桂花化學成分與活性探討

Studies on the Constituents and Bioactivity of Osmanthus fragrans Flowers

張惠欣(Huei-Shin Chang)

指導教授 ： 石麗仙

國立虎尾科技大學/文理學院/生物科技研究所/碩士(2013年)

https://doi.org/10.6827/NFU.2013.00087

摘要

桂花(Osmanthus fragrans)為木犀科(Oleaceae)木犀屬常綠灌木，主要分布於中國、日本和喜馬拉雅山。其根、根皮、花、果實、葉子與種子皆能做為藥用，而花具有化痰、散瘀功效又能治痰飲咳喘、腸風血痢、疝瘕、牙痛與口臭。本研究以人類結直腸腺癌細胞(WiDr cell)做為體外抗發炎活性試驗模式進行活性評估，發現桂花乙醇粗抽物(OFE)具有DPPH清除、抗胞內氧化壓力與抗發炎活性，因此以生物活性為導向進行桂花抗氧化與抗發炎活性成分之分離並探討其生物活性。
將桂花以乙醇熱迴流萃取，乙醇粗抽物再以水和氯仿與正丁醇進行分配萃取，分別得到氯仿層(OFE-C)、正丁醇層(OFE-B)和水層(OFE-W)。正丁醇層具有DPPH清除能力(IC50為74.52 μg/ml)與抑制前發炎因子IL-8之分泌，因此利用管柱層析和高壓液相層析等方法分離出十三個化合物。經文獻比對各化合物的光譜數據，確認十個化合物之構造為mannitol (1), verbascoside (2), oleoside dimethyl ester (3), phillygenin (4), phillyrin (5), (3E)-4-(3,4-dihydroxyphenyl)-but-3-en-2-one (6), tyrosol (7), methyl 4-hydroxycinammate (8), β-sitosterol (9)和 eutigoside A (10)，其中7與8為首次自桂花中分離得到之化合物，而2為桂花的主成分。
分離得到之化合物分別進行DPPH清除能力、胞內抗氧化壓力與IL-8分泌的抗發炎作用，結果顯示2具有DPPH清除能力 (IC50為8.23 μg/ml)；2與3在10 μg/ml時可降低WiDr cell的胞內氧化壓力；4在10 μg/ml具有抑制IL-8分泌。1-3和7在1 μg/ml和2及8在10 μg/ml下能抑H2O2誘導WiDr細胞之IL-8分泌，而4和10對H2O2誘導WiDr細胞之IL-8分泌具有顯著抑制作用劑量依賴性(0.01~10 μg/ml)。由此結果可知，桂花具有抗發炎之潛力，其作用機制仍然在研究中。

關鍵字

桂花 ； 木犀科 ； 抗發炎 ； 抗氧化壓力 ； 白介素-8(IL-8)

並列摘要

Osmanthus fragrans (Oleaceae family) is belonging to the Osmanthus genus and also known as sweet olive. The roots, root barks, flowers, fruits, leaves and seeds have medicinal benefit and mainly distributed in China, Japan, and the Himalayas. The flowers were used to reduce phlegm and stasis, arrest dysentery with blood in bowel, and also used for the treatment of stomachache, diarrhea, hernia, toothache and bad breath.
This study was by using human colon carcinomacells (WiDr cells) to be the anti-inflammatory activity experimental model. The aims of this study were evaluated DPPH radical scavenging activity, antioxidive stress and inhibition of IL-8 secretion of O. fragrans. We have found that the ethanol extract of O. fragrans flowers (OFE) showed DPPH radical scavenging, anti-oxidative stress and inhibition of IL-8 secretion on WiDr cells. Therefore, OFE was dissolved in water, then partitionated with CHCl3 and n-butanol to obtain OFE-C, OFE-B and OFE-W layers, respectively.
According to the bioactivity assay, the OFE-B layer was subjected on column chromatography and high performance liquid chromatography to afford thirteen compounds. Their structures were elucidated by spectroscopic analysis and compared with the literature data, named as mannitol (1), verbascoside (2), oleoside dimethyl ester (3), phillygenin (4), phillyrin (5), (3E)-4-(3,4-Dihydroxyphenyl)-but-3-en-2-one (6), tyrosol (7), methyl 4-hydroxycinammate (8), β-sitosterol (9) and eutigoside A (10). Among of them, 7 and 8 were isolated from OFE for the first time, and the major component was 2.
The compounds were evaluated DPPH radical scavenging, antioxidive stress and IL-8 secretion activities on WiDr cells, which were treated with H2O2 or non-H2O2. The results showed 2 exhibited DPPH radical scavenging activity (IC50 8.23 μg/ml). 2 and 3 can reduced oxidative stress on WiDr cell at 10 μg/ml. 4 can inhibit the IL-8 secrection on WiDr cell at 10 μg/ml. Moreover, compound 1-3 and 7 at 1 μg/ml, and 2 and 8 at 10 μg/ml can inhibit IL-8 secretion on H2O2-induced WiDr cells. Compound 4 and 10 inhibited IL-8 secretion on H2O2-induced WiDr cells in does-dependent manner (0.01 to 10 μg/ml). Based on the above results, O. fragrans may be a potential herbs for anti-inflammation. The mechanism of this effect still need further investigation.

並列關鍵字

Osmanthus fragrans ； Oleaceae ； anti-inflammatory ； anti-oxidive stress ； Interleukin-8 (IL-8)

參考文獻

施益民和呂鋒洲 (1989). 自由基與各種疾病. 當代醫學 16, 399-407.

Ames, B. N. (1983). Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases. Science 221, 1256-1264.

Apel, K., Hirt, H., 2004. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 55, 373-399.

Baranovsky, A., Schmitt, B., Fowler, D. J., and Schneider, B. (2003). Synthesis of new biosynthetically important diarylheptanoids and their oxa- andfluoro-analogues by three different strategies. Synthetic Communications 33, 1019-1045.

Benkrief, R., Ranarivelo, Y., Skaltsounis, A. L., Tillequin, F., Koch, M., Pusset, J., and Sevenet, T. (1998). Monoterpene alkaloids, iridoids and phenylpropanoid glycosides from Osmanthus austrocaledonic. Phytochemistry 47, 825-832.