PDF《Study on liver protective effect》

保肝;

活性成分;

药理机制;

泡腾片. 澳门大学硕士学位论文 III Abstract Study on liver protective effect and pharmaceutical design of Penthorum chinense Pursh Yangyang Hu Supervisor: Prof. Yitao Wang Institute of Chinese Medical Sciences Penthorum Chinense Pursh, a traditional Chinese medicine derived from Chinese ethnic group Miao, has been used to protect liver from damaging by alcohol and toxins for a long history. Modern research have showed that Penthorum Chinense Pursh containing many active ingredients such as alkaloids, flavonoids, glucose, and so on. It has a variety of activities, including detoxification, activating blood and dissolving stasis, liver-protecting, tonifying spleen, as well as anti-virus. However, until now, the liver protective effect of Penthorum Chinense Pursh remains blurry. In order to broaden the application of Penthorum Chinense Pursh, this study was designed to identify its active constitutes and explore the mechanism in protecting liver damage. In order to obtain stable samples, the chemical constitutes and extraction process were studied. Based on the in vitro t-BHP/ethanol-induced liver damage model, I found that the water extract of Penthorum Chinense Pursh could significantly protect HL-7702 cells from t-BHP induced damage, and the mechanism may be associated with its ability of scavenging ROS generation. In conclusion, with the aim to provide scientific foundation for further development and application of Penthorum Chinense Pursh, this study partially clarified its chemical constitutes and pharmacological mechanism, as well as a preliminary formulation for developing effervescent tablets. This study may benefit the further application of Penthorum Chinense Pursh in clinic. 澳门大学硕士学位论文 IV Key words Penthorum Chinense Pursh, chemical constitutes,pharmacological machanism, effervescent tablets. 澳门大学硕士学位论文 V 目录致谢I摘要II Abstract.III 目录.V

图表目录.VII

第一章 前言.1 1.1 赶黄草概述.1 1.2 赶黄草的生药学研究.1 1.2.1 赶黄草形态特徵.1 1.2.2 赶黄草叶片显微鉴别.2 1.2.3 赶黄草茎显微特徵.2 1.3 赶黄草的化学研究.3 1.3.1 赶黄草化学成分研究.3 1.3.2 提取工艺研究.5 1.4 赶黄草的药理作用.6 1.4.1 利胆、退黄及降酶作用.6 1.4.2 保肝作用.6 1.4.3 抗病毒作用.7 1.4.4 抗氧化作用.8 1.4.5 抗突变作用.8 1.5 展望.8

第二章 赶黄草的提取工艺研究.10 2.1 试验材料.10 2.1.1 仪器.10 2.1.2 试剂与药材.10 2.2 试验方法及结果.10 2.2.1 提取条件考察.10 2.2.2 赶黄草提取物化学成分谱研究.11 2.2.3 赶黄草水提物化学成分研究.13 2.4 小结.13

第三章 赶黄草保肝作用体外研究.14 3.1 试验材料.14 3.1.1 仪器.14 3.1.2 试剂和药品.14 3.2 细胞培养和传代.15 3.3 肝损伤体外模型的建立.15 3.3.1 过氧化氢诱导的肝损伤模型的建立.15 3.3.2 乙醇诱导的肝损伤模型的建立.16 澳门大学硕士学位论文 VI 3.4 MTT 法检测细胞存活率

17 3.4.1 赶黄草体外肝毒性的筛选.17 3.4.2 赶黄草体外保肝作用的筛选.18 3.5 Hochest 染色法检测细胞凋亡.20 3.6 JC-1 染色法检测细胞膜电位

22 3.7 细胞内 ROS 的检测.23 3.7.1 萤光显微镜观察细胞内 ROS.23 3.7.2 细胞内 ROS 的定量检测.24 3.7.3 流式细胞仪检测细胞内 ROS.25 3.7.4 肝细胞内 ROS 检测结果分析.26 3.7 WB 法检测凋亡相关蛋白的表达

27 3.8 小结.29

第四章 赶黄草泡腾片的处方优化及制备工艺研究.30 4.1 实验材料.30 4.1.1 实验仪器.30 4.1.2 试剂和药品.30 4.2 实验方法.30 4.2.1 pH 值的测定方法.30 4.2.2 崩解时限的测量方法.30 4.2.3 颗粒休止角测定.30 4.2.4 成型性实验.31 4.2.5 吸湿性实验.31 4.2.6 临界相对湿度(CRH)的测定.31 4.3 辅料的筛选.31 4.3.1 填充剂的筛选.31 4.3.2 润滑剂的选择.32 4.3.3 黏合剂的选择.32 4.3.4 泡腾剂的选择.32 4.3.5 正交试验优化辅料组成.33 4.4 制备处方及工艺.34 4.4.1 处方.34 4.4.2 制法.34 4.5 小结.34