- 學位論文
紅檜葉化合物對於HT-1080人類纖維肉瘤細胞MMP-2,-9表現之研究
Studies on the Regulation of MMP-2, -9 in HT-080 Human Fibrosarcoma cells by Chemical Constituents of the Leaves of Chamaecyparis formosensis
摘要
紅檜是台灣特有種植物，生長在高海拔地區，屬於長壽植物。這項研究的目的是利用高效液相層儀析結合活性導向(Bioassay-guided)的方法，利用半製備型高效液相層析儀進行紅檜葉乙酸乙酯層之活性成分純化、分離，並結合紅外線光譜、一維及二維核磁共振圖譜與質譜儀確定其結構。利用活性導向(Bioassay-guided)的方法，總共分離出37個化合物(HK1-HK37) ，其中包含4個倍半萜類化合物、13個雙萜類化合物、5個雙黃酮類化合物、4個木質素類化合物、3個黃酮類化合物、4個芳香環類化合物、2個脂肪酸及2個固醇類,上述37個化合物，其中倍半萜類化合物 HK2*、HK3*、HK4*，及雙萜類化合物HK14*為新化合物。 並將所分離出來的成分做MMP-2,-9活性篩選，以HT-1080人類纖維肉瘤細胞，進行凝膠電泳酵素分析方法(Gelatin-Zymograph)， 做MMP-2,-9含量增減的評估，分析純化合物調節情形。發現化合物HK2*、 HK4* 、HK19、HK24、HK25、HK27 及HK28，在不同濃度下，對於MMP-2 ,-9有不同程度的增加。同時也利用MTT assay進行細胞存活率分析，發現活性化合物對HT-1080細胞沒有毒性，不影響所評估的MMP-2,-9之量。故期待從紅檜葉所分離出來的活性天然成分，在未來可做更進一步研究。
並列摘要
Chamaecyparis formosensis Matsum. commonly known as Taiwan red cypress is native to the high mountain area in Taiwan. This species is long-lived. The aims of this study were to use high performance and semi-preparative liquid chromatography combined with Bioassay-guided method to isolate and purify active compounds from Ethyl-acetate layer. Furthermore, using the IR, 1D,2D-NMR, and mass spectrometry to confirm the structures. Bioassay-guided separation led to the identification of 37 compounds from Ethyl-acetate layer, including four sesquiterpenoids，thirteen diterpenoids， five biflavonoids, four phenylpropanoids, three polyphenol compounds,four aromatics,two fatty acid, two steroid， and seven others compounds. Among them, HK2*, HK3*, HK4*, and HK14* are as new compounds by spectroscopic analysis. Compounds separating by Bioassay-guided method were evaluating MMP-2, -9 contents in cultured HT-1080 human fibrosarcoma cells.Compounds HK2*, HK4*, HK19, HK24, HK25, HK27, and HK28 showed the increase in MMP-9 or MMP-2 at different concentrations. In addition, all of them had no cytotoxicity to interference the assessment of MMP-2,-9 evaluating by the MTT cell assay. According to the above results, these bioactivity compounds from C.formosensis will be further studied.