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標題: 管花肉蓯蓉中松果菊苷具有類飢餓素刺激生長激素分泌之活性分析
Bioactivity of Echinacoside in Cistanche tubulosa, mimicking ghrelin to stimulate growth hormone secretion
作者: 吳婕如
Chieh-Ju Wu
關鍵字: 管花肉蓯蓉
Cistanche tubulosa
phenylethanoid glycosides
primary pituitary cell culture
growth hormone
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摘要: 肉蓯蓉(Cistanche spp. )為中國典籍中珍貴的中藥之一,其補腎壯 陽、延年益壽、潤腸通便等記載,使其享有'沙漠人參'之美譽。近代 藥理學研究更指出其主要成分苯乙醇苷類(phenylethanoid glycosides;PhGs)具保肝、抗氧化、抗衰老、神經保護、免疫調節、增強學習記憶能力等功效,並以含量最高之松果菊苷(Echinacoside;ECH)作為品管標準。生長激素(growth hormone;GH)為人類生長及延緩老化所需之關鍵荷爾蒙,然而其直接施用易引起水腫、關節疼痛等副作用,故近年文獻中重視於開發更上游之荷爾蒙:飢餓素(ghrelin)及類飢餓素藥物(ghrelin-liked)來達到全面性調控生長激素、促進腸胃蠕動、增加食慾及記憶力等療效。本實驗室前人研究中發現類飢餓素成分:茶飢素(Teaghrelin)及銀杏飢素(Ginkgoghrelin),其分子結構與管花肉蓯蓉的 Echinacoside 相類似,根據先前分子對接(molecular docking)模擬後,其 glycosides 及 p-coumaric acid 官能基能與飢餓素接受器結合,達到與飢餓素(ghrelin)相同調控路徑,進而促進生長激素分泌之生理功效。故本實驗使用Sephadex LH-20管柱於管花肉蓯蓉萃取液(CTE)純化出Echinacoside,並透過活體 Sprague Dawley (SD)品系大鼠腦下垂體之初代細胞進行生長激素分泌測詴。以人工合成之飢餓素受器促效劑 (growth hormone-releasing peptide 6;GHRP-6)做為生長激素分泌之正對照組,於初步實驗結果中顯示(10-5-10-8 M )的濃度下 Echinacoside具促進分泌生長激素能力,並呈現藥物濃度依賴性。在 30 分鐘,Echinacoside 在 10-5 M 濃度下相較於正對照組有較高的分泌量。透過GHS-R1a antagonist 證明與 GHRP-6 走相同的分泌路徑。最後由分子對接(molecular docking)模擬,去臆測Echinacoside 與接受器互相鍵結的接位。進一步探討利用 Echinacoside 作為新類飢餓素藥物之可能性。
Cistanche spp. , the ginseng of desert, has been recorded to possess several biological activities, such as constipation relieving, longevity and aphrodisiac property in traditional Chinese pharmacopoeia. Recent pharmacological studies showed that the major active ingredients in Cistanche spp. are phenylethanoid glycosides (PhGs), which possess various effects, e.g., antioxidative, hepatoprotective and neuroprotective activities as well as enhancing learning capacity. Echinacoside (ECH) is the most abundant constituent in PhGs, and constantly regarded as a reference compound for the quality control in CT. Growth hormone is an essential hormone that promotes human growth and anti-aging; however, direct application of growth hormone indicates serious side effects like edema and arthralgia. Therefore, it has been searched for drugs that stimulate growth hormone secretion, such as ghrelin and ghrelin-liked molecules. In previous studies, two natural compounds, teaghrelin and ginkgoghrelin, showed biological activities similar to ghrelin. The chemical structure of ECH consisting two glucosides, one rhamnose, one caffeic acid, and one hydroxytyrosol is comparable to teaghrelin and Ginkgoghrelin. Therefore, I propose that ECH may be able to interact with ghrelin receptors. In this study, I purified ECH natural compound from CT by using Sephadex LH-20 column and then assayed growth hormone secretion by treating pituitary cells which were isolated from male Sprague−Dawley rats. Positive control of growth hormone-releasing hormone-6 (GHRP-6) is a synthetic analog of ghrelin which stimulated growth hormone secretion, and the result showed that the concentration (10-5 M -10-8 M) of ECH induced the secretion of growth hormone and have dose dependenced. At 30 minutes, Echinacoside had a higher secretion level in 10-5 M compared with positive control. To demonstrate, Echinacoside have the same secretory pathway as GHRP-6 by GHS-R1a antagonist. Finally, using molecular docking to simulate were performed to determine the binding sites between the Echinacoside and the receptor. Further explore the possibility of using Echinacoside as a new class of ghrelin drugs in the future.
文章公開時間: 2019-08-09
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