Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22247
標題: Studies on cell growth arrest and anti-inflammatory effects of caffeic acid phenethyl ester and a fermented legume mixture
咖啡酸苯乙酯及豆類發酵液對細胞存活與抗發炎作用之探討
作者: 黃淑琳
Hwang, Shu-Lin
關鍵字: propolis
蜂膠
CAPE
fermented legume mixture
ULS
IC50
nitric oxide
咖啡酸苯乙酯
優豆靈液
毒殺細胞
細胞自噬
一氧化氮
出版社: 分子生物學研究所
引用: 1. Grunberger D, Banerjee R, Eisinger K, Oltz EM, Efros L, Caldwell M, Estevez V, Nakanishi K: Preferential cytotoxicity on tumor cells by caffeic acid phenethyl ester isolated from propolis. Experientia 1988, 44(3):230-232. 2. Chiao C, Carothers AM, Grunberger D, Solomon G, Preston GA, Barrett JC: Apoptosis and altered redox state induced by caffeic acid phenethyl ester (CAPE) in transformed rat fibroblast cells. Cancer Res 1995, 55(16):3576-3583. 3. Chen MF, Wu CT, Chen YJ, Keng PC, Chen WC: Cell killing and radiosensitization by caffeic acid phenethyl ester (CAPE) in lung cancer cells. J Radiat Res (Tokyo) 2004, 45(2):253-260. 4. Chen MJ, Chang WH, Lin CC, Liu CY, Wang TE, Chu CH, Shih SC, Chen YJ: Caffeic acid phenethyl ester induces apoptosis of human pancreatic cancer cells involving caspase and mitochondrial dysfunction. Pancreatology 2008, 8(6):566-576. 5. Jin UH, Song KH, Motomura M, Suzuki I, Gu YH, Kang YJ, Moon TC, Kim CH: Caffeic acid phenethyl ester induces mitochondria-mediated apoptosis in human myeloid leukemia U937 cells. Mol Cell Biochem 2008, 310(1-2):43-48. 6. Bertram JS: The molecular biology of cancer. Mol Aspects Med 2000, 21(6):167-223. 7. Baehrecke EH: Autophagy: dual roles in life and death? Nat Rev Mol Cell Biol 2005, 6(6):505-510. 8. Levine B: Cell biology: autophagy and cancer. Nature 2007, 446(7137):745-747. 9. van der Meer FJ, Faber DJ, Aalders MC, Poot AA, Vermes I, van Leeuwen TG: Apoptosis- and necrosis-induced changes in light attenuation measured by optical coherence tomography. Lasers Med Sci 2010, 25(2):259-267. 10. Okamura T: [Gene, regulation of synthesis, and physiological activity of nNOS]. Nippon Rinsho 2004, 62 Suppl 9:455-459. 11. Nathan C, Xie QW: Nitric oxide synthases: roles, tolls, and controls. Cell 1994, 78(6):915-918. 12. Davis KL, Martin E, Turko IV, Murad F: Novel effects of nitric oxide. Annu Rev Pharmacol Toxicol 2001, 41:203-236. 13. Hofseth LJ: Nitric oxide as a target of complementary and alternative medicines to prevent and treat inflammation and cancer. Cancer Lett 2008, 268(1):10-30. 14. Ohshima H, Bartsch H: Chronic infections and inflammatory processes as cancer risk factors: possible role of nitric oxide in carcinogenesis. Mutat Res 1994, 305(2):253-264. 15. Tsai JT, Liu HC, Chen YH: Suppression of inflammatory mediators by cruciferous vegetable-derived indole-3-carbinol and phenylethyl isothiocyanate in lipopolysaccharide-activated macrophages. Mediators Inflamm 2010, 2010:293642. 16. Srimal RC, Dhawan BN: Pharmacology of diferuloyl methane (curcumin), a non-steroidal anti-inflammatory agent. J Pharm Pharmacol 1973, 25(6):447-452. 17. Chan MM, Huang HI, Fenton MR, Fong D: In vivo inhibition of nitric oxide synthase gene expression by curcumin, a cancer preventive natural product with anti-inflammatory properties. Biochem Pharmacol 1998, 55(12):1955-1962. 18. Ballestrero A, Montemurro F, Gonella R, Capaldi A, Danova M, Friedman D, Puglisi M, Aglietta M, Patrone F: Dose-dense vinorelbine and paclitaxel with granulocyte colony-stimulating factor in metastatic breast cancer patients: anti-tumor activity and peripheral blood progenitor cell mobilization capability. Breast Cancer Res Treat 2003, 82(3):185-190. 19. Wang YY, Khoo KH, Chen ST, Lin CC, Wong CH, Lin CH: Studies on the immuno-modulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucose-containing glycoprotein fraction responsible for the activities. Bioorg Med Chem 2002, 10(4):1057-1062. 20. Ho TF, Ma CJ, Lu CH, Tsai YT, Wei YH, Chang JS, Lai JK, Cheuh PJ, Yeh CT, Tang PC et al: Undecylprodigiosin selectively induces apoptosis in human breast carcinoma cells independent of p53. Toxicol Appl Pharmacol 2007, 225(3):318-328. 21. Lin YH, Chiu JH, Tseng WS, Wong TT, Chiou SH, Yen SH: Antiproliferation and radiosensitization of caffeic acid phenethyl ester on human medulloblastoma cells. Cancer Chemother Pharmacol 2006, 57(4):525-532. 22. Young JE, Martinez RA, La Spada AR: Nutrient deprivation induces neuronal autophagy and implicates reduced insulin signaling in neuroprotective autophagy activation. J Biol Chem 2009, 284(4):2363-2373. 23. Hsieh YH, Chu FH, Wang YS, Chien SC, Chang ST, Shaw JF, Chen CY, Hsiao WW, Kuo YH, Wang SY: Antrocamphin A, an anti-inflammatory principal from the fruiting body of Taiwanofungus camphoratus , and its mechanisms. J Agric Food Chem 2010, 58(5):3153-3158. 24. Su ZZ, Lin J, Grunberger D, Fisher PB: Growth suppression and toxicity induced by caffeic acid phenethyl ester (CAPE) in type 5 adenovirus-transformed rat embryo cells correlate directly with transformation progression. Cancer Res 1994, 54(7):1865-1870. 25. Watabe M, Hishikawa K, Takayanagi A, Shimizu N, Nakaki T: Caffeic acid phenethyl ester induces apoptosis by inhibition of NFkappaB and activation of Fas in human breast cancer MCF-7 cells. J Biol Chem 2004, 279(7):6017-6026. 26. Onori P, DeMorrow S, Gaudio E, Franchitto A, Mancinelli R, Venter J, Kopriva S, Ueno Y, Alvaro D, Savage J et al: Caffeic acid phenethyl ester decreases cholangiocarcinoma growth by inhibition of NF-kappaB and induction of apoptosis. Int J Cancer 2009, 125(3):565-576. 27. Eisenberg-Lerner A, Bialik S, Simon HU, Kimchi A: Life and death partners: apoptosis, autophagy and the cross-talk between them. Cell Death Differ 2009, 16(7):966-975. 28. Yu SH, Kao YT, Wu JY, Huang SH, Huang ST, Lee CM, Cheng KT, Lin CM: Inhibition of AMPK-Associated Autophagy Enhances Caffeic Acid Phenethyl Ester-Induced Cell Death in C6 Glioma Cells. Planta Med 2011.
摘要: 蜂膠中的咖啡酸苯乙酯 (caffeic acid phenethyl ester,CAPE),已被證實具有抗氧化、抗發炎、抗病毒、及抑制癌細胞生長等活性。本研究利用多株癌細胞測試 CAPE 的毒殺細胞作用,發現以終濃度 0-20 μg/mL 的 CAPE 處理癌細胞 HL-60、HepG2、C3A、CL1-0、H460、Caco2、MCF-7、PA-1,48 小時後,細胞之存活率降低,IC50 為 < 2 μg/mL 或介於 4 - 16 μg/mL 之間;但是,以鄉圖濃度的 CAPE 處理正常肺細胞 WI-38 及血球細胞 PBMC,48 小時後,細胞的存活率與控制組相似,IC50皆 > 20 μg/mL。以 15 μg/mL CAPE 處理 CL1-0 細胞 48 小時後,發現於細胞的細胞質有類似細胞自噬體 (Autophagosome) 的囊泡 (vacuole) 出現。利用西方雜配來偵測細胞自噬特徵 LC3-II 蛋白的表現,發現以 15 μg/mL 的 CAPE 處理 CL1-0,隨著處理時間由 0 小時增長到 48 小時,LC3-II 蛋白的表現會上升約 5 倍,表示確有細胞自噬的發生。 本研究亦證明由三合春生技公司的豆類發酵液-優豆靈液 (U-lifespring , ULS) 上清液,能夠抑制老鼠巨噬細胞 Raw 264.7 因 LPS 處理而誘發的一氧化氮 (nitric oxide, NO)的生成,並且此抑制作用為 dose-dependent,也發現 ULS 上清液,能夠使人類肝癌細胞 HepG2死亡。將 ULS 上清液,以等體積的乙酸乙酯 (ethyl acetate,EA)做二次液相-液相劃分,發現二次EA 層具有抗發炎活性及毒殺細胞株 CL1-0、A549、WI-38 的活性,水層則不具有此二活性。進一步利用正向 HPLC 對二次EA 層做分離及純化,發現所分離出來的 13 個分離 fraction ,其中 fraction 10 能夠抑制 Raw 264.7 細胞因 LPS 刺激而產生的一氧化氮達 59%。將 fraction 10 以質譜儀進行身分鑑定,發現 fraction 10 具有一分子量為 345.33 g/mole 的化合物。
Caffeic Acid Phenethyl Ester (CAPE) is found in resinous mixture used by honey bees, commonly called propolis and the CAPE has been shown to have antioxidant, anti-inflammatory, anti-viral and is cytotoxic to cancer cells. The effect of CAPE to kill many strains of cancer cells is studied. It has been found out through experiments that the concentration of 0-20 μg/mL of CAPE to the HL-60, HepG2, C3A, CL1-0, H460, Caco2, MCF-7, PA-1 cancer cell lines showed decreased cell survival after 48 hrs. The Inhibition concentration (IC50) was <2 &micro;g/ml or 4 - 16 &micro;g/mL according to the cell lines. The concentration of CAPE was also used in normal lung WI- 38 cells and normal blood PBMC cells, in 48 hrs, the cell survival and the control group showed >20 &micro;g/mL. CL1-0 cell line was treated with 15 &micro;g/mL of CAPE for 48 hrs. The morphology of the cell was examined using Fluorescence Inverted Microscope (Axiovert 200) and showed vesicles in the cell which may cause in autophagy. Western blotting was done to check the expression of LC3-II protein during autophagy and found out that 15 &micro;g/mL of CAPE from 0 hrs to 48 hrs showed the gradual increase in expression of LC3-II protein and hence the autophagy occurs. This study showed the use fermented legume mixture (U-lifespring, ULS) to examine the anti-inflammatory response against Raw 264.7 macrophage. The ULS supernatant has shown to inhibit the Raw 264.7 mouse macrophages, which contains nitric oxide induced by the LPS treatment. This inhibition is dose dependent manner. The ULS supernatant also showed inhibition against human hepatoma HepG2 cells. Equal volumes of ethyl acetate (EA) were added to the ULS supernatant which is called EA partition. The layer containing the EA has anti-inflammatory activity and showed cytotoxic effect against CL1-0, A549, WI-38 cell lines whereas the water layers do not have any activity. Secondary separation and purification of the EA layer was done using HPLC and isolated 13 separate fractions. The fraction 10 inhibited nitric oxide in the Raw 264.7 cells stimulated by LPS up to 59%. Mass spectrometry was used to identify the fraction 10 and found out that the molecular weight is 353.33 g/mole of the compound.
URI: http://hdl.handle.net/11455/22247
其他識別: U0005-2208201114541700
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2208201114541700
Appears in Collections:分子生物學研究所

文件中的檔案:

取得全文請前往華藝線上圖書館



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.