高多酚草本植物萃取物對人類週邊淋巴球細胞的免疫調節作用

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Title:	高多酚草本植物萃取物對人類週邊淋巴球細胞的免疫調節作用 Immunomodulating effect of herbal extracts with higher polyphenolic contents in human peripheral blood lymphocyte
Authors:	徐碩妤
Contributors:	陳師瑩戴三堡嘉南藥理科技大學：營養與保健科技研究所
Keywords:	海巴戟天葉香蓼免疫調節作用人類週邊血液淋巴球虎杖根 Morinda citrifolia Polygnum odoratum Polygonum cuspidatum root immunomodulation human blood lymphocytes
Date:	2010
Issue Date:	2010-12-30 09:48:22 (UTC+8)
Abstract:	失衡的免疫調節作用與過度的發炎反應，在人類許多疾病的發生上扮演一個十分重要的角色，例如常見的癌症、心血管疾病、老化等等。天然植物大抵含有酚類物質，過去文獻提及在 in vitro 或 in vivo 試驗中主要可能藉由調控細胞核轉錄因子 NF-κB 活性，與影響細胞激素間的平衡關係，而具有抗發炎活性。過去研究發現，乙醇或水萃取而來的海巴戟天葉 (Morinda citrifolia leaf, LES)、香蓼 (Polygnum odoratum, Po) 或虎杖根部 (Polygonum cuspidatum root, PcE) 萃取物，除了富含高多酚類化合物之外，更擁有良好抗氧化活性，因此本研究試著更進一步的純化這三種天然物質，並探究其活性成分是否也同時具有抗發炎之免疫調節特性。嘉南藥理科技大學保健中心代為隨機招募約30人次，20-40歲的健康成人血液；隨後將採集而來的血液分離出人類週邊血液淋巴球細胞 (PBMC) 進行初代培養以做為實驗受試細胞之用。本研究試圖假設上述三種天然萃取物對 PBMC 可能具免疫調節效果，因此先給予天然萃取物後，再施予誘導劑 LPS 以啟動發炎模式的產生，接著評估是否對發炎媒介物 TNF - α、IL-6、IL-1β 與 IL-10 具調節作用，甚至深入探討與細胞核中活化型 NF- κB 蛋白之間的關係。主要實驗結果發現 : (1) LES 在濃度 200-300 μg/mL 時能明顯抑制 IL-6、IL-1α 與 NF-κB 的表現，而具有免疫調節作用。(2) 自 LES 中純化而來的 Fraction I 具良好的抗發炎活性，在 10 ng/mL-1 μg/mL 時可以向下調節 TNF-α，以及在1-10 μg/ml時可以向下調節 IL-6，不過在 10 μg/ml 時卻對抗發炎細胞激素 IL-10具有向上調節之效果。(3) PoW 或 PoM 能抑制 TNF-α 及細胞核中 NF-κB 蛋白的表現，不過在濃度 12.5 μg/mL 以上時具明顯細胞毒性。(4) PcE 在低濃度 100 ng/mL-10 μg/mL 時幾乎不具有細胞激素調節效果，甚至當濃度在 3.1 μg/mL 以上時，就有明顯的細胞毒性。無論如何，強而有力的潛在性抗發炎因子及酚類化合物與分子機制間的調控關係仍有待進一步的探討確認，以應用於預防及治療之上。 Excessive inflammation is considered as a critical factor in many human diseases, including cancer, cardiovascular diseases, and aging. Compounds derived from botanic sources, such as phenolic compounds, have shown anti-inflammatory activity in vitro and in vivo. Recent data suggest that polyphenols can express anti-inflammatory activity by modulation of pro-inflammatory gene expression, mainly by acting through NF-κB or cytokines. Based on our researches, ethanolic or aqueous extracts from Morinda citrifolia leaf (LES), Polygnum odoratum (Po), and Polygonum cuspidatum root (Pc) all exhibit higher polyphenolic contents and reveal that these compounds play a part in antioxidative activities. Hence, the three herbs were worth further research in purification and identification of active compounds, and deeply investigation for anti-inflammatory effects. About 30 healthy volunteers 20–40 y of age will be recruited randomly through Health Care Center in Chia-Nan University of Pharmacy and Science. Herbal extracts were used to study the inhibition of the proinflammatory cytokines including TNF-α, IL-6, IL-1β and IL-10 and the expressions of NF-κB in human peripheral blood lymphocyte (PBMC) stimulated by LPS, respectively. Our main findings were: (1) LES at 200-300 μg/mL had immunomodulatory effects on cytokine release from activated PBMC with a marked reduction in IL-6, IL-1β and NF-κB expression. (2) Fraction I purified from LES had immunomodulatory effects on cytokine release from activated PBMCs with down-regulation of TNF- α at 10 ng/mL-1 μg/mL and IL-6 at 1-10 μg/mL, but up-regulation of IL-10 at 10 μg/mL. Fraction I showed potent anti-inflammatory effects of activated PBMCs. (3) PoM or PoW could decrease the cytokine stimulating effects of TNF- α and NF-κB expression. However, they above 12.5 μg/mL had significantly cytotoxicity. (4) PcE at 100 ng/mL-10 μg/mL had virtually no effects on cytokines, and above 3.1 μg/mL had significantly cytotoxicity. However, a clear understanding of the molecular mechanisms of action of phenolic compounds is crucial in the valuation of these potent molecules as potential prophylactic and therapeutic agents.
Relation:	校內校外均不公開，學年度：98，141頁
Appears in Collections:	[Dept. of Health and Nutrition (including master's program)] Dissertations and Theses

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