臺灣博碩士論文加值系統

本文將針對台灣綠蜂膠最高等級 (TW-I) 的活性成分、生物活性、膠源植物、與蜜蜂採集台灣綠蜂膠的行為做一系列探討。台灣綠蜂膠的主要含有蜂膠素C、D、F與G並且最具代表性，本文首先以不同溶劑 (甲醇、乙醚、不同百分比乙醇與水) 萃取台灣綠蜂膠，蜂膠塊以99.5%、95%乙醇進行萃取其產率達66.75 ± 0.5% 與66.25 ± 0.5%最佳，而以水萃取僅得7.00±0.82%的產率最差，其他溶劑萃取則在這兩者中間；液相層析儀 (HPLC) 分析各樣本蜂膠素含量後，蜂膠素C、D、F與G總量以99.5%與95%乙醇的萃取液分別含36.73 ± 0.80及37.55 ± 1.29 mg/mL 為最多，水萃取蜂膠含少量蜂膠素而無法測得。生物活性方面進行清除DPPH自由基活性與抗菌活性的分析。不同溶劑萃取的蜂膠中以70%乙醇萃取蜂膠清除DPPH自由基的活性最佳，IC50為16.09 ± 0.13 µg/mL，而水萃取蜂膠的IC50為126.43 ± 2.47 µg/mL，效果最差。4種蜂膠素清除DPPH自由基的效力為C＞D＞G＞F。另外對各溶劑蜂膠萃取液進行抗菌能力分析發現各萃取液對Staphylococcus aureus的最低抑菌濃度與最低殺菌濃度平均為20 g/mL，唯水萃蜂膠無抑菌能力。若分別探討各蜂膠素C、D、F與G對6株革蘭氏陽性菌(Staphylococcus aureua, 3 strains、Bacillus subtillis、Listeria monocytogenes、Paenibacillus larvae)與2株革蘭氏陰性菌 (Escherichia coli、Pseudomonas aeruginosa)的抑菌力，結果顯示4種蜂膠素皆能有效抑制革蘭氏陽性菌，其中尤以蜂膠素C的抑菌力最佳，其對P. larvae的最低抑菌濃度僅2.5 g/mL，最低殺菌濃度則為5.0 g/mL；但4種蜂膠素對於E. coli與P. aeruginosa抑菌效果不佳 (MIC>640 g/mL)。協力作用則以Propolin2C+D的協力效果最好，最低的抑菌濃度與殺菌濃度分別為0.3125 g/mL與0.625g/mL，效果可以比蜂膠素C單獨存在時強2-8倍，但是仍對革蘭氏陰性菌抗菌效果不彰。如此顯示台灣蜂膠分離的蜂膠素對革蘭氏陽性菌具有優異的抑菌與殺菌力。膠源植物的確認則是根據彥臣公司的研究資訊得知血桐果實是台灣蜂膠的膠源植物，因此採集不同成熟度之血桐果實進行研究，以甲醇為溶劑萃取血桐果實表面物質，經HPLC分析後得到與台灣綠蜂膠成分標準品相符合之液相層析圖譜。而血桐果實表面物質萃取液也具有清除DPPH自由基的效果，對S. aureus 同樣具有良好抑菌效果，這個結果更證實血桐果實確實是台灣綠蜂膠之膠源植物。為探討蜜蜂的蜂膠採集行為，於2007年7月在嘉義縣梅山鄉彥臣蜂場選取6群蜜蜂，於每日8:00-18:00連續3日放置花粉收集器以收集全日採集的採集物，並以液相層析儀分析各樣品蜂膠素的含量，用以探討蜜蜂採膠行為。結果顯示，試驗蜂群的蜂膠採集主要分布於8:00-14:00，蜂膠採集量約佔總採集量的77%，而14:00-18:00的採集量僅佔23%，如此可以推測西洋蜂採集綠蜂膠的行為主要於14:00前，14:00以後則蜂膠採集量大幅減少。

At present study, we focused on the compounds, bioactivities, plant origin and the collecting behavior of honey bee of the greatest grade of Taiwanese green propolis (TW-I). The main and also the most representative active compounds of Taiwanese green propolis are propolin C, D, F and G. The first, Taiwanese green propolis were extracted with different solvents (methanol, ethyl easter, ethanol in different percentage, water). The maximum dry matter yield were 66.75 ± 0.5% and 66.25 ± 0.5% extracted with 99.5% and 95% ethanol, and the worst dry matter yield of 7.00±0.82% were extracted by water, others were between these samples. After analysing the amount of propolins, the maximum total amount of C, D, F, G were 36.73 ± 0.80 and 37.55 ± 1.29 mg/L that extracted with 99.5% and 95% ethanol. The amounts of propolins in water extract propolis were unable to be examed. DPPH scavenging and antibacterial activities were carried out in bioactivity test. The best IC50 of DPPH scavenging were 16.09 ± 0.13µg/ml, extracted with 70% ethanol. The IC50 of water extracted propolis was 126.43 ± 2.47 µg/ml, the worst. DPPH scavenging efficacy of the four propolins is ranked as C＞D＞G＞F。Additionally, we evaluated the antibacterial efficacy of each extraction, and the MIC and MBC of the extractions were averagely 20 g/mL, no antibacterial activity was found in the water extracted propolis. Survey of the antibacterial efficacy of propolins C, D, F, and G respectively against 6 gram-positive strains (Staphylococcus aureua, 3 strains, Bacillus subtillis, Listeria monocytogenes, and Paenibacillus larvae) and 2 gram-negative strains (Escherichia coli, Pseudomonas aeruginosa). It was effective for four propolins against gram-positive strains. Propolin C was especially the most effective, the MIC and MBC was only 2.5 g/mL and 5.0 g/mL respectively against P. larvae. The antibacterial efficacy of the propolins against gram-negative strains were not well (MIC>640 g/mL). Propolin2C+D is the most effective synergistic agent, and the minimum bacteriostatic and bactericidal concentration were respectively 0.3125 g/mL and 0.625g/mL. Efficacy of the agent is 2-8 times stronger than propolin C exist singly, but no efficiency against gram negative bacteria. The results demonstrated that propolins isolated from Taiwanese propolis have excellent antibacterial activity and mainly against gram-positive strains. According to the research information from NBM company knew that the plant origin of Taiwanese green propolis is Macaranga tanarius, and different maturity of Macaranga tanarius fruits were collected and analysied. The surface substance of Macaranga tanarius were extracted with methanol. After analysis, the HPLC profiles of the surface substance extraction of M. tanarius were correspondence with the standard of propolis components. Also the surface substance extraction of M. tanarius shows well DPPH scavenging efficacy and antibacterial effect against S. aureus. For investigating the collecting behavior of honey bee, we collected 6 groups of honey bee by pollen collector from 8:00-18:00, 3 days continuous in apiary of NBM company in Mei-Shan in July, 2007. Propolins in samples were analyzed by HPLC to study the collecting behavior of honey bee. The collection of tested group was main at time period of 8:00-14:00, the amount occupied 77% of total amount of propolis collection, and the amount at time period of 14:00-18:00 was only occupied 23% of total amount of propolis collection. The results indicate that most of the propolis collecting behavior of Apis mellifera occurs before 14:00, and the amount of collection decrease substantially after 14:00.

壹、緒言………………………………………………………………………………….1
貳、往昔研究…………………………………………………………………………….3
一、蜂膠……………………………………………………………………………….3
二、蜂膠的性質與組成……………………………………………………………….3
三、蜂膠的生物活性與藥理效用………………………………………………….....4
1. 抗微生物活性...................................................................................................5
A. 抗細菌與真菌活性………………………………………………………….5
B. 抗病毒、原蟲與寄生蟲活性……………………………………………..….6
2. 肝臟的保護作用……………………………………………………………...6
3. 抗氧化活性…………………………………………………………………...7
4. 抗腫瘤活性…………………………………………………………………...7
5. 抗發炎活性…………………………………………………………………...8
6. 其他活性……………………………………………………………………...8
四、台灣綠蜂膠……………………………………………………………….............9
1. 台灣蜂膠的開發…………………………………………………………........9
2. 台灣綠蜂膠與蜂膠素…………………………………………………………9
3. 台灣蜂膠的生物活性………………………………………………………..10
A. 抗微生物活性……………………………………………………………...10
B. 抗氧化活性………………………………………………………………...11
C. 細胞毒殺活性……………………………………………………………...12
五、血桐……………………………………………………………………….......…13
1. 血桐簡介……………………………………………………………………..13
2. 由血桐分離出之化學成分及其功效……………………………………......13
3. 血桐與蜂膠…………………………………………………………………..14
六、蜜蜂的採膠行為………………………………………………………...............14
參、材料與方法………………………………………………………………………...15
一、 樣本萃取………………………………………………………………………15
1. 不同溶劑萃取蜂膠………………………………………………………......15
2. 血桐果實的萃取…...………………………………………………………...15
二、 萃取物的分析………………………………………………………….............15
1. 不同溶劑的萃取效果………………………………………………….….…15
2. 血桐……………………………………………………………………..……16
三、 抗微生物活性.………………………………...……………………………….16
1. 微生物來源與培養條件……………………………………………………..16
2. 不同溶劑蜂膠萃取物的抗菌效果……………………………………….….16
3. 不同蜂膠素的抗菌活性……………………………………………………..17
4. 蜂膠素的協力抗菌活性………………………………………….………….17
5. 血桐…………………………………………………………………………..17
6. MIC與MBC的判定...……………………………………………………….17
四、 清除DPPH自由基效力………………………………………………….……18
1. 不同溶劑蜂膠萃取物…………………………………………………….….18
2. 蜂膠素樣本處理……………………………………………………………..18
3. 血桐果實表面物質萃取物樣本處理………………………………………..18
4. 清除DPPH自由基效力實驗步驟…………………………………………..18
五、 蜜蜂的採膠行為.…...……………………………………………………….…18
1. 樣本的收集與分析…………………………………………………………..18
2. 採集量分析………………………………………………………………..…19
肆、結果…………………………………………………………………………….…..20
一、萃取物的分析……………………………………………………………...……20
1. 不同溶劑的萃取產率………………………………………………………..20
2. 不同萃取液的蜂膠素含量…………………………………………………..20
3. 血桐果實的萃取物分析……………………………………………………..20
二、蜂膠抗微生物活性………...……………………………………………………21
1. 不同溶劑萃取物的抗微生物活性…………………………………………..21
2. 蜂膠素的抗微生物活性與協力作用………………………………………..21
3. 血桐的抗微生物活性………………………………………………………..23
三、 清除DPPH自由基效力……………………………………………………….23
1. 不同溶劑萃取物的DPPH自由基清除效力………………………………..23
2. 蜂膠素的DPPH自由基清除效力…………………………………………..23
3. 血桐果皮物質對DPPH自由基的清除效力………………………………..24
四、 蜜蜂的採膠行為……………………………………………………………….24
伍、討論………………………………………………………………………………...26
一、不同溶劑的萃取效果…………………………………………………...………26
二、膠源植物………………………………………………………………………...26
三、蜂膠素的生物活性……………………………………………………...………28
四、蜜蜂的採膠行為………………………………………………………...………29
陸、參考文獻…………………………………………………………………………...31
柒、圖表………………………………………………………………………………...44
捌、附錄………………………………………………………………………………...64







表次
表一、 以不同溶劑萃取台灣綠蜂膠之乾物質產率(%)與蜂膠素含量(mg/mL)……44
表二、血桐果實表面物質萃取液的乾物質產率(%)與蜂膠素含量(mg/mL)………..45
表三、不同萃取液對金黃葡萄球菌與大腸桿菌的最低抑制與殺菌濃度(µg/mL)….46
表四、不同蜂膠素與全蜂膠萃取液的最低抑菌濃度(µg/mL)……………………….47
表五、不同蜂膠素與全蜂膠萃取液的最低殺菌濃度(µg/mL)……………………….48
表六、蜂膠素抗菌協力作用的最低抑菌濃度(µg/mL)……………………………….49
表七、蜂膠素抗菌協力作用的最低殺菌濃度(µg/mL)……………………………….50
表八、血桐果實表面萃取物的抗微生物活性………………………………………...51
表九、不同台灣綠蜂膠萃取物對清除DPPH自由基的半抑制劑量( IC50) …………52
表十、不同蜂膠素對清除DPPH自由基的半抑制劑量( IC50) ………………………53
表十一、血桐果實表面物質萃取物表對清除DPPH自由基的半抑制劑量 ( IC50)..54
表十二、調查期間試驗蜂群的每日平均採集蜂膠與花粉量 (g/colony/day)……….55
表十三、不同蜂群採集蜂膠與花粉的日平均量………………………………….. …56
表十四、試驗蜂群於每日不同時間點的2小時蜂膠與花粉採集量…………………57
附表一、台灣產蜂膠的分級與組成………………………………………………...…64









圖次
圖一、蜂膠素的HPLC層析圖譜……………………………………………………...58
圖二、蜂膠素抗菌協力作用的最低抑菌濃度 (µg/mL)……………………………...59
圖三、蜂膠素抗菌協力作用的最低殺菌濃度 (µg/mL)……………………………...60
圖四、蜂膠素與咖啡酸苯乙酯對DPPH自由基的清除能力………………………...61
圖五、血桐果實表面物質萃取液對自由基的清除能力……………………………...62
圖六、各蜂群採粉與採膠的趨勢……………………………………………………...63
附圖一、(A) 血桐植株；(B) 血桐果實..………………………………………………65
附圖二、(A) 蜜蜂採集血桐果實表面物質；(B) 左: 蜜蜂花粉藍中的膠塊；右: 蜜蜂採集後血桐果實表面剩餘物質……………………………………………..…..….66
附圖三、血桐嫩果 (右) 與熟果 (左)……………………………………...…………67

山本修。1997。蜂膠--又稱為蜂脂的植物性樹脂。中華健康雜誌 16: 16-23。
川村賢司。1996。蜂膠之有效成分探討。蜂膠健康讀本 1: 22-24。
江文德。1998。簡介大豆中的異黃酮素。食品工業 30: 6-12。
佚名。2003。行政院衛生署中醫藥委員會。臺灣藥用植物資源名錄，血桐。289頁。
朱燕華。1998。類黃酮之介紹。食品工業 30: 1-5。
巫雪呅。2001。台灣產蜂膠的組成與生物活性。國立台灣大學昆蟲學研究所 碩士論文。
李榮明、陳宗欣及李秉鈺。1995。蜂膠臨床應用之展望。藥學雜誌 11: 103-108。
林怡君。2007。以酵素法產製之幾丁寡醣特性及其抗菌活性之研究。國立宜蘭大學食品科學系 碩士論文。
侯加強。1993。如何提高蜂膠酊的質量。養蜂科技 6: 21。
施介人。1998。簡介槲皮酮。食品工業 30: 53-61。
徐玲云、袁譯良。1989。蜂膠質量標準的探討。中國養蜂 2: 6-8。
唐傳和、孟岳成。1999。蜂膠的生理功能以及開發研究。食品工業科技 20: 30-32。
陳裕文。1997。幼蟲芽孢桿菌對蜜蜂的致病力與防治法。國立台灣大學植物病蟲害學研究所 博士論文。
陳裕文、陳嘉南、林仁混及何鎧光。2006。台灣蜂膠的研究與開發。台灣昆蟲特刊 8: 51-65。
陳嘉南。2003。台灣蜂膠之抗癌機制與促進神經幹細胞生長及分化的探討及茶成分對神經幹細胞分化之研究。國立台灣大學生物化學暨分子生物研究所 博士論文。
陳山禾。2006。利用chitin之去乙醯度控制酵素水解產物大小及其抑菌活性之探討。國立東華大學生物技術研究所 碩士論文。
章景瑞。1996。蜂膠王漿和干擾素對純皰疹病毒的作用。蜜蜂 3: 21-22。
解玉軫。1999。不同種類蜂膠組成分分析及抗生特性研究。大業大學食品工程研究所 碩士論文。
廖茂州。2006。太魯閣國家公園海濱植被之研究。國立花蓮教育大學生態與環境教育研究所 碩士論文。
劉業經、呂福原及歐辰雄。1994。臺灣樹木誌。國立中興大學農學院出版委員會。925 頁。
歐辰雄、呂金誠。2002。崩塌地植物復育手冊。行政院農業委員會林務局。183 頁。
蕭鳳岐。1996。蜂膠生物效用的利用。食品資訊 122: 41-45。
Akopyan, Z. M., G. A. Shakaryan, and S. G. Danielyan. 1970. Sensitivity of microorganism to propolis in some districts of the Armenian S. S. R. Biol. Zh. Armeniya. 23: 70-74.
Albrecht, M., H. Frerick, U. Kuhn, and A. Strenge-Hesse. 1992. Therapy of toxic liver disease with Legalon. Zeitschrift. Fuer. Klinis. Med. 47: 87-92.
Amoros, M., C. M. Simoes, L. Girre, F. Sauvager, and M. Cormier. 1992. Synergistic effect of flavones andflavonols against herpes simplex virus type I in cell culture. Comparison with the antiviral activity of propolis. J. Nat. Prod. 55: 1732-1740.
Antúnez, K., J. Harriet, L. Gende, M. Maggi, M. Eguaras, and P. Zunino. 2008. Efficacy of natural propolis extract in the control of American Foulbrood. Vet. Microbiol. 131: 324-331.
Asaka, Y., A. Ohsaki, T. Kubota, Y. Matsukawa, Y. Satomi, and H. Nishino. 1992. 5, 7, 3', 4'-Tetrahydroxy-3-methoxyflavone, a potent anti-tumor promoter isolated from Gnaphalium indicum. J. Kyoto Prefectural Univ. Med. 101: 353-359.
Ayres, D. C., M. C. Marcucci, and S. Giorgio. 2007. Effects of Brazilian propolis on Leishmania amazonensis. Mem. Inst. Oswaldo. Cruz. 102: 215-220.
Babenko, N. A., and E. G. Shakhova. 2008. Effects of flavonoids on sphingolipid turnover in the toxin-damaged liver and liver cells. Lipids Health Dis. 7:1.
Bankova, V. 2005a. Recent trends and important developments in propolis research. Evid. Based Compl. Altern. Med. 2: 29-32.
Bankova, V. 2005b. Chemical diversity of propolis and the problem of standardization. J. Ethnopharmacol. 100: 114-117.
Bankova, V. and M. C. Marcucci. 2000. Standardization of propolis: present status and perspectives. Bee World 81: 182-188.
Bankova, V., S. L. De Castro, and M. C. Marcucci. 2000. Propolis: recent advances in
chemistry and plant origin. Apidologie 31: 3-15.
Bankova, R., R. Christov, G. Stoev, and S. Popov. 1992. Determination of phenolics from propolis by capillary gas chromatography. J. Chromat. 607: 150-153.
Banskota, A. H., Y. Tezuka, and S. Kadota. 2001. Recent progress in pharmacological research of propolis. Phytother. Res. 15: 561-571.
Basnet, P., M. Matsuno, and R. Neidlein. 1997. Potent free radical scavenging activity of propol isolated from Brazilian propolis. Z. Naturforsch. 52: 828-833.
Basnet, P., K. Matsushige, K. Hase, S. Kadota, and T. Namba. 1996. Four di-O-caffeoyl quinic acid derivatives from propolis. Potent hepatoprotective activity in experimental liver injury models. Biol. Pharm. Bull. 19: 1479-1484.
Bastos, E. M., M. Simone, D. M. Jorge, A. E. Soares, and M. Spivak. 2008. In vitro study of the antimicrobial activity of Brazilian propolis against Paenibacillus larvae. J. Invert. Pathol. 97: 273-281.
Boyanova, L., R. Kolarov, G. Gergova, and I. Mitov. 2006. In vitro activity of Bulgarian propolis against 94 clinical isolates of anaerobic bacteria. J. Anaerobe. 12: 173-177.
Brinkworth, R. I., M. J. Stoermer, and D. P. Fairlie. 1992. Flavones are inhibitors of HIV-1 proteinase. Biochem. Biophysic. Res. Com. 188: 631-637.
Buckshee, K., D. Takkar, and N. Aggarwal. 1997. Micronized flavonoid therapy in internal hemorrhoids of pregnancy. Int. J. Gynaecol. Obstet. 57: 145-151.
Burdock, G. A. 1998. Review of the biological properties and toxicity of bee propolis. Food Chem. Toxicol. 36: 347-363.
Chang, C. C., M. H. Yeh, H. M. Wen, and J. C. Chern. 2002. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J. Food Drug Anal. 10: 178-182.
Chen, J. H., Y. Shao, M. T. Huang, C. K. Chin, and C. T. Ho. 1996. Inhibitory effect of caffeic acid phenethyl ester on human leukemia HL-60 cells. Cancer Lett. 108: 211-214.
Chen, C. N., C. L. Wu, and J. K. Lin. 2007. Apoptosis of human melanoma cells induced by the novel compounds propolin A and propolin B from Taiwanese propolis. Cancer Lett. 245: 218-31.

Chen, C. N., C. L. Wu, and J. K. Lin. 2004a. Propolin C from propolis induces apoptosis through activating caspases, Bid and cytochrome c release in human melanoma cells. Biochem. Pharmacol. 67: 53-66.
Chen, C. N., M. S. Weng, C. L. Wu, and J. K. Lin. 2004b. Comparison of radical scavenging activity, cytotoxic effects and apoptosis induction in human melanoma cells by Taiwanese propolis from different sources. Evid. Based Compl. Altern. Med. 1: 175-185.
Chen, C. N., C. L. Wu, H. S. Shy, and J. K. Lin. 2003. Cytotoxic prenylflavanones from Taiwanese propolis. J. Nat. Prod. 66: 503-506.
Chen, Y. W., and K. K. Ho. 2001. The composition and biological activities of propolis from Taiwan. Sixth International Conference on Apitherapy Healthcare, Daegu, Korea. pp.396-410.
Chen, Y. W., S. W. Wu, K. K. Ho, S. B. Lin, C. Y. Huang, and C. N. Chen. 2008. Characterization of Taiwanese propolis collected from different seasons and locations. J. Sci. Food Agric. 88: 412-419.
Cholbi, M. R., M. Paya, and M. J. Alcaraz. 1991. Inhibitory effects of phenolic compounds on carbon tetrachloride induced microsomal lipid peroxidation. Exper. 47: 195-199.
Cizmarik, J. and J. Trupl. 1976. Effect of propolis on bacteria. Phar. 31: 656-657.
Cunha, I. B. S., A. C. H. F. Sawaya, F. M. Caetano, M. T. Shimizu, M. C. Marcucci, F. T. Drezza, G. S. Povia, and P. O. Carvalho. 2004. Factors that influence the yield and composition of Brazilian propolis extracts. J. Braz. Chem. Soc. 15: 964-970.
Daugsch, A., C. S. Moraes, P. Fort, and Y. K. Park. 2008. Brazilian red propolis-chemical composition and botanical origin. Evid. Based Compl. Altern. Med. 5: 435-441.
Dorta, D. J., A. A. Pigoso, F. E. Mingatto , T. Rodrigues, C. R. Pestana, S. A. Uyemura, A. C. Santos, and C. Curti. 2008. Antioxidant activity of flavonoids in isolated mitochondria. Phytother Res. 22: 1213-1218.
Dumitrescu, M., V. Esanu, and I. Crisan. 1992. Mechanisms of the anti-herpetic activity of aqueous extract of propolis. I. Antioxidant activity in human fibroblast cultures. Rom. J. Virol. 43: 165-173.
Dumitrescu, M., I. Crisan, and V. Esanu. 1993. Mechanism of the anti-herpetic activity of aqueous extract of propolis. II , Activity of lectins. from the aqueous extract of propolis. Rom. J. Virol. 44: 49-54.
Duran, G., N. Duran, G. Culha, B. Ozcan, H. Oztas, and B. Ozer. 2008. In vitro antileishmanial activity of Adana propolis samples on Leishmania tropica: a preliminary study. Parasitol. Res. 102: 1217-25.
Freitas, S. F., L. Shinohara, J. M. Sforcin, and S. Guimarães. 2006. In vitro effects of propolis on Giardia duodenalis trophozoites. Phytom. 13: 170-5.
Friedman, M. 2007. Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas. Mol. Nutr. Food Res. 51: 116-134.
Fuliang, H. U., H. R. Hepburn, H. Xuan, M. Chen, S. Daya, and S. E. Radloff. 2005. Effects of propolis on blood glucose, blood lipid and free radicals in rats with Diabetes mellitus. Pharmacol. Res. 51: 147-52.
Gabrys, J., Z. Konecki, W. Krol, S. Scheller, and J. Shani. 1986. Free amino acids in bee live product (propolis) as identified and quantified by gas-liquid chromatography. Pharmacol. Res. 18: 513-518.
Ghisalberti, E.L. 1979. Propolis: A review. Bee World 60: 59-84.
González, R., D. Remirez, S. Rodriguez, A. González, O. Ancheta, N. Merino, and C. Pascual. 1994. Hepatoprotective effects of propolis extract on paracetamol-induced liver damage in mice. Phytother. Res. 8: 229-232.
González, R., I. Corcho, D. Remirez, S. Rodriguez, O. Ancheta, N. Merino, A. González, and C. Pascual. 1995. Hepatoprotective effects of propolis extract on carbon tetrachloride-induced liver injury in rats. Phytother. Res. 9: 14-17.
Greenaway, W., T. Scaysbrook, and F. R. Whately, 1990. The composition and plant origin of propolis: a report of work at Oxford. Bee World 71:107-118.
Grecianu, A., and V. Enciu. 1976. Activity in vitro of propolis against bacterial strains of animal origin. Institutal. Agronomic. Ion. Ionescu. Bradé (Zootehnie. Medicina Veterinara). 90-92.
Grunberger, D., R. Banerjee, K. Eisinger, E. M. Oltz, L. Efros, M. Caldwell, V. Estevez, and K. Nakanishi. 1988. Preferential cytotoxicity on tumor cells by caffeic phenethyl ester isolated from propolis. Exper. 44: 230-232.
Gyorgy, I., S. Antus, A. Blazovics, and G. Foldiak. 1992. Substituent effects in the free radical reactions of silybin: Radiation-induced oxidation of the flavonoid at neutral pH. Intern. J. Radiat. Bio. 61: 603-609.
Harish, Z., Rubinstein, A., Golodner, M., Elmaliah, M. and Mizrachi, Y. 1997. Suppression of HIV-1 replication by propolis and its immunoregulatory effect. Drugs Exp. Clin. Res. 23:89-96.
Hatusima, S. 1975. Flora of Ryukyus, Added and Corrected. The Biological Society of Okinawa, Japan pp.373.
Hayashi, K., S. Komura, N. Isaji, N. Ohishi, and K. Yagi. 1999. Isolation of antioxidative compounds from Brazilian propolis: 3, 4-dihydroxy-5- prenylcinnamic acid, a novel potent antioxidant. Chem. Pharm. Bull. 47: 1521–1524.
Hegazi, A. G., F. EI-Berdiny, S. EI-Assily, E. Khashaball, N. Hassan, and S. Popov 1994. Studies on some aspects of antiviral activity. 1. Influence of propolis on Newcastle disease virus. J. Qatar University Science. 14: 18-20.
Hemández, N. M. R., and K. C. Bemal. 1990. Efecto antibiótico del propóleo frente a cepas de Staphylococcus aureus origen clinico humano. Rev. Cu. Farm. 24: 45-50.
Hertog, M. G. L., P. C. H. Hollman, and M. B. Katan. 1992. Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly in the Netherlands. J. Agric. Food Chem. 40: 2379-2383.
Higashi, K. O. and S. L. De Castro. 1995. Effect of different formulations of propolis on mice infected with Trypanosoma cruzi. J. Ethnopharmacol. 46: 55-8.
Huang, N. T. and T. Ferraro. 1992. Phenolic compounds in food and cancer prevention, in phenolic compounds in food and their effect on health II: antioxidants and cancer prevention. Amer. Chem. Soc. 507: 8-34.
Huang, W. J., C. H. Huang, C. L. Wu, J. K. Lin, Y. W. Chen, C. L. Lin, S. E. Chuang, C. Y. Huang, and C. N. Chen. 2007. Propolin G, a prenylflavanone, isolated from Taiwanese propolis, induces caspase-dependent apoptosis in brain cancer cells. J. Agric. Food Chem. 55: 7366-76.
Hui, W. H., M. M. Li, and K. K. Ng. 1975. Euphorbiaceae of Hong Kong. X. Terpenoids and steroids from Macaranga tanarius. Phytochem. 14: 816-817.
Hui, W. H., K. K. Ng, N. Fukamiya, M. Koreeda, and K. Nakanishi. 1971. Isolation and structure of macarangonol, a diterpene ketol from Macaranga tanarius Phytochem. 10: 1617-1620.
Ilhan, A., U. Koltuksuz, S. Ozen, E. Uz, H. Ciralik, and O. Akyol. 1999. The effects of caffeic acid phenethyl ester (CAPE) on spinal cord ischemia/reperfusion injury in rabbits. Eur. J. Cardiothorac. Surg. 16: 458-63.
Ioirich, N., A. Derevici, and A. Petrescu. 1965. Virulicidal action in vitro of alcoholicextracts of drone larvae and propolis. lucr. stiint stat. Cent. Seri. Apic. 5: 107-110.
Irmak, M. K., E. Fadillioglu, S. Sogut, H. Erdogan, M. Gulec, M. Ozer, M. Yagmurca, and M. E. Gozukara. 2003. Effects of caffeic acid phenethyl ester and alpha-tocopherol on reperfusion injury in rat brain. Cell Biochem. Funct. 21: 283-9.
Kamei, H., T. Koide, T. Kojimam, M. Hasegawa, K. Terabe, T. Umeda, and Y. Hashimoto. 1996. Flavonoid mediated tumor growth suppression demonstrated by in vivo study. Cancer Biother. Radiopharm. 11: 193-196.
Kedzia, B., J. Iwaszkiewicz, and B. Geppert. 1988. Pharmacological investigations on ethanolic extract of propolis. Herba. Pol. 34: 243-53.
Khayyal, M. T., M. A. El-Ghazaly, and A. S. El-Khatib. 1993. Mechanisms involved in the antiinflammatory effect of propolis extract. Drugs under Exp. Clin. Res. 19: 197-203.
Koo, H., B. P. F. A. Gomes, P. L. Rosalen, G. M. B. Ambrosano, Y. K. Park, and J. A. Cury. 2000. In vitro antimicrobial activity of propolis and Arnica montana against oral pathogens. Arch. Oral. Biol. 45: 141-8.
Krell, R. 1996. Value-added products from beekeeping. FAO Agricultual services bulletin No. 124 Food and Agriculture Organization of the United Nations Rome.
Krol, W., S. Scheller, Z. Czuba, T. Matsuno, G. Zydowicz, J. Shani, and M. Mos. 1996. Inhibition of neutrophils' chemiluminescence by ethanol extract of propolis (EEP) and its phenolic components. J. Ethnopharmacol. 55: 19-25.
Kujumgiev, A., I. Tsvetkova, Y. Serkedjieva, V. Bankova, R. Christov, and S. Popov. 1999. Antibacterial, antifungal and antiviral activity of propolis of different geographic origin. J. Ethnopharmacol. 64: 235-40.
Kumazawa, S., J. Nakamura, M. Murase, M. Miyagawa, M. R. Ahn, and S. Fukumoto. 2008. Plant origin of Okinawan propolis: honeybee behavior observation and phytochemical analysis. Naturwiss. 95: 781-6.
Kumazawa, S., R. Ueda, T. Hamasaka, S. Fukumoto, T. Fujimoto, and T. A. Nakayama. 2007. Antioxidant prenylated flavonoids from propolis collected in Okinawa, Japan. J. Agric. Food Chem. 55: 7722-7725.
Kumazawa, S., H. Goto, T. Hamasaka, S. Fukumoto, T. Fujimoto, and T. A. Nakayama. 2004. A new prenylated flavonoid from propolis collected in Okinawa, Japan. Biosci. Biotechnol. Biochem. 68: 260-262.
Kumazawa, S., M. Yoneda, I. Shibata, J. Kanaeda, T. Hamasaka, and T. Nakayama. 2003. Direct evidence for the plant origin of Brazilian propolis by the observation of honeybee behavior and phytochemical analysis. Chem. Pharm. Bull. 51: 740-2.
Lee, Y. J., P. H. Liao, W. K. Chen, and C. Y. Yang. 2000. Preferential cytotoxicity of caffeic acid phenethyl ester analogues on oral cancer cells. Cancer Lett. 153: 51-56.
Lim, T. Y., Y. Y. Lim, and C. M. Yule. 2008. Evaluation of antioxidant, antibacterial and anti-tyrosinase activities of four Macaranga species. J. Food Chem.
Lin, J. H., G. Nonaka, and I. Nishioka. 1990. Tannins and related compounds. XCIV. Isolation and characterization of seven new hydrolyzable tannins from the leaves of Macaranga tanarius (L.) Muell. et Arg. Chem. Pharm. Bull. 38: 1218-1223
Lin, S. C., C. Y. Chung, C. L. Chiang, and S. H. Hsu. 1999. The influence of propolis ethanol extract on liver microsomal enzymes and glutathione after chronic alcohol administration. Am. J. Chin. Med. 27: 83-93.
Lu, H. Q., B. Niggemann, and K. S. Zanker. 1996. Suppression of the proliferation and migration of oncogenic ras-dependent cell lines, cultured in a three-dimensional collagen matrix, by flavonoid-structured molecules. J. Cancer Res. Clin.Oncol. 122: 335-342.
Lu, L. C., Y. W. Chen, and C. C. Chou. 2003. Antibacterial and DPPH free radical-scavenging activities of ethanol extract of propolis collected in Taiwan. J. Food Drug Anal. 11: 277-282.
Lu, L. C., Y. W. Chen, and C. C. Chou. 2005. Antibacterial activity of propolis against Staphylococcus aureus. Int. J. Food Microbio. 102: 213-220.
Manolova, N., V. Maximova, Z. Manolova, I. Stoilova, E. Korczak, and A. Denis. 1987. Immunobiological effect of propolis. I. Effect on Cellular immunity. Acta. Microbiol. Bulg. 21: 76-81.
Marcucci, M. C. 1995. Propolis: chemical composition, biological properties and therapeutic activity. Apidol. 26: 83-99.
Marcucci, M.C., and V. S. Bankova 1999. Chemical composition, plant origin and biological activity of Brazilian propolis. Current Topics in Phytochem. 2: 115-123.
Marquez, N., R. Sancho, A. Macho, M. A. Calzado, B. L. Fiebich, and E. Munoz. 2004. Caffeic acid phenethyl ester inhibits T-cell activation by targeting both nuclear factor of activated T-cells and NF-κB transcription factors. J. Pharmacol. Exp. Ther. (JPET). 308: 993-1001.
Matsunami, K., I. Takamori, T. Shinzato, M. Aramoto, K. Kondo, H. Otsuka, and Y. Takeda. 2006. Radical-scavenging activities of new megastigmane glucosides from Macaranga tanarius (L.) MULL.-ARG. Chem. Pharm. Bull (Tokyo). 54: 1403-1407.
Matsuno, T. 1995. A new clerodane diterpenoid isolated from propolis. Z. Naturf. 50: 93-97.
Matsuno, T., and O. Efeito. 1997. Terapêutico da Própolis. Impressão Abaeté: São Paulo.
Matsushige, K., I. T. Kusumoto, Y. Yamamoto, S. Kadosa, and T. Namba. 1995. Quality evaluation of propolis. 1. A comparative study on radical scavenging effects of propolis and vespae nidus. J. Trad. Med. 12: 45-53.
Matsushige, K., P. Basnet, S. Kadosa, and T. Namba. 1996. Potent free radical scavenging activity of dicaffeoyl quinic acid derivatives from propolis. J. Trad. Med. 13: 217-228.
Menezes, H., J. M. Alvarez, and E. Almeida. 1999. Mouse ear edema modulation by different propolis ethanol extracts. Arzneimittelf. 49: 705-707.
Merino, N., R. González, A. González, and D. Remirez. 1996. Histopathological evaluation on the effect of red propolis on liver damage induced by CCl4 in rats. Arch. Med. Res. 27: 285-289.
Mirzoeva, O. K., and P. C. Calder. 1996. The effect of propolis and its components on eicosanoid production during the inflammatory response, Prostaglandins Leukot. Essent. Fat. Acids 55: 441-449.
Mitamura, T., T. Matsuno, S. Sakamoto, M. Maemure, H. Kudo, S. Suzuki, K. Kuwa, S. Yoshimura, S. Sassa, T. Nakayama, and H. Nagasawa. 1996. Effect of a new clerodane diterpenoid isolated from propolis on chemically induced skin tumors in mice. Antica. Res. 16: 2669-2672.
Miyataka, H., M. Nishiki, H. Matsumoto, T. Fujimoto, M. Matsuka and T. Satoh, 1997. Evaluation of propolis. 1. Evaluation of Brazilian and Chinese propolis by enzymatic and physico-chemical methods. Biol. Pharm. Bull. 20: 496-501.
Moreira, T. F. 1986. Chemical composition of propolis: vitamins and amino acids. Rev. Bras. Farmacogn. 1: 12-9.
Moriyasu, J., S. Arai, R. Motoda, and M. Kurimoto. 1994. In vitro activation of mouse macrophage by propolis extract powder. Bioth. 8: 364-5.
Nakajima, Y., M. Shimazawa, S. Mishima, and H. Hara. 2007. Water extract of propolis and its main constituents, caffeoylquinic acid derivatives, exert neuroprotective effects via antioxidants actions. Life Sci. 80: 370-377.
Nagai, T., R. Inoue, H. Inoue, and N. Suzuki. 2002. Preparation and antioxidant properties of water extract of propolis. J. Food Chem. 80: 29-33.
Nagy, E., V. Papay, G. Litkei, and Z. Dinya. 1986. Investigation of the chemical constituents, particularly the flavonoid components, of propolis and Populi gemma by the GC/MS method. Studies in Organic Chemistry 23: 223-232.
Omori, H., Y. Nio, Y. Minari, H. Takeda, Y. Sato, M. M. Song, N. Hirahara, S. Sumi, and K. Tamura. 1996. Anti-tumor effects of quinolinone derivatives, vesnarinone (OPC-8212) on human pancreas cancer cell lines. J. Japan Society for Cancer Therapy 31: 446-455.
Orsolić, N., S. Terzić, Z. Mihaljević, L. Sver, and I. Basić. 2005. Effects of local administration of propolis and its polyphenolic compounds on tumor formation and growth. Biol. Pharm. Bull. 28: 1928-1933.
Ota, C., C. Unterkircher, V. Fantinato, and M. T. Shimizu. 2001. Antifungal activity of propolis on different species of Candida. Mycoses. 44: 375-8.
Ozer, M. K., H. Parlakpinar, and A. Acet. 2004. Reduction of ischemia-reperfusion induced myocardial infarct size in rats by caffeic acid phenethyl ester (CAPE). Clin. Biochem. 37: 702–705.
Ozturk, F., E. Kurt, M. Cerci, L. Emiroglu, U. Inan, M Turker. and S. Ilker. 2000. The effect of propolis extract in experimental chemical corneal injury. Oph. Res. 32: 13-18.
Paintz, M., and Y. Metzner, 1979. Zur lokalanasthetischen wirkung von propolis und einigen inhaltsstoffen. Pharmazi 34: 839-841.
Par, A. 1992. Pathogenesis and management of alcoholic liver injury. Acta. Physiol. Hung. 80: 325-350.
Park, E. H. and J. H. Kahng. 1999. Suppressive effects of propolis in rat adjuvant arthritis. Arch. Pharm. Res. 22: 554-558.
Park, Y. K., J. F. Paredes-Guzman, C. L. Aguiar, S. M. Alencar, and F. Y. Fujiwara. 2004. Chemical constituents in Baccharis dracunculifolia as the main botanical origin of southeastern Brazilian propolis. J. Agric. Food Chem. 52: 1100-1103.
Park, Y. K., and M. Ikegaki. 1998. Preparation of Water and Ethanolic Extracts of Propolis and Evaluation of the Preparations. Biosi. Biotech. Biochem. 62: 2230-2232.
Pecking, A. P. 1995. Evaluation of lymphoscintigraphy of the effect of a micronized
flavonoid fraction (Daflon 500 mg) in the treatment of upper limb lymphedema. Int. Angiology 14: 39-43.
Pepeljnjak, S., D. Maysinger, and I. Jalsenjak. 1982. Effect of propolis extract on some fungi. Pharmazie 50:165-167.
Phommart, S., P. Sutthivaiyakit, N. Chimnoi, S. Ruchirawat, and S. Sutthivaiyakit. 2005. Constituents of the leaves of Macaranga tanarius. J. Nat. Prod. 68: 927-930.
Pietta, P. G., C. Gordana, and A. M. Pietta. 2002. Analytical methods for quality control of propolis. Fitoterapia 73: 7-20.
Rhajaoui, M., H. Oumzil, M. Faid, M. Lyagoubi, M. Elyachioui, and A. Benjouad. 2001. Antibacterial activity of a moroccan propolis extracts. Sci. Letters. 3: 9-16.
Sahinler, N., and O. Kaftanoglu. 2005. Natural product propolis: chemical composition. Nat. Prod. Res. 19: 183-8.
Salatino, A., E. W. Teixeira, G. Negri, and D. Message. 2005. Origin and chemical variation of Brazilian propolis. Evid Based Compl. Altern. Med. 2: 33-8.
Scazzocchio, F., F. D. D'Auria, D. Alessandrini, and F. Pantanella. 2006. Multifactorial aspects of antimicrobial activity of propolis. Microbiol. Res. 161: 327-33.
Schmidt, J. O. 1996. Bee products: chemical composition and application. Bee products 2: 15-26.
Serkedjieva, J., N. Manolova, and V. Bankova. 1992. Anti-influenza virus effect of some propolis constituents and their analogues (esters of substituted cinnamic acids). J. Nat. Prod. 55: 294-297.
Sforcin, J. M., A. Jr. Fernandes, C. A. Lopes, V. Bankova, and S. R. Funari. 2000. Seasonal effect on Brazilian propolis antibacterial activity. J. Ethnopharmacol. 73: 243-249.
Sharma, M., K. K. Pillai, S. Z. Husain, and D. K. Giri. 1997. Protective role of propolis against alcohol-carbon tetrachloride-induced hepatotoxicity in rats. Indian J. Pharmacol. 29: 76-81.
Sidwell, R. W., J. H. Huffman, B. J. Moscon, and R. P. Warren. 1994. Influenza virus-inhibitory effects of intraperitoneally and aerosol-administered SP-303, a plant flavonoid. Chemoth. 40: 42-50
Silici, S., Koç, N. A., Ayangil, D. and Çankaya, S. 2005. Antifungal activities of propolis collected by different races of honeybees against yeasts isolated from patients with superficial mycoses. J. Pharmacol. Sci. 99: 39-44.
Silva, B. B., P. L. Rosalen, J. A. Cury, M. Ikegaki, V. C. Souza, A. Esteves, and S. M. Alencar. 2008. Brazilian red propolis-chemical composition and botanical origin. Evid Based Complement Alternat Med. 5: 313-316.
Sonmez, S., L. Kirilmaz, M. Yucesoy, B. Yücel, and B. Yilmaz. 2005. The effect of bee propolis on oral pathogens and human gingival fibroblasts. J. Ethnopharmacol. 102: 371-6.
Starzyk, J., S. Scheller, J. Szaflarski, M. Moskwa, and A. Stojko. 1977. Biological properties and clinical application of propolis. II. Studies on the antiprotozoan activity of ethanol extract of propolis. Arzneimittelf. 27: 1198-1199.
Takeda, Y. 2006. Radical-scavenging activities of new megastigmane glucosides from Macaranga tanarius (L.) MULL.-ARG. Chem. Pharm. Bull. 54: 1403-1407.
Tatefuji, T., H. Yamauchi, M. Ikeda, S. Ando, and M. Kurimoto. 1993. Effect of propolis obtained in Brazil on infectivity of viruses. Shoyakugaku. Zasshi. 47: 60-64.
Teixeira, E. W., G. Negri, R. M. Meira, D. Message, and A. Salatino. 2005. Plant origin of green propolis: bee behavior, plant anatomy and chemistry. Evid. Based. Compl. Altern. Med. 2: 85-92.
Tikhonov, A. I., and I. N. S. Mamontova. 1987. Production and study of a lyophilized phenolic polysaccharide preparation from propolis. Farmatsev. Zhurnal. 3: 67-68.
Topalkara, A., A. Vural, Z. Polat, M. I. Toker, M. K. Arici, F. Ozan, and A. Cetin. 2007. In vitro amoebicidal activity of propolis on Acanthamoeba castellanii. J. Ocul. Pharmacol. Ther. 23: 40-5.
Tosi, B., A. Donini, C. Romagnoli, and A. Bruni. 1996. Antimicrobial activity of some commercial extracts of propolis prepared with different solvents. Phytother. Res. 10: 335-336.
Tseng, M. H., C. H. Chou, Y. M. Chen, and Y. H. Kuo. 2001. Allelopathic prenylflavanones from the fallen leaves of Macaranga tanarius. J. Nat. Prod. 64: 827–828.
Vereckei, A., E. Feher, I. Gyorgy, G. Foldiak, M. Toth, H. Toncsev, and J. Feher. 1991. The possibilities of antioxidant therapy in the prevention of amiodarone side effects. Orv. Hetil. 132: 1265-1268.
Vokhonina, T. V., L. G. Breeva, R. N. Bodrova, and E. S. Dushkova. 1969. Some physical and chemical antimicrobial characteristics of propolis and extracts. 22nd Int. Beekeep Congr. Summ. 211-217.
Weng, M. S., C. H. Liao, C. N. Chen, C. L. Wu, and J. K. Lin. 2007. Propolin H from Taiwanese propolis induces G1 arrest in human lung carcinoma cells. J. Agric. Food Chem. 55: 5289-5298.
Woisky, R. G., and A. Salatino. 1998. Analysis of propolis: some parameters and procedures for chemical quality control. J. Apicul. Res. 37: 99-105.
Yang, H. Y., C. M. Chang, Y. W. Chen, and C. C. Chou. 2006. Inhibitory effect of propolis extract on the growth of Listeria monocytogenes and the mutagenicity of 4-nitroquinoline-N-oxide. J. Sci. Food Agric. 86: 937-943.