201249450 六、發明說明： 【發明所屬之技術領域】 本發明係關於桂花的新用途，特別是桂花萃取物在製 • 備抗憂鬱藥物或保健食品之用途。 【先前技術】 桂花（Ο細⑽认财财），又名月桂、木樨。桂花 香氣淡雅且持久，常作為庭園造景之用，其花部可作為茶 葉或食品的香味添加物，用以增加香氣。 在中藥醫典記载，桂花之花部、杲實及根部皆可入 藥，其中，花部具有散寒破結，化痰止咳之作用，對食慾 不振、痰飲咳喘、腸風血痢及經閉腹痛有一定療效，果實 具有暖胃、平肝及散寒之作用，而根部具有祛風濕及散寒 之作用。 ' 請參照中華民國公告第1341205號「由桂花中萃取抗 氣喘物質的方法」專利案，該專利案揭示桂花萃取物具有 抗氣喘效果，能夠減緩呼吸道及肺部慢性發炎症狀，更能 夠舒缓氣喘症狀’以預防及減少氣喘發作。目前未有任何 研究資料證實桂花具有抗憂鬱之效用。 憂鬱症係-種精神官能症，患者通常會有失眠、食欲 不振、體重減輕、注意力不集中、易怒、情緒低落不快樂二 對生活不感興趣及生活常感無助、無盼望、有輕生的念頭， 甚至有自殺的行為。根據世界衛生組織統計，全世 症發病率約為3.1%，目前憂鬱症係全球排名第四大疾病^ 且預估在2025年，憂鬱症將躍升為全球第二大疾病。 201249450 目前臨床上治療憂鬱症的策略除了服用抗憂鬱藥物 之外，患者應接叉專業人員的心理諮商，較佳係二者同時 進行，以掌控患者病情，並藉由諮商過程中逐量減少憂鬱 症藥物之劑量。 憂鬱症藥物之作用機理係透過提高腦中的血清素 (Serotonin )、正腎上腺素（N〇repinephrine )或多巴胺 (Dopamin)的濃度，使腦部的神經生長素 neurotrophic factor)增加，使腦部神經細胞增生更快且更 強壯，避免腦部神經細胞萎縮，而容易產生憂#現象。 目前憂f症藥物中最廣為處方的係選擇性▲清素回 收抑制劑（SSRI) ’其他尚有三環抗憂鬱藥物、四環抗憂 營藥物、可逆性單胺氧㈣素抑_ (Rima)及選擇性血 =素新腎上腺素回收抑制劑（聰）等，醫師可以考慮患 ^否合财其他身體疾錢其年齡麵擇適#之抗㈣ 樂物’以減低抗憂鬱藥物對患者產生之副作用。 不論是何種類型之抗憂龍物， 副作用，例如π乾、便秘、切 ^者帶來介夕 主徘尿困難、頭暈、頭痛、 心跳加速、腸胃不適、噁心、隹岸、 、 您I ^ . ，、、、愿艮欲降低或性功能障 礙4，这㈣侧料使患者不 將藥物用量降低。抗憂必彡 祕’或 (服藥期間通常為3至6個月），使體内持續服用 作用漠度，才能夠有效抑制憂營症的發:持:=物 :抗憂㈣㈣量降低’抗錄藥物無 ^期^ 果，而無法改善憂蠻症的情形。 ^所預期之效 此外，憂鬱症係尚復發的病 、-、勺有50〜80%的患者 201249450 至少會有—次的復發，通常在首次發作後的二至三年内再 次復發。然而，目前為止並無〆種能夠讓㈣症患者長期 服用且生副相之抗憂義物或保健食品，使憂營 症患者此夠藉以穩定控制病情，降低憂鬱症之復發率。 【發明内容】 f發明之主要目的係提供一種桂花萃取物在製備抗 憂鬱藥物或保健食品之用途，—治療有效劑量之桂花萃取 物係可用於製傷治療或預防憂樣症之藥物或保健食品。 、本發明之次一目的係提供一種桂花萃取物之製備方 法’係用以提供—種桂花萃取物，以應用於製備治療或預 防憂鬱症之藥物或保健食品。 發明之又—目的係提供一種用於預防或治療憂鬱 症之醫藥組合物，該醫藥組合物包含桂花萃取物及醫藥學 上可接受之載劑或賦形劑。 ’、 為達到前述發明目的，本發明所運用之技術内容包含 有： 一—一種桂花萃取物，其係藉由下列步驟之方法而製得·· 、一萃取步驟’將桂花浸泡於—溶劑中共同形成—萃取混合 液’以該㈣將該桂花中之抗氧化物質萃取至該萃取混合 =中’-過濾轉，將料取混合液之㈣剩餘物去除， 得到一萃取液；以及—賴步驟，取該萃取液進行乾燥處 理後，以獲得乾燥後之桂花萃取物。 本發明之桂花萃取物巾，該桂花萃取㈣萃取自金 桂、銀桂、四季桂或丹桂之花部或乾燥花部。 1 201249450 、二種如上所述之桂花萃取物，其在製備預防或治療憂 營症藥物或保健食品之用途，該桂花萃取物係具有提升個 體，抗氧化力及麩胱甘肽含量之表現，該桂花萃取物較佳 係來自金桂、銀桂、四季桂或丹桂之花部。 — 種如上所述之桂花萃取物之製備方法，係包含：一 萃取步驟’將桂花浸泡於—溶劑中共同形成—萃取混合 ，由以該溶劑將該桂花#之抗氧化物料取至該萃取混合 二,丄：過齡驟，將該萃取混合液之固_餘物去除， 二後二二，广及一乾燥步驟’取該萃取液進行乾燥處 後以獲仔乾燥後之桂花萃取物。 桂、桂ί萃取物之製備方法中，該桂花較佳係金 銀桂、四季桂或丹桂之花部或乾燥花部。 本發明桂花萃取物之製備方法中 重量比例可以為1:15至1:10。 不化…“劑之 甲醇本ίΓΓ取物之製備方法中，該溶劑較佳為水、 :。乙一、乙一、丁丙炫、己烧、石_ 複進花萃取物之製備方法中，該萃取步驟可以重 本發明桂花萃取物之製備方 乾燥=明蒸發處理或加 濃縮法方式處菌將該萃取液以減壓 一種用於預防或治療憂攀 ⑷一治療纽量之-選自於合物，其包含： 的才土化萃取物；以及(b) 201249450 一醫藥學上可接受之載劑或賊形劑。 —本發明用於預防或治療憂繫症之醫藥組合物中，該醫 藥、且口物|^佳係以卩服方式投予個體，其中，該醫藥組合 物可以選擇為錠劑、膠囊、粉劑、粒劑或液劑。 /本發明躲或治療憂鬱症之醫藥組合物中 ，較佳 係以每公斤體重之麵投予⑽紅桂花萃取物。 【實施方式】 i為讓本發日月之上述及其他目的、特徵及優點能更明顯 易懂’下文轉本發明之較佳實施例，姐合所附圖式， 作詳細說明如下： 人口本發明係一種桂花萃取物在製備抗憂鬱藥物或保健 “之用途係將療有效量之桂花萃取物用於製 備治療或預防㈣症之藥物或健食品，雜花萃取物係 具有提升腦部與憂鬱相關腦區（如海馬迴區）之抗氧化能 力’並促進腦區血液循環，幫助腦部抵抗自由基或其他氧 化物之侵襲，以達到治療或預防憂鬱症。 該桂花萃取物係取金桂（似var. thunbergiO、mk Q〇smanthusfragrans 胤 lat神⑹、四 奪给 i Osmanthus fragrans νΆτ. semperflorens、氟丹隹 (Osmanthusfragram 猶.aurantiacus)等桂^之花部’特 別係經過乾燥後之花部進行抗氧化物質之萃取，本實施例 係取金桂之乾燥花部進行水萃法。 請參照第1圖所示，本發明萃取桂花之製備方法係包 含一萃取步驟S1、一過濾步驟S2及一乾燥步驟S3。 201249450 該萃取步驟si，係將桂花浸泡於一溶劑中共同形成一 萃取混合液’以該溶劑將該桂花中之抗氧化物質萃取至該 萃取混合液中，其中，該溶劑可以為水、甲醇、乙醇、丙 酮、乙烷、丙烷、丁丙烷、己烷、石油醚或苯等溶劑，桂 花與該溶劑之重量比例較佳為丨：15至丨：10。更詳言之， 本貫施例係以水做為溶劑，將桂花浸入水中15分鐘後，將 該萃取混合液煮沸，進行萃取至少丨小時，得該萃取混合 液，該萃取步驟S1較佳係重複進行id次，以完全萃取 出桂花中的抗氧化物質。本實施例係取9 8公斤之桂花， 以150公升之沸水進行萃取丨小時，再浸置丨小時後，取 出萃取過之桂花可再以1〇〇公升之沸水再進行一次萃取， 總共得到約185公升之萃取混合液。本實施例藉由水做為 萃取溶劑，紐花的抗氧化物f萃取至科取混合液中， 能夠使該萃取混合液中保有較多的抗氧化物質。 該過濾步驟S2係將該萃取混合液之固體剩餘物去 除’得到-萃取液。本實關係將該萃取混合液中之固體 剩餘物去除，以減壓濃縮法將鱗取料祕理至便於進 行滅菌之體積，例如以高溫高__進行滅隨，以便 保存萃取得的抗氧化物質’遂進行該乾燥步驟幻。藉此， 有助於保存料取狀抗氧㈣f，並財 曰 之效率。 w =絲麵S3，取鮮轉進行辆處理後，以獲得 2後之桂花萃取物。更詳言之，該乾鱗縣可選擇以 處理、健錢處理、雜處理或加熱乾燥處理 方式進行賴；本實❹m科錢料式進行乾 201249450 理藉此’便可將桂花之抗氧化物質濃縮並乾燥，再依需 求配製成所需濃度以進行後續試驗。 為證實本發明之桂花萃取物在製備抗憂㈣物或保 健食品之用途，取上述水萃法所得之桂花萃取物，進行下 試驗以朗雜花萃輪對抑锻鬱症之個··（A)桂花 萃取物之抗氧㈣質含量賴、⑼桂花萃取物對於改善類 憂鬱動物模紅4鬱情形、（Q腦部及各顧之抗氧化能力 5式|^及(〇)海馬迴區之總抗氧化力及gsh含量。 (A) 桂花萃取物之抗氧化物質含量測試 本具知例係以總抗氧化能力（〇xygen radical ^sorbance capacity，簡稱〇RAC)及總酚含量作為抗氧化 月匕力之測疋扣標。該桂花萃取物之總抗氧化能力為 18.8±0.8mM trolox equivalents，該桂花萃取物之總酚含量 為65.7±〇.77mg GAE/ml ’證實本發明枝桂花萃取物確實含 有抗氧化物質。 (B) 桂花萃取物對於改善類憂鬱動物模式之憂鬱情形 (B1)建立類憂鬱動物模式（Maternal deprivation animal model) 本實施例係以早期隔離模式（MaternaI deprivation model)提供一種類憂鬱動物模式以進行後續抑制憂鬱症之 試驗。飼養妊娠SD大鼠（自樂斯科生物科技股份有限公 司購得之SPF級Sprageu-Dawley rat，簡稱SD大鼠），將 該妊娠SD大鼠分娩後第一天定為第〇天（pND〇)，其中， 忒姓娠SD大鼠生產數隻雄性大鼠，並於其中隨機挑選 實驗組及對照組之雄性SD大鼠。對照組之SD大鼠（簡稱 201249450 CT氣）係以正常方式與母鼠共同飼養；實驗組之sd大鼠 (簡稱MD鼠）則自PND2起至PND14，每天將MD鼠與 母鼠隔離1小時，固定於每日下午5〜6點將該雄性SD幼 a置於另一空間，使其不受外界環境干擾。 至該MD鼠斷奶後（PND23 )，將每四隻雄性SD幼鼠 分為一籠，飼養於溫度（22土lt：)、光照（曰照時間12小 a寺）皆維持於正常範®之環境，自由傲食飼料及水。直到 PND 28後’即可進行類憂t行為之動物試驗。 (B2)類憂鬱行為之動物試驗 根據Porsolte/d.等人(1977)於《Nature》期刊所發表 的研究指出’藉由老鼠於水巾不動（Imm(趙~ )的情形可 以反應出老鼠的行為喊，即憂咖向，特職以在水中 不動的時間長短可做為判定錢之憂_度，目前的基礎 醫學研究巾，水巾獨行為係作為老鼠的錄重要指標。 依照上梯⑽述之方式建立類㈣鼠以進行試驗 (=2财杨實驗，魏_)之_準備— 溫抑)，測試前-天需使老鼠 熱悉水性，將老鼠置入水槽15八 泡照射30分鐘以供乾老鼠，缓並以60W， 驗，將老鼠置入同樣條件之水槽中5 ::二進了水:不動貫 該5分鐘内的掙扎時間，由計算老鼠於 , gp^r^5lI ,. 对碍間5分鐘扣除其掙扎時 不動時間作為其憂鬱指標。 5月參知、第1表所示，太眘# /, 組）及6組勤氣（分別為第二例係取1組CT鼠（第刖 、刀⑴局弟Bi至恥組201249450 VI. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a novel use of osmanthus, in particular to the use of an osmanthus extract for preparing an antidepressant or health food. [Prior Art] Osmanthus fragrans (Ο ( (10) recognizes wealth), also known as Laurel, Mudu. Sweet-scented osmanthus fragrance is light and long-lasting. It is often used as a garden landscaping. Its flower parts can be used as aroma additives for tea leaves or foods to increase aroma. In the Chinese medicine medicinal code, the flower, sweet and root of sweet-scented osmanthus can be used as medicine. Among them, the flower part has the function of dispelling cold and stagnation, relieving phlegm and relieving cough, and has a loss of appetite, phlegm and cough, and wind and blood. Trans-abdominal pain has a certain effect, the fruit has the effect of warming the stomach, calming the liver and dispelling cold, while the root has the effect of rheumatism and dispelling cold. Please refer to the Patent Case of the Republic of China Announcement No. 1341015, “Method for Extracting Anti-Asthmatic Substances from Sweet-scented Osmanthus”. The patent case reveals that the osmanthus extract has anti-asthmatic effect, can slow down the chronic inflammation of the respiratory tract and lungs, and can relieve asthma symptoms. 'To prevent and reduce asthma attacks. There is currently no research data confirming that osmanthus has anti-depressant effects. Depression-type psychosis, patients usually have insomnia, loss of appetite, weight loss, lack of concentration, irritability, low mood, unhappiness, uninterested in life, often unhelpful, lifeless, hopeless, and suicidal The thoughts, even suicidal behavior. According to the World Health Organization, the incidence of the whole world is about 3.1%. Currently, depression is the fourth largest disease in the world. ^ It is estimated that in 2025, depression will rise to the second largest disease in the world. 201249450 At present, the strategy for treating depression in clinical practice, in addition to taking anti-depressant drugs, patients should be in contact with the psychological counseling of professionals, preferably both at the same time to control the patient's condition, and by the negotiation process Reduce the dose of depression medication. The mechanism of action of depression drugs is to increase the concentration of serotonin (Nero-repinephrine) or dopamine (Dopamin) in the brain to increase the neurotrophic factor of the brain. Cell proliferation is faster and stronger, avoiding brain nerve cell atrophy, and is prone to worry. At present, the most widely prescribed drug in the disease is the selective serotonin recovery inhibitor (SSRI). Others have tricyclic antidepressant drugs, tetracyclic anti-depression drugs, reversible monoamine oxygen (tetra) _ ( Rima) and selective blood = prime new adrenaline recovery inhibitor (Cong), etc., the physician can consider the risk of other health problems, the age of the face of the appropriate anti-(four) music "to reduce anti-depression drugs to patients The side effects. No matter what type of anti-defense dragon, side effects, such as π dry, constipation, cut ^ bring the main diarrhea, dizziness, headache, rapid heartbeat, gastrointestinal discomfort, nausea, sputum, you I ^ . , , , , wish to reduce or sexual dysfunction 4, this (four) side material so that patients do not reduce the amount of drugs. Anti-worry must be secretive' or (usually 3 to 6 months during the medication), so that the body can continue to take action indifferent, can effectively suppress the development of the stagnation of the camp: hold: = things: anti-worry (four) (four) reduce the amount of resistance There is no cure for the drug, and it is not possible to improve the situation of sorrow. ^The expected effect In addition, depression is still a recurring disease, -, 50 to 80% of patients with spoons 201249450 at least - a recurrence, usually recurrence within two to three years after the first episode. However, there are no anti-anxiety or health foods that can cause long-term use of the patients with (4) symptoms, so that patients with sorrow can be used to stabilize the disease and reduce the recurrence rate of depression. SUMMARY OF THE INVENTION The main purpose of the invention is to provide a osmanthus extract for the preparation of an antidepressant or health food. The therapeutically effective amount of the sweet-scented osmanthus extract can be used for the treatment of a drug or health food for treating or preventing sorrow. . A second object of the present invention is to provide a method for preparing an osmanthus extract to provide a osmanthus extract for use in the preparation of a medicament or health food for treating or preventing depression. Still another object of the invention is to provide a pharmaceutical composition for preventing or treating depression comprising a sweet-scented osmanthus extract and a pharmaceutically acceptable carrier or excipient. In order to achieve the foregoing object, the technical content of the present invention comprises: - an osmanthus extract, which is obtained by the following steps, an extraction step of immersing osmanthus in a solvent Forming a mixture-extracting mixture to extract (4) the antioxidant substance in the osmanthus flower to the extraction mixture=middle--filtering, removing the residue of the mixture (4) to obtain an extract; and The extract is subjected to a drying treatment to obtain a dried osmanthus extract. The osmanthus extract towel of the present invention, the osmanthus extract (4) is extracted from the flower part or dried flower part of Jingui, Yingui, Sijigui or Dangui. 1 201249450, two kinds of osmanthus extracts as described above, for the preparation of a medicament for preventing or treating sorrow or sedatives, the osmanthus extract has the performance of enhancing individual, antioxidant capacity and glutathione content, Preferably, the sweet-scented osmanthus extract is from the flowering department of Jingui, Yingui, Sijigui or Dangui. - a method for preparing an Osmanthus fragrans extract as described above, comprising: an extraction step of immersing osmanthus in a solvent to form an extractive mixture, and extracting the antioxidant material of the osmanthus fragrans from the solvent to the extraction mixture Second, 丄: over-age, remove the solid residue of the extraction mixture, two after two, two and a drying step 'take the extract to dry the place to obtain dried osmanthus extract. In the preparation method of the extract of Gui and Gui, the sweet-scented osmanthus is preferably a flower part or a dried flower part of Jinyingui, Sijiui or Dangui. The method for preparing the osmanthus extract of the present invention may have a weight ratio of 1:15 to 1:10. In the preparation method of the solvent, the solvent is preferably water, :. The preparation method of the ethyl, hexamethyst, the butyl hexazone, the hexanol, the hexagram, and the extract of the flower, the extraction step can be The preparation of the sweet-scented osmanthus extract of the present invention is dry = the evaporation treatment or the concentration method is used to treat the bacteria as a decompression method for preventing or treating the treatment (4) and the treatment amount - selected from the compound, Including: a soiled extract; and (b) 201249450 a pharmaceutically acceptable carrier or thief-shaped agent. - The pharmaceutical composition for preventing or treating a worry in the present invention, the medicine The pharmaceutical composition can be administered as a tablet, a capsule, a powder, a granule or a liquid. The pharmaceutical composition of the present invention for hiding or treating depression is more preferable. Preferably, the above-mentioned and other objects, features and advantages of the present invention are made more apparent and easy to understand. Example, sister's drawing, detailed The invention is as follows: Population The present invention relates to the use of a sweet-scented osmanthus extract in the preparation of anti-depressant drugs or health care. The therapeutically effective amount of sweet-scented osmanthus extract is used for the preparation of a medicament for treating or preventing (4) diseases, and the flower extract has Improve the antioxidant capacity of the brain and depression-related brain regions (such as the hippocampus) and promote blood circulation in the brain to help the brain resist free radicals or other oxides to achieve treatment or prevent depression. The sweet-scented osmanthus extract is taken from laurel (like var. thunbergiO, mk Q〇smanthus fragrans 胤lat god (6), four to i Osmanthus fragrans νΆτ. semperflorens, flutan 隹 (Osmanthus fragram eu aurantiacus), etc. The dried flower part is subjected to extraction of an antioxidant substance, and in this embodiment, the dried flower part of the golden laurel is taken for water extraction. Referring to FIG. 1 , the method for preparing the sweet-scented osmanthus of the present invention comprises an extraction step S1. a filtration step S2 and a drying step S3. 201249450 The extraction step si is performed by immersing osmanthus in a solvent to form an extraction mixture, and extracting the antioxidant substance in the osmanthus flower into the extraction mixture by the solvent. The solvent may be water, methanol, ethanol, acetone, ethane, propane, butane, hexane, petroleum ether or benzene. The weight ratio of osmanthus to the solvent is preferably 丨:15 to 丨:10. More specifically, the present embodiment uses water as a solvent, and after immersing the osmanthus in water for 15 minutes, the extraction mixture is boiled and extracted for at least 丨 hours to obtain the extraction. In the liquid mixture, the extraction step S1 is preferably repeated id times to completely extract the antioxidant substances in the sweet-scented osmanthus flower. In this embodiment, 9 8 kg of sweet-scented osmanthus is taken, extracted with 150 liters of boiling water for 丨 hours, and then immersed. After 丨 hours, the extracted osmanthus can be extracted again with 1 liter of boiling water to obtain a total of about 185 liters of the extract mixture. In this example, water is used as an extraction solvent, and the antioxidant of the broccoli f is extracted into the extract mixture to maintain a large amount of antioxidant substances in the extraction mixture. The filtration step S2 is to remove the solid residue of the extraction mixture to obtain an extract. The solid residue in the extraction mixture is removed, and the scale is taken to a volume suitable for sterilization by a vacuum concentration method, for example, high temperature and high __ to eliminate the extracted antioxidant substance. The drying step is magical. Thereby, it helps to preserve the anti-oxidation (four) f and the efficiency of the money. w = silk surface S3, take the fresh turn and carry on the treatment to obtain the sweet-scented osmanthus extract after 2. More specifically Yes, the Ganxian County can choose to treat it by processing, money-making, miscellaneous treatment or heat-drying treatment; this method can be used to concentrate and dry the antioxidant substances of sweet-scented osmanthus. The requirements are formulated into the required concentration for subsequent testing. To confirm the use of the sweet-scented osmanthus extract of the present invention in the preparation of the anti-worry (four) or health food, the osmanthus extract obtained by the above water extraction method is subjected to the test to carry out the test. The anti-oxygen (tetra) quality of the extract of Osmanthus fragrans L. (A) The extract of Osmanthus fragrans is used to improve the stagnation of the scent of the scented scent of the scent of the scented scent The total antioxidant capacity and gsh content of the 5th type |^ and (〇) hippocampus. (A) Antioxidant content test of sweet-scented osmanthus extract This example shows the total antioxidant capacity (〇xygen radical ^sorbance capacity, 〇RAC) and total phenolic content as the deduction of anti-oxidation lunar force. The total antioxidant capacity of the sweet-scented osmanthus extract was 18.8 ± 0.8 mM trolox equivalents, and the total phenolic content of the sweet-scented osmanthus extract was 65.7 ± 〇 77 mg GAE / ml '. The extract of the osmanthus extract of the present invention did contain antioxidant substances. (B) Osmanthus fragrans extract for improving the depression of the melancholic animal model (B1) Establishing a maternal deprivation animal model This example provides a melancholy animal model in the early isolation model (MaternaI deprivation model). Subsequent trials to suppress depression. The pregnant SD rats (SPF-class Sprageu-Dawley rat, referred to as SD rats purchased from Leko Biotech Co., Ltd.) were raised, and the first day after delivery of the pregnant SD rats was designated as the third day (pND〇). Among them, several male rats were born from the SD rats, and male SD rats of the experimental group and the control group were randomly selected. The SD rats in the control group (referred to as 201249450 CT gas) were co-raised in the normal way with the female rats; the sd rats in the experimental group (MD mice for short) were from PND2 to PND14, and the MD rats were isolated from the female rats for 1 hour every day. The male SD young a is placed in another space at 5 to 6 pm every day to prevent it from being disturbed by the external environment. After the MD rats were weaned (PND23), every four male SD rats were divided into one cage, and the temperature (22 lt:) and the light (12 small a temples) were maintained in the normal range. Environment, free and arrogant feed and water. Animal tests of t-type behavior can be performed until after PND 28. (B2) Animal test for melancholy behavior According to a study published by Porsolte/d. et al. (1977) in the journal Nature, 'the mouse can be reflected by the mouse in the case of Imm (Zhao~) Behavior shouting, that is, worrying about the coffee, special time to stay in the water for a long time can be used as a judgment of money _ degrees, the current basic medical research towel, water towel alone behavior as an important indicator of the mouse. According to the ladder (10) In the manner described, the class (4) rats were established for the test (= 2 financial experiment, Wei _) _ preparation - temperature suppression), before the test - the day needs to make the mice hot, the mice were placed in the sink 15 eight bubbles for 30 minutes For dry mice, gently with 60W, test, put the mouse into the same condition of the water tank 5 :: two into the water: do not move the struggle time within 5 minutes, from the calculation of the mouse, gp ^ r ^ 5lI, For 5 minutes, the time between the struggle and the time when the struggle is unsuccessful is taken as an indicator of depression. May Sen., 1st table, Taishen # /, group) and 6 groups of diligence (the second case is taken by a group of CT rats (Dijon, Knife (1) Bureaumate Bi to shame group
氣）進行桂花萃取物#、食實驗。 H又MD 〇中’第B〇&B1組係不 *一 10 201249450 儀食任何水或姑萃㈣，帛的__食純水，第B3 至B6紅分別係以每公斤體重之個體傲食〇 〇ι克、〇1克、 ^克及6克之桂花萃取物，該桂花萃取物較佳係以水溶解 後以8公分饒食官進行強迫管餘，且持續饒食14天後， 再進仃水巾不動試驗。本實施㈣進行三重覆試驗後取平 均值。Gas) osmanthus extract #, food experiment. H and MD 〇中'''''''''' The osmanthus extract of 〇〇g, 〇1g, ^g and 6g, the osmanthus extract is preferably dissolved in water and then forced to administer the tube with 8cm, and after 14 days of continuous feeding, Into the water towel does not move test. This implementation (4) takes the average value after the triple test.
一 v、也1亍'止蒂狀悲之U丄鼠，其 平均水中不動時間為Π5秒，第B1至B6組皆為類憂營鼠 (MD鼠）’其中’第B1及B2組的平均水中不動時間皆 大於I75秒，其憂營指數明顯較第B〇組高，然七，第抝 至B6組之MD鼠的水中不動時間皆與第別組並無顯 異（户<0力5)’表不服用每公斤體重Q ()1〜6 ()克桂花萃 之類憂鬱鼠，其憂#指數與對照組（第B〇經）相當 至比該對照組之憂營指數更低，Ί正實餵食本發明之桂w— 取物確實具有改善類憂鬱鼠的憂鬱情形之功效。土化卒 (C)雎部及各臟器之抗氧化能力試驗 在試驗(B)結束後犧牲各組別之老鼠，並採集其肝 脾臟、肺臟、腎臟及腦部。將腦部及各臟器以4。〇、、、 之 0.9% B4 B5 B6 0.1 1.0 6.0 131.6 61.2 55.5 201249450 生理食鹽水進行清理，哄私，、λ 之組㈣曾彳卜ji 乾私後稱重，將腦部及各臟器 之組織均Μ，H料㈣絲結軸 均質液，將該組織均質液Λ ^ 』、、且織 筲液以—組織均質緩衝液（Potassium ，PH7·4)定量至4倍體積後，置於鐵冷 康保存^騎後續(Cl)腦部及各臟器的總抗氧化力 (ORAC)f(C2)腦部及各臟器喊胱甘肽含量等試驗。 第2表所示係本實施例試驗(ci)之各組別 源’本貫施例之各組別係來自試驗⑻中第B2、组、第B5 組及第B6組的腦部、心臟、肝臟、脾臟、肺臟及腎臟之 組織均質液。 ^組織 組織來^^ 第B2组 第2, 腦部 C1-1 •試驗 心臟 C2-1 (ci)之— 肝臟 ----- C3-1 ‘組別 脾臟 C4-1 肺臟 C5-1 腎臟 C6-1 第B5組 C1-2 C2-2 C3-2 C4-2 C5-2 C6-2 第B6組 C1-3 C2-3 __C3-3__ C4-3 C5-3 C6-3 (C1)腦部及各臟器的總抗氧化力 由於憂營症的發生與腦部神經細胞受損有關，而腦部 神經細胞受損的·常常係來自自由基的攻擊，因此，藉 由測定腦部ϋ氧化力，以了解本發明之桂花萃取物用 於提升腦部抗氧化能力之效用。 腦部及各臟器的總抗氧化力（〇xygen radical absorba麟 capacity，簡稱 〇RAC)係根據 Chung #人(細4) 所建立之組織總氧化力測試方法加以修飾。 12 — 201249450 各組別之15μ1組織均質液分別與1〇〇μ1之藻紅素 (β-phycoerythrin ’ Ο.ΙμΜ)及 85μ1 之自由基誘導物（2, 2’-azobis(2-amidinopropane) dihydrochloride，簡稱 ΑΑΡΗ， 75mM)混合均勻，得一 ORAC測試液，再以波長為485nm 之光源激發該ORAC測試液，量測該ORAC測試液於波長 520nm之發散光’並以水溶性維生素E(Trolox)作為標準 品，當ORAC值為1，其相當於lmM水溶性維生素E之 抗氧化力。 請參照第3圖及第3表所示，第C1至C6組的總抗氧 化力白以CT乳為袁差，其次為儀食1.〇克/每公斤體重之 組別，最佳係餵食6.0克/每公斤體重之組別，其中，又以 弟C1組的腦部組織所提升之總抗氧化力表現最佳，特別 係第C1-1組與第C1-2組、第C1-3組皆具有顯著差異 (户<0.001)。 第3表：試驗(C)之ORAC值及GSH含量 組別 ORAC 值 GSH含量 (μΜ/mg protein ) C1-1 0.023 6.198 C1-2 1.807 15.661 C1-3 2.915 17.886 C2-1 2.013 6.429 C2-2 2.341 11.865 C2-3 3.552 13.952 C3-1 1.762 3.688 —13 — 201249450A v, also 1 亍 ' 止 状 悲 之 U U , , 其 其 , , , , , , , 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均 平均The immobility time in the water is greater than I75 seconds, and the sorrow index is significantly higher than that in the B group. However, the water in the third to the B6 group is not significantly different from the first group (household <0 force) 5) 'The table does not take every kilogram of body weight Q () 1 ~ 6 () grams of osmanthus extract and other melancholy rats, its worry # index and the control group (B 〇 经 经) is comparable to the control group's worry index The 桂 实 喂 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本Earthworms (C) Antioxidant capacity test of the ankle and various organs At the end of the test (B), rats of each group were sacrificed, and the liver, spleen, lungs, kidneys and brain were collected. Take 4 brains and organs. 0.9% of 〇,,和, B4 B5 B6 0.1 1.0 6.0 131.6 61.2 55.5 201249450 Physiological saline is cleaned, smuggled, λ group (four) Zeng 彳 ji dry and then weighed, the organization of the brain and organs Uniform, H material (four) silky shaft homogenization liquid, the tissue homogenization liquid Λ ^ 』, and the woven liquid is quantified to 4 times volume with tissue homogenization buffer (Potassium, PH7·4), placed in iron cold Kang preserves ^ rides follow-up (Cl) brain and various organs of total antioxidant capacity (ORAC) f (C2) brain and various organs called glutathione content and other tests. Table 2 shows the respective groups of the test (ci) of this example. The respective groups of the present examples are from the brain, heart, and the B2, B5, and B6 groups in the test (8). The tissues of the liver, spleen, lungs and kidneys are homogeneous. ^Organizational organization^^ Group B2 2nd, Brain C1-1 • Test heart C2-1 (ci) - Liver----- C3-1 'Group spleen C4-1 Lung C5-1 Kidney C6 -1 Group B5 C1-2 C2-2 C3-2 C4-2 C5-2 C6-2 Group B6 C1-3 C2-3 __C3-3__ C4-3 C5-3 C6-3 (C1) Brain and The total antioxidant capacity of each organ is related to the damage of nerve cells in the brain due to the occurrence of stagnation, and the damage of nerve cells in the brain is often caused by free radical attack. Therefore, by measuring the oxidative power of the brain To understand the effect of the osmanthus extract of the present invention for enhancing the antioxidant capacity of the brain. The total antioxidant capacity of the brain and various organs (〇xygen radical absorba lin capacity, referred to as 〇RAC) was modified according to the total oxidative strength test method established by Chung #人(细4). 12 — 201249450 15μ1 tissue homogenization solution of each group with 1〇〇μ1 phycoerythrin (β-phycoerythrin 'Ο.ΙμΜ) and 85μ1 of free radical inducer (2, 2'-azobis(2-amidinopropane) dihydrochloride , abbreviated as ΑΑΡΗ, 75 mM), evenly mixed, an ORAC test solution was obtained, and the ORAC test solution was excited by a light source having a wavelength of 485 nm, and the ORAC test solution was measured for divergence at a wavelength of 520 nm and water-soluble vitamin E (Trolox) was used. As a standard, when the ORAC value is 1, it is equivalent to the antioxidant power of lmM water-soluble vitamin E. Please refer to the 3rd and 3rd tables. The total antioxidant power of the C1 to C6 groups is the difference of the CT milk, followed by the group of 1. gram/kg body weight. In the group of 6.0 g/kg body weight, the total antioxidant capacity of the brain tissue of the brother C1 group was the best, especially the C1-1 group and the C1-2 group, the C1-3 group. Groups were significantly different (household < 0.001). Table 3: ORAC value and GSH content of test (C) ORAC value GSH content (μΜ/mg protein) C1-1 0.023 6.198 C1-2 1.807 15.661 C1-3 2.915 17.886 C2-1 2.013 6.429 C2-2 2.341 11.865 C2-3 3.552 13.952 C3-1 1.762 3.688 —13 — 201249450
由試驗(ci)可知，本發明之桂花萃取物 腦部及各臟器之總抗氧化力具有明顯之效用， 升腦部之總抗氧化力有明顯的效果。 (C2)腦部及各臟器的麵胱甘肽含量 對於提升個體 且特別係提 由於麩胱甘肽（Glutathione，簡稱GSH)係具有清除 自由基以保護腦部神經細胞之效用，因此，藉由測量本發 明之桂花萃取物提升腦部神經細胞之GSH含量，證實本發 明之桂花萃取物係能夠保護腦部神經細胞而達到抗憂鬱之 效果。 試驗(C2)測量GSH係根據Anderson(1985)所提出之方 法加以修飾。該GSH之測量原理係利用GSH能夠將一指 示劑（5-5’-dithiobis(2-nitrobenzoicacid)，簡稱 DTNB)還 原成2nitro-5-benzoic (簡稱TNB)，以分光光度計測量各 201249450 組別之TNB於波長為405nm處之吸光值，藉以判斷各組 別之GSH含量。 本實施例係取如第2表所示，各組別ι5〇μ1之組織均 質液分別加入450μ1之5%三氯醋酸溶液（簡稱tca)均 勻混合’付一 TCA如處理液’以l〇〇〇〇rpm離心分鐘， 取30μ1之TCA前處理液與140μ1之TRIS緩衝液及10μ1 之DTNB (0.01Μ)混合後靜置5分鐘後，測定各組別於 405nm下之吸光值。 請參照第4圖及第3表所示，第C1至C6組的GHS 含量皆以CT鼠為最少，其次為傲食1〇克/每公斤體重之 組別，隶佳係儀食6.0克/每公斤體重之組別，其中，又以 第C1至C3組的腦部、心臟及肝臟等組織中的GHS增加 量最多’特別係第C1-1組分別與第Cl_2及C1_3組皆具有 顯著差異（/?<0.00〇。 由試驗(C2)可知，本發明之桂花萃取物對於提升個體 腦部及各臟器之GSH含量具有明顯之效用，且特別係提升 腦部之GSH含量有明顯的效果。如此，係有助於改善類憂 鬱鼠之憂鬱情形。 (D)海馬迴區之總抗氧化力及GSH含量 反誊症與細部中各腦區之抗氧化能力衰退有關，其 中，又以海馬迴區與憂鬱症之關係最為密切，根據Wu等 人於 2008 年發表之研究（Behavi〇ural Brain Research (193) 183-191)指出，一旦海馬迴區之抗氧化能力變弱或gSH 含量減少，則發生憂鬱症之情況將會大大提升。 。月參如、第4表所示，本實施例之各組別係來自試驗(b) —15 — 201249450 中第B2至B5組的MD鼠腦部中的海馬迴區，其中，本實 施例之第D1至D4組之海馬迴組織係來自於試驗(B)中未 儘食任何水或桂花萃取物的MD鼠為第D1組，而第D2 至D3組則分別為餵食0.01克、0.1克及1克桂花萃取物/ 每公斤體重之MD鼠，並以試驗(C1)及(C2)所述之方法分 別測量第D1至D4組之總抗氧化力及GSH含量。 凊參照第5及6圖所示’第D1至D4組的總抗氧化力 及GSH含量皆以第D1組為最少’其次依序為第D2、D3 及D4組，第D2至D4組與第D1組之間具有顯著差異， 其中，又以每公斤體重餵食〇.1克及1.0克之組別所測得之 總抗氧化力為最高。 f 4表：試驗⑼之〇RAC值及GSH含量 組別 ORAC 值 GSH含量 (μΜ/mg protein) D1 0.034 0.857 D2 0.197 3.690 D3 0.873 4.320 D4 0.913 5.700 由試驗(D)可知，本發明之桂花萃取物對於提升個體腦 部海馬迴區之總抗氧化力及GSH含量具有_之制，該· 桂花萃取物確實係透過保護海馬迴區之神經細胞以達到抗 憂鬱之功效。 本土月之桂彳b萃取物係能夠提升與憂相關腦區之 抗氧化能力’幫助腦部神經細胞抵抗自由基或其他氧化物 一 16 — 201249450 之侵襲，將-治療或預防有效劑量之桂花萃取物用於製備 &療，預防憂鬱症之樂物或保健食品，特別係與醫藥學上 可接叉之細或卿劑組合形成—醫藥組合物，其中，該 桂花萃取物係可以製備成任何方便食用之型式，如錠劑、/ 膠囊、粉劑、_或液料，或者將該桂花萃取物與其他 食品或飲料組合’以—適於食狀樣態供生物體以口服方 j服用。該醫藥組合物巾’其桂花萃取物之㈣較佳係每 單位體重之個體投予0.01至6,〇克/公斤體重，且投予時間 可持續14天’具有達脱療或驗憂之功效。 本發明之桂花萃取物在製備抗㈣藥物或保健食品 之用途’-治療有效劑量之桂花萃取物係可用於製備治療 或預防憂鬱症之藥物或保健食品，具有提供憂鬱症患者— 種新的抗憂鬱藥物或保健食品之功效。 —本發明桂花萃取物之製備方法，係用以提供一種桂花 萃取物以製備治療或預防錄症之藥物或保健食品，具 增進桂花之經濟價值之功效。 八 本發明之桂花萃取物在製備抗憂t#物或保健食品 之用途’其係可與醫藥學上可接受之侧或卿劑組合形 成-醫藥組合物，㈣治療憂#症之麵，具有改善 症患者憂鬱情形之功效。 ^ 、，本發明已_上述較佳實施例揭示，然其並非用 以，疋本剌’任何熟纽技藝者在本發明之精神 1乾圍之内’相對上述實施例進行各種更動與修改仍屬本 X月所保4之技術範·，因此本發明之保護範圍當視後附 之申請專利範圍所界定者為準。 17 — 201249450 【圖式簡單說明1 第1圖：本發明萃取桂花之製備方法的步驟方塊圖。 第2圖：本實施例水中不動試驗之長條圖。 第3圖：本實施例腦部及各臟器之總抗氧化力長條圖。 第4圖：本實施例腦部及各臟器之GSh含量長條圖。 第5圖：本實施例海馬迴區之總抗氧化力長條圖。 第6圖：本實施例海馬迴區之GSH含量長條圖。 【主要元件符號說明】 51 萃取步驟 52 過濾步驟 53 乾燥步驟 18 —It can be seen from the test (ci) that the total antioxidant power of the osmanthus extract of the present invention in the brain and various organs has a significant effect, and the total antioxidant capacity of the ascending brain has a significant effect. (C2) The content of caspase in the brain and various organs is important for enhancing the individual and especially because the glutathione (GSH) system has the effect of scavenging free radicals to protect brain nerve cells. The GSH content of brain cells was measured by measuring the osmanthus extract of the present invention, and it was confirmed that the osmanthus extract of the present invention can protect brain nerve cells and achieve anti-depression effect. Test (C2) Measurement GSH was modified according to the method proposed by Anderson (1985). The GSH measurement principle uses GSH to reduce an indicator (5-5'-dithiobis (2-nitrobenzoic acid), DTNB for short) to 2nitro-5-benzoic (TNB for short), and to measure each group of 201249450 by spectrophotometer. The TNB has an absorbance at a wavelength of 405 nm to determine the GSH content of each group. In this embodiment, as shown in the second table, the tissue homogenization liquid of each group ι5〇μ1 is separately added with 450 μl of 5% trichloroacetic acid solution (abbreviated as tca), and uniformly mixed with 'paying a TCA such as a treatment liquid' to After centrifugation at rpm for rpm, 30 μl of the TCA pretreatment solution was mixed with 140 μl of TRIS buffer and 10 μl of DTNB (0.01 Å), and allowed to stand for 5 minutes, and then the absorbance at 405 nm of each group was measured. Please refer to Figure 4 and Table 3. The GHS content of Groups C1 to C6 is the least for CT rats, followed by the group of 1 gram per kilogram of body weight. In the group of kilograms of body weight, the increase in GHS in the brain, heart and liver tissues of the C1 to C3 groups was the highest. In particular, the C1-1 group had significant differences with the Cl_2 and C1_3 groups, respectively. (/?<0.00〇. From the test (C2), the sweet-scented osmanthus extract of the present invention has obvious effects for improving the GSH content of the brain and various organs of the individual, and particularly improves the GSH content of the brain. The effect is to improve the depression of the melancholic rats. (D) The total antioxidant capacity and GSH content in the hippocampus area are related to the decline of antioxidant capacity in each brain region in detail. The hippocampus area is most closely related to depression. According to a study published by Wu et al. in 2008 (Behavi〇ural Brain Research (193) 183-191), once the hippocampal area has weakened antioxidant capacity or reduced gSH content, , the situation of depression will be greatly improved. As shown in the fourth table, each group of the present embodiment is the hippocampal gyrus in the brain of the MD mouse of the group B2 to B5 of the test (b)-15-201249450, wherein the D1 to D4 of the present embodiment The group of hippocampus was from the D1 group in the test (B), and the D2 to D3 groups were fed 0.01 g, 0.1 g and 1 g of sweet-scented osmanthus extract. MD/kg/kg body weight, and the total antioxidant capacity and GSH content of groups D1 to D4 were measured by the methods described in tests (C1) and (C2). 凊 Refer to Figures 5 and 6 The total antioxidant capacity and GSH content in the D1 to D4 groups were the least in the D1 group, followed by the D2, D3 and D4 groups. There was a significant difference between the D2 and D4 groups and the D1 group. The total antioxidant capacity measured by the group of 1 g and 1.0 g per kg of body weight was the highest. f 4 Table: RAC value and GSH content after test (9) ORAC value GSH content (μΜ/mg protein) D1 0.034 0.857 D2 0.197 3.690 D3 0.873 4.320 D4 0.913 5.700 From the test (D), the sweet-scented osmanthus extract of the present invention is used to enhance the hippocampal area of the individual brain. The total antioxidant capacity and GSH content have a system of _ scented osmanthus extracts through the protection of nerve cells in the hippocampus to achieve anti-depression effect. The local laurel b extract system can enhance the brain area related to worry Its antioxidant capacity 'helps brain nerve cells fight free radicals or other oxides. — 201249450 Invasion, will treat or prevent effective doses of osmanthus extract for preparation & treatment, prevention of depression or health care The food product, in particular, is formed by combining a pharmaceutically acceptable fine or a fine agent to form a pharmaceutical composition, wherein the sweet-scented osmanthus extract can be prepared into any convenient form such as a tablet, a capsule, a powder, or The liquid material, or the osmanthus extract is combined with other foods or beverages, to be taken in an edible form for the organism to be taken orally. The pharmaceutical composition towel 'the osmanthus extract (4) is preferably administered to an individual per unit weight of 0.01 to 6, gram per kilogram of body weight, and the administration time can last for 14 days, which has the effect of achieving de-therapy or healing. . The use of the sweet-scented osmanthus extract of the present invention in the preparation of anti-(four) drugs or health foods--the therapeutically effective amount of sweet-scented osmanthus extracts can be used for preparing medicines or health foods for treating or preventing depression, and providing patients with depression - a new kind of anti-drug The efficacy of depression drugs or health foods. The method for preparing an osmanthus extract of the present invention is for providing a sweet-scented osmanthus extract to prepare a medicine for treating or preventing a disease or a health food, which has the effect of improving the economic value of sweet-scented osmanthus. The use of the eight-invention osmanthus extract in the preparation of anti-worry t# or health foods can be formed in combination with a pharmaceutically acceptable side or a medicinal composition - a pharmaceutical composition, (4) a therapeutic remedy, having Improve the efficacy of depression in patients with symptoms. The invention has been disclosed in the above preferred embodiments, but it is not intended to be used in the context of the spirit of the present invention. It is the technical scope of the 4th edition of this X-month. Therefore, the scope of protection of the present invention is subject to the definition of the scope of the patent application. 17 — 201249450 [Simplified illustration of the drawing 1 Fig. 1 is a block diagram showing the steps of preparing the sweet-scented osmanthus of the present invention. Figure 2: Long strip diagram of the immobile test in this example. Fig. 3 is a bar graph showing the total antioxidant power of the brain and organs of the present embodiment. Figure 4: Bar graph of GSh content in the brain and organs of this embodiment. Fig. 5 is a bar graph showing the total antioxidant power of the hippocampus in the present embodiment. Fig. 6 is a bar graph showing the GSH content of the hippocampus in the present embodiment. [Main component symbol description] 51 Extraction step 52 Filtration step 53 Drying step 18 —