【作者】 田莉；

【导师】 周秋丽； 房学迅；

【作者基本信息】 吉林大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 基质金属蛋白酶(MMPs)是一类Zn2+依赖的内肽酶家族,在正常和病理情况下的细胞外基质的重塑过程中发挥重要的作用。MMPs在调控细胞因子、生长因子、激素和细胞粘附分子受体等的合成和分泌中发挥重要作用,进而参与细胞凋亡、形态发生、增殖和发育等一系列病理和生理过程。当MMPs的调控机制失去平衡,MMPs活性异常升高时,就会导致一些病理过程的发生。因此,以MMPs为治疗靶点,寻找MMPs的拮抗药物,尤其是MMPs选择性抑制药物,已经成为筛选抗肿瘤药物的热点。本论文通过从中药连翘中分离提取出能够抑制MMPs活性的单体化合物连翘酯苷。通过酶学方法和分子对接模拟方法检测其对MMPs活性的抑制作用,并从通过起其对细胞行为学的影响检测其在细胞学水平的影响,发现连翘酯苷对MMPs的抑制作用具有选择性,这将为新型的MMPs特异性抑制剂的开发提供思路。更多还原

【Abstract】 The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidase involved in the degradation and remodeling the extracellular matrix proteins. The activities of these enzymes are well regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs). Chronic stimulation of MMPs activities due to an imbalance in the levels of MMPs and TIMPs has been implicated in the pathogenesis of a variety of diseases such as cancer, osteoarthritis, rheumatoid arthritis. The MMPs which have been proved top lay key roles in the processes of tumor growth, invasion, metastasis and angiogenesis,are frequently overexpressed in malignant tumors, and have been associated with an aggressive malignant phenotype and poor prognosis inpatients with cancer. Thus it is believed that the growth and metastasis can be controlled by MMP inhibitors (MMPIs) which are expected to be useful for the treatment of cancer. Nowadays, many of the inhibitors of matrix metalloproteinase have come into clinical trial, but to most of our disappointment, none was tested to be a candidate for further curing disease. The main problem of this is the weak selectivity of MMPIs, because a long time non-selectivity inhibition of MMPs may bring out some unexpected biological disorder. In conclusion, it is foremost meaningful to find a MMPI with a high selectivity, affinity and biological availability.Forsythiaside belonging to the derivatives of caffeinic acid is the effective ingredient of the Chinese herb Forsythia. In this thesis, Forsythiaside has been obtained by using Macro-reticular resin D101 and HPLC, which have some obvious advantages: saving solve, time and having high performance. The inhibition ability to MMPs of forsythiaside was tested by monitoring the strength of fluorescent emissioning from the cleaved substrate of MMPs. Then we docked the forsythiaside to different MMPs providing a detailed inhibition mechanism. At last, we observed the affect of forsythiaside to HT1080 cells on the aspect of cell proliferation, 3-D culture, migration and invasion. All the results showed that forsythiaside has higher inhibition ability to MMP-13 than others with a dose-dependent way which is consistent to data of docking. The docking as well suggested that forsythiaside located the S1 and S1’pocket of MMPs. Forsythiaside can inhibit the migration and invasion of HT1080 cells in vitro. The specific mechanism to be further verified. As a result, the specific MMP inhibition of forsythiaside will not only greatly help us to find a leading more potent compound but provide a new strategy for MMPIs development.We can continue studying in the future, it is investigated to pay attention to the inhibition of this new MMPI, at the same time, we should take attention to activities, toxicities, pharmacology, biological, pharmacokinetic and drug metabolism of forsythiaside further. On one hand facilitate our synthesizing and finding new MMP inhibitor, thus develop and regard MMP as the molecule target new medicine with independent intellectual property right; On the other hand explain the function mechanism of traditional Chinese medicine have benefits greatly from molecule level for us, and has offered basis for developing, synthesizing and transforming the medicine guide compound and guidance. Continue launching animal’s model experiment and clinical experiment in a situation that a large number of external experiments are ripe, the mechanism of treating tumour for traditional Chinese medicine studies opens up the new broad space.更多还原