫颈癌SiHa肿瘤小鼠的存活时间。本项目为民族药地稔的深入研究和应用开发提供了实验基础和理论依据。