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TWI258010B - Medicament and antihyperglycemic effects of resveratrol - Google Patents

Medicament and antihyperglycemic effects of resveratrol Download PDF

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TWI258010B
TWI258010B TW93114821A TW93114821A TWI258010B TW I258010 B TWI258010 B TW I258010B TW 93114821 A TW93114821 A TW 93114821A TW 93114821 A TW93114821 A TW 93114821A TW I258010 B TWI258010 B TW I258010B
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Taiwan
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resveratrol
insulin
rats
dependent
dose
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TW93114821A
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Chinese (zh)
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TW200420882A (en
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Li-Man Hung
Jan-Kan Chen
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Univ Chang Gung
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Abstract

Resveratrol, an antioxidant found in various fruits, vegetables, grapes, and red wines, has been shown to have free radical scavenging, antiflammatory and antithrombogenic activities in the biological systems. Moreover, resveratrol has also been shown to be a good cancer preventive chemical and has been recommended as a food supplement for health benefits. Recently, we found that resveratrol can effectively lower the blood sugar in both type I (insulin dependent) and type II (insulin independent) diabetic rats. Resveratrol could also delay the onset of the insulin-resistant phenotype in the experimental rats with prolonged injection of insulin. The hypoglycemic effect in conjunction with its aforementioned health benefits, favors the use of resveratrol as a health food component and as a therapeutic drug to treat patients with hyperglycemic conditions.

Description

1258010 玫、發明說明: 【發明所屬之技術領域】 本發明係為一種證明白黎蘆醇具有降血糖藥效的方法及其藥劑,尤指一 種證明白黎蘆醇對胰島素依賴型(IDDM)與胰島素非依賴型(NIDDM)糖尿病皆 具療效’並可延緩胰島素耐受性之產生之醫藥技術領域。 【先前技術】 糖尿病的病變描述已有二千年以上的歷史,在「内經素問」及「金置要 略載」已有相關的記載描述。糖尿病的古名為「消渴症」,主要是联臟所分 泌的胰島素之量不足,而血中的糖分增加,致糖分由尿中排出;特有症狀為 口渴、多尿、倦怠、人漸消瘦等。估計於2000年全世界約有一億三千萬人 口罹患糖尿病,其中約5—10 %為第一型(胰島素依賴型)糖尿病而9〇-95% 為第一型(胰島素非依賴型)糖尿病。糖尿病病患的人數在我國亦有逐漸增 加之趨勢,估計全台灣約有90萬糖尿病人口。依據1998年的健保資料分析, 國内總共有53· 6萬接受藥物治療的糖尿病人,且糖尿病也連續五年居本國 十大死亡病因之一。事實上,糖尿病之罹病比率雖併列在心血管疾病之側, 可是,糖尿病治療藥物的種類卻遠不如降血壓藥、心絞痛治療藥或中風治療 藥之多。當然,騰島素能有效的控制血糖,也阻擾了這類藥物的開發。然而, 糖尿病的病變裡,有許多仍是騰島素所無法控制的,加上長期注射胰島素所 出現的胰島素耐受性(insulin resistance)現象料,都是促使醫學界渴 望研發其他有效降血糖治療藥物之原因。 目前,糖尿病的病人常被歸納為「騰島素依賴型(麵)」和「騰島素 非依賴型(NIDDM)」二大類,治療方式也因此而異。常㈣口服降血糖藥物 略分為禮__和biguanides二大類,均為配合肤島素控制病人血 糖的藥物。Sulf〇nyl_類的藥物藉由刺激體内細胞分泌膜島素,故適合 於_M病人的血糖控制,對於沒辦法分泌騰島素的臓病人則無療效。 1258010 biguanides藥物則是以改善姨島素促進周邊組織對葡萄糖利用的效率為 主,並不刺激胰島素的分泌。 【發明内容】 本發明之主要目的係在提供一種證明白黎蘆醇具有降血糖藥效的 方法及其藥劑,尤指一種證明白黎蘆醇對胰島素依賴型(IDDM)與胰島素 非依賴型(NIDDM)糖尿病皆具療效。並可延緩胰島素财受性之產生且有控制 體重之效果。 為達前述之目的,本發明提供了 一種證明白黎廑醇具有降血糖藥效 的方法及其藥劑,糖尿病可怕之處在於經年累月的高血糖容易併發其他疾春 病,如特異性病變如視網膜病變、腎臟病變及神經病變,及非特異性病變如 中風、心臟病、下肢血管的硬化、白内障等,造成病人的生命威脅。因此治 療糖尿病之藥物除了應具有降血糖之作用外、若能同時兼具減少糖尿病併發 症之產生作用則更為理想。而反式白黎蘆醇,又名反式三羥基芪 (Trabs-resveratrol),是葡萄等植物的次生代謝産物,具有抗氧化、掃除 自由基、抗炎症、降低血脂肪、防止血栓、使冠狀動脈血栓造成的心臟病的 死亡率降低等作用,在癌細胞的啓始、促進及擴展的三個階段中均具有化學 防癌活性。因此,美國的癌症研究專家已經向人們提出防癌新建議:多吃葡鲁 萄,不吐葡萄皮。並且認爲,一般的葡萄酒飲酒量已可達到白黎蘆醇的有效 量。若將白黎蘆醇resveratrol配製成不同劑型的飲品或精製成藥,可爲糖 尿患者找到另外一種有效降血糖途徑。本成果將具有巨大的醫療經濟利益。 故白黎蘆醇resveratrol適合做為健康食品的添加物,及中、西藥廠開發為 臨床降血糖藥物。 惟上述之說明係僅為本發明部分特徵之概述,為使貴審查委員及閱讀 本說明書之專業人士,皆能更明白了解本發明之技術手段,並可依本說明書 内容據以實施’故對本發明配合圖不加以說明,以下所述僅為用以解釋本發 6 125801ο 日月車交4圭官- 、 細*例’並非對本發明做任何形式上之限制,是以,凡在相同之創 月神下所作有關本發明之任何修飾或變更,皆應包括在本發明保護之範 圍。 【實施方式】 本發明之實驗方法依序為: 1實驗動物來源:構自國家動物中心的Sprage-Dawley (SD)品系的 雄陡大白鼠,年齡在1 2 3 4 5 6週以上,體重約200-250g之間。飼養於長庚醫院動 物中心以空調維持室溫在25±1。(:的動物室,光照時間與黑暗時間各為12小 時’讓老鼠自由的進食與飲水。 2給藥的方法:白黎蘆醇resveratrol (購自Sigma)溶於生理食鹽水後, 以〇·〇5、〇·ι、05 mg/ml之量,再經由給藥管灌食到不同型的糖尿病老鼠的 胃邛,依動物的體重的差異,灌食不同容量的藥液以達到需要的濃度 (mg/kg) 〇 1 誘導大白鼠產生胰島素依賴型(Iddm)糖尿病:將8週的SD雄 f生大白鼠禁食72小時後,以腹腔注射pent〇barbital 30 mg/kg將之麻醉,自 舨靜脈注射,給予老鼠streptozotocin ( STZ, 65 mg/kg)。於72小時後,抽取 靜脈血,利用葡萄糖試劑(glucose Kit),經由自動分析儀(Autoanalyser, Quik-Lab” Ames)得知血糖值。當老鼠的血糖值大於400 mg/(JL,並有糖尿病 二多症狀出現(多吃、多喝、多尿),即視為胰島素依賴型(IDDM)糖尿病老鼠。 2 誘導大白鼠產生非胰島素依賴型(NIDDM)糖尿病,其中又分為兩 3 點: 4 (1 )胰島素耐受性型(NIDDM-insulin):利用SD系的雄性大白鼠, 週齡8週大,以皮下注射給予長效型的胰島素(M〇n〇tard(g) HM;N〇v() Nwjisk, 51258010 玫,发明说明: [Technical Field of the Invention] The present invention is a method for demonstrating the efficacy of resveratrol with hypoglycemic agents and an agent thereof, and more particularly to demonstrate that resveratrol is insulin-dependent (IDDM) and Insulin-independent (NIDDM) diabetes has therapeutic efficacy and can delay the development of insulin resistance in the field of medical technology. [Prior Art] The description of the lesions of diabetes has a history of more than 2,000 years. It has been described in the "Internal Sutra" and "Golden Sets". The ancient name of diabetes is "diabetes". The main reason is that the amount of insulin secreted by the joint organs is insufficient, and the sugar in the blood is increased, and the sugar is discharged from the urine. The special symptoms are thirst, polyuria, burnout, and people gradually. Thin and so on. It is estimated that about 1300 million people worldwide suffer from diabetes in 2000, of which about 5-10% are type 1 (insulin-dependent) diabetes and 9〇-95% are type 1 (insulin-independent) diabetes. . The number of people with diabetes is also increasing in China. It is estimated that there are about 900,000 people with diabetes in Taiwan. According to the analysis of health care data in 1998, there were a total of 536,000 diabetic patients receiving drug treatment in China, and diabetes has also been among the top ten causes of death in the country for five consecutive years. In fact, the rickets rate of diabetes is listed on the side of cardiovascular disease. However, the types of diabetes drugs are far less than those of blood pressure lowering drugs, angina pectoris drugs or stroke therapeutic drugs. Of course, Tengdao can effectively control blood sugar and also hinder the development of such drugs. However, many of the lesions of diabetes are still beyond the control of Tengdaosu, and the insulin resistance phenomenon caused by long-term injection of insulin is urging the medical community to develop other effective hypoglycemic treatments. The cause of the drug. At present, patients with diabetes are often classified into two categories: "Tengdaosu-dependent (face)" and "Tengdaosu-independent (NIDDM)", and the treatment methods vary. Often (four) oral hypoglycemic drugs are divided into two categories of ritual __ and biguanides, all of which are combined with the control of the patient's blood sugar. The Sulf〇nyl® drug is suitable for the glycemic control of _M patients by stimulating the secretion of membranein in the cells of the body, and is ineffective for patients with sputum who cannot secrete tamsin. 1258010 The biguanides drug is based on improving the efficiency of gingivalin in promoting the utilization of glucose by surrounding tissues, and does not stimulate insulin secretion. SUMMARY OF THE INVENTION The main object of the present invention is to provide a method for demonstrating the efficacy of resveratrol with hypoglycemic agents and an agent thereof, and more particularly to demonstrate that resveratrol is insulin-dependent (IDDM) and insulin-independent ( NIDDM) Diabetes is effective. It can also delay the production of insulin and has the effect of controlling body weight. For the purpose of the foregoing, the present invention provides a method and a medicament for demonstrating the efficacy of resveratrol with hypoglycemic agents, and the horrible condition of diabetes is that hyperglycemia over the years is easy to be complicated by other diseases such as retinopathy. Renal lesions and neuropathy, as well as non-specific lesions such as stroke, heart disease, hardening of lower extremity blood vessels, cataracts, etc., cause life threats to patients. Therefore, in addition to the effect of lowering blood sugar, the drug for treating diabetes is more desirable if it can simultaneously reduce the incidence of diabetes mellitus. Trans-resveratrol, also known as trans-trishydroxypyrene (Trabs-resveratrol), is a secondary metabolite of plants such as grapes. It has anti-oxidation, eliminates free radicals, resists inflammation, lowers blood fat, prevents blood clots, and makes The mortality of heart disease caused by coronary thrombosis is reduced, and it has chemical anti-cancer activity in the three stages of initiation, promotion and expansion of cancer cells. Therefore, cancer research experts in the United States have already proposed new anti-cancer recommendations: eat more Portuguese and do not spit grape skin. It is also believed that the average amount of wine consumed can reach the effective amount of resveratrol. If resveratrol resveratrol is formulated into a different type of drink or refined drug, another effective blood glucose lowering route can be found for diabetic patients. This achievement will have enormous medical and economic benefits. Therefore, resveratrol resveratrol is suitable as an additive for health foods, and is developed as a clinical hypoglycemic drug by Chinese and Western medicine factories. However, the above description is only an overview of some of the features of the present invention, and the technical means of the present invention can be more clearly understood by the reviewing committee and the professionals who read the present specification, and can be implemented according to the contents of the present specification. The invention is not described in the following figures. The following description is only used to explain the fact that the present invention is not limited to any form of the invention. Any modifications or variations of the present invention made by Luna should be included in the scope of the present invention. [Examples] The experimental methods of the present invention are in order: 1 Experimental animal source: Male and steep rats of the Sprage-Dawley (SD) strain constructed from the National Animal Center, aged 1 2 3 4 5 6 weeks or more, weighing about Between 200-250g. The animal center in Chang Gung Memorial Hospital was maintained at room temperature at 25±1 with air conditioning. (: animal room, light time and dark time are each 12 hours 'allow the rats to eat and drink freely. 2 Method of administration: resveratrol resveratrol (purchased from Sigma) dissolved in physiological saline, 〇5, 〇·ι, 05 mg/ml, and then fed to the stomach cramps of different types of diabetic rats via the drug tube. According to the difference in body weight, different doses of liquid are administered to achieve the desired concentration. (mg/kg) 〇1 induces insulin-dependent (Iddm) diabetes in rats: after 8 weeks of SD male f-white rats were fasted for 72 hours, they were anesthetized by intraperitoneal injection of pent〇barbital 30 mg/kg. After intravenous injection, rats were given streptozotocin (STZ, 65 mg/kg). After 72 hours, venous blood was drawn and the blood glucose level was determined by an automatic analyzer (Autoanalyser, Quik-Lab" Ames) using a glucose kit. When the mouse's blood sugar level is greater than 400 mg / (JL, and there are more than two symptoms of diabetes (eat more, drink more, more urine), it is considered as insulin-dependent (IDDM) diabetic mice. 2 induced non-insulin production in rats Dependent (NIDDM) diabetes, of which Divided into two points: 4 (1) Insulin-tolerant (NIDDM-insulin): male SD rats using SD, 8 weeks old, subcutaneous injection of long-acting insulin (M〇n〇tard) (g) HM; N〇v() Nwjisk, 5

Bagsvaerd,Denmark),每天投予三次,每次劑量〇·5 iu/kg,持續兩個星期後, 6 會產生高血糖。利用tolbutamide,來加以確定老鼠已經產生胰島素耐受性 1258010 (resistance)的現象,即視為非胰島素依賴型(NIDdM)糖尿病老鼠。 (2 )肥胖型(NIDDM-nicotinamide) : SD雄性大白鼠禁食48小時後, 以 pentobarbital 30 mg/kg 麻醉後,先腹腔注射 nicotinamide 1〇〇 mg/kg 後再由 股靜脈注射streptozotocin( STZ,65 mg/kg)。於四周後,抽取靜脈血,利用葡 萄糖試劑(glucose Kit),經由自動分析儀(Autoanalyser,Quik心b,Ames)得 知血糖值。當老鼠的血糖值大約等於200 mg/dL,並有肥胖現象,即視為非 胰島素依賴型(NIDDM)糖尿病老鼠。Bagsvaerd, Denmark), administered three times a day, each dose 〇·5 iu/kg, after two weeks, 6 will produce hyperglycemia. Tolbutamide was used to determine that the mouse has developed insulin resistance 1258010 (resistance), which is considered to be a non-insulin dependent (NIDdM) diabetic mouse. (2) Obesity (NIDDM-nicotinamide): After 48 hours of fasting in SD male rats, after anesthesia with pentobarbital 30 mg/kg, intraperitoneal injection of nicotinamide 1〇〇mg/kg followed by intravenous injection of streptozotocin (STZ, 65 mg/kg). After four weeks, venous blood was drawn, and the blood glucose level was obtained by an automatic analyzer (Autoanalyser, Quik heart b, Ames) using a glucose kit. When a mouse has a blood glucose level of approximately 200 mg/dL and is obese, it is considered a non-insulin-dependent (NIDDM) diabetic mouse.

5、 血糖的檢測方法:首先,在實驗前一天晚上,先將SD老鼠、STZ 或NIDDM的糖尿病老鼠禁食。隔天,將老鼠麻醉以腹腔注射pent〇barbital3〇鲁 mg/kg。先抽空腹血糖〇·2-〇·6 ml之後,再以灌食給予白黎遂醇resveratr〇1 於老鼠的體内。60、90、120分鐘後,再抽血來進行離心,(13 〇〇〇rpm,3分 鐘)以分離血清和血漿。取上層的血清1〇 ul,加入gluc〇se kit反應試劑i 〇 ml,混合均勻後,於37°C恆溫水槽中反應10分鐘。接著利用血糖機 (Quick-Lab,Chemistry Analyzer)對其吸光度的不同來測血糖值。Gluc〇se kit反應試劑的原理乃利用葡萄糖氧化成葡萄糖酸和過氧化氫,藉著葡萄糖 的色素基氧化呈現深淺不同的紅色。血糖值乃由血清和gluc〇se kit反應所呈 現的紅色;顏色變化可由吸光度差異感知,並與標準品對照換算出血糖值鲁 (mg/dL) ° 6、 統計分析:實驗所得的結果以平均值±標準誤差(mean±SE)來表示, 再以Student’s t-test來評估兩者間的差異,多組比較時則使用AN〇VA和 Dunnett’s Post-hoc test來評估之間的差異。p<〇 〇5,即視為有顯著的差異。 而本發明之實驗結果如下: 1、白黎蘆醇resveratrol對正常大白鼠(SD)之降血糖作用:請配合參 照第一圖所示,給予SD大白鼠口服不同劑量的白黎蘆醇resvemtr〇1 (〇 〇5、 0·1、0.5 mg/kg)。經過90分鐘後’發現白黎蘆醇resveratr〇i隨著劑量的增加, 8 1258010 可以明顯地對SD大白鼠產生降血糖的作用。灌食白黎蘆醇resveratr〇1 〇1 mg/kg所產生的降血糖作用約25·72±4·03%。於0.5 mg/kg的劑量下,白黎蘆 醇resveratrol所產生降血糖的作用可達到最大,約35.53±2.0%。 2、 白黎蘆醇resveratrol對胰島素依賴型(IDDM)大白鼠之降血糖作 用:請配合參照第二圖所示,給予IDDM大白鼠口服不同劑量的白黎麓醇 resveratrol ( 0·05、0·1、0·5 mg/kg)。經過 90 分鐘後,發現白黎蘆醇 resveratr〇i 隨著劑量的增加,可以明顯地對IDDM老鼠產生降血糖的作用。灌食白黎蘆 醇 resveratrol 0.1 mg/kg 所產生的降血糖作用約 21·00±5.84%。於 0.5 mg/kg 的劑量下,白黎釐醇resveratrol所產生降血糖的作用可達到最大,約23〇〇土 1.89%。 3、 白黎蘆醇resveratrol對非胰島素依賴型(NIDDM)大白鼠之降血糖 作用:請配合參照第三圖及第四圖所示,給予NIDDM大白鼠口服不同劑量 的白黎蘆醇resveratrol (0·05、0.1、0.5 mg/kg)。經過90分鐘後,發現白黎 蘆醇resveratrol隨著劑量的增加,可以明顯地對NIDDM老鼠產生降血糖的 作用。對騰島素财受性型-NIDDM灌食白黎逢醇resveratrol 0.1 mg/kg所產生 的降血糖作用可達28·3±1·67%。並且,在〇·5 mg/kg的劑量下,白黎蘆醇 resveratrol所產生降血糖的作用可達到最大,約32.UL62%。對肥胖型 -NIDDM灌食白黎JI醇resveratr〇l 〇·ι mg/kg所產生的降血糖作用可達26.43 ±2.48% 〇 4、 比較口服白黎蘆醇resveratr〇l後對正常、IDDM、NIDDM大白鼠之 降血糖作用:請配合參照第五圖所示,口服白黎蘆醇resveratrol 0.5 mg/kg 90 分鐘後所產生的降血糖作用,在有胰島素存在的正常、NIDDM-insulin及 NIDDM-nicotinamide大白鼠,所產生降血糖的百分比分別為35.53±2.0%、 32·1±1·62%、24·86±1·32% (N=8),在無胰島素的IDDM型大白鼠產生的降 血糖百分比則為23·00±1·89%(Ν=8)。 1258010 5、白黎蘆醇⑽veratrol延緩SD大白鼠產生胰島素耐受性··請配合參 照第六圖所示,每天投予正常SD大白鼠胰島素(〇.5lu/kg,Tid)時,大白鼠於 16·5 ± 1.5天時開始產生腺島素耐受性(insuHn resistance),而白黎蘆醇 resvemtrol (0·5 mg/kg,Bid)治療組有效地延遲胰島素耐受性之產生、於23 5士 1.5天時動物才產生胰島素耐受性。同時我們亦發現白黎蘆醇比”灯扣⑺丨治 療組之大白鼠體重比控制組減少約6%。 綜上所述,藉由本發明可知,白黎蘆醇resveratr〇l確實能有效地降低血 糖;它同時對胰島素依賴型(IDDM)與胰島素非依賴型(NIDDM)糖尿病皆具 療效,並可延緩胰島素耐受性之產生且有控制體重之效果,因此確實可有效 改善現在使用之降血糖藥物所衍生之缺失,實為一種極佳之發明創作,且本 發明並未見於任何刊物上,故完全符合發明專利之法定要件,爰依法提出申 請專利’懇請鈞局賜准本發明專利。 歸納上述所說,本發明同時具有上述眾多效能與實用價值,並可有效提 升整體的經濟效益,因此本發明確實為一創意極佳的發明,且在相同技術領 域中未見相同或近似之產品公開使用,應已符合發明專利之要件,乃依法提 出申請,並請賜予本發明專利。 【圖式簡單說明】 第一圖係本發明中白黎蘆醇resveratr〇l對正常大白鼠(仰)之降血糖 作用示意圖。 第二圖係本發明中白黎蘆醇resveratrol對胰島素依賴型(IDDM)大白 鼠之降血糖作用示意圖。 第三圖係本發明中白黎蘆醇resveratrol對非胰島素依賴型(NIDDM) — 騰島素耐受型大白鼠之降血糖作用示意圖。 第四圖係本發明之白黎蘆醇resveratrol對非胰島素依賴型(NIDDM)— 1258010 肥胖型大白鼠之降血糖作用示意圖。 第五圖係本發明中比較口服白黎蘆醇resveratrol後對SD、IDDM、 NIDDM —胰島素耐受型與nidDM —肥胖型大白鼠之降血糖作用示意圖。 第六圖係本發明中比較NIDDM大白鼠中白黎産醇resveratrol治療組與 控制組之體重示意圖。 【主要元件符號說明】 無5, blood glucose detection method: First, the night before the experiment, SD mice, STZ or NIDDM diabetic mice were fasted. The next day, the rats were anesthetized with a pent〇 barbital 3 mg mg/kg intraperitoneally. After taking fasting blood glucose 〇·〇6 ml, the rats were given white resverat resveratr〇1 in the body of the mice. After 60, 90, and 120 minutes, blood was again drawn for centrifugation (13 rpm, 3 minutes) to separate serum and plasma. Take the upper serum 1 ul, add gluc〇se kit reaction reagent i 〇 ml, mix well, and react in a constant temperature water bath at 37 ° C for 10 minutes. The blood glucose level is then measured by the difference in absorbance using a blood glucose meter (Quick-Lab, Chemistry Analyzer). The principle of the Gluc〇se kit reagent is the oxidation of glucose to gluconic acid and hydrogen peroxide, which is oxidized by the pigment base of glucose to give a different shade of red. The blood glucose level is red expressed by the reaction of serum and gluc〇se kit; the color change can be perceived by the difference in absorbance, and the blood glucose value is calculated according to the standard product. (mg/dL) ° 6. Statistical analysis: The results obtained by the experiment are averaged Value ± standard error (mean ± SE) to indicate, and Student's t-test to assess the difference between the two, the comparison of the two groups using AN〇VA and Dunnett's Post-hoc test to assess the difference. p<〇 〇5, which is considered to be a significant difference. The experimental results of the present invention are as follows: 1. The hypoglycemic effect of resveratrol on normal rats (SD): Please refer to the first figure to give SD rats different doses of resvermrel resvemtr〇. 1 (〇〇5, 0·1, 0.5 mg/kg). After 90 minutes, it was found that resveratrol resveratr〇i with the increase of dose, 8 1258010 can obviously produce hypoglycemic effect on SD rats. The hypoglycemic effect of resveratrol 〇1 mg/kg was about 25.72±4·03%. At a dose of 0.5 mg/kg, resveratrol produced by resveratrol had the highest blood glucose lowering effect of about 35.53 ± 2.0%. 2, resveratrol resveratrol on insulin-dependent (IDDM) rats hypoglycemic effect: Please refer to the second figure, give IDDM rats oral doses of different doses of resveratrol resveratrol (0·05, 0· 1, 0·5 mg/kg). After 90 minutes, resveratr〇i resveratr〇i was found to significantly reduce blood glucose in IDDM mice with increasing dose. The hypoglycemic effect of resveratrol 0.1 mg/kg was about 21·00±5.84%. At a dose of 0.5 mg/kg, the effect of blood glucose lowering by resveratrol was the highest, about 1.89% of 23 soil. 3, resveratrol resveratrol for non-insulin-dependent (NIDDM) rats hypoglycemic effect: please refer to the third and fourth figures, given NIDDM rats oral doses of different doses of resveratrol resveratrol (0 ·05, 0.1, 0.5 mg/kg). After 90 minutes, it was found that resveratrol resveratrol can significantly reduce blood glucose in NIDDM mice with increasing dose. The hypoglycemic effect of the Tengdao vegetarian-type-NIDDM-fed white resveratrol 0.1 mg/kg can reach 28.3±1.66%. Moreover, at a dose of mg·5 mg/kg, the effect of resveratrol produced by resveratrol can be maximized, about 32.UL 62%. The hypoglycemic effect of obesity-NIDDM on white JI alcohol resveratr〇l 〇·ι mg/kg can reach 26.43 ±2.48% 〇4, compared with oral resveratrol resveratr〇l to normal, IDDM, Hypoglycemic effect of NIDDM rats: Please refer to the fifth figure, the hypoglycemic effect of oral resveratrol resveratrol 0.5 mg/kg after 90 minutes, in the presence of insulin, NIDDM-insulin and NIDDM- The percentage of hypoglycemic blood glucose produced by nicotinamide rats was 35.53±2.0%, 32·1±1·62%, 24.86±1·32% (N=8), which was produced in insulin-free IDDM rats. The percentage of blood glucose lowering was 23·00±1·89% (Ν=8). 1258010 5, resveratrol (10) veratrol delays the insulin resistance of SD rats. · Please refer to the sixth figure, when the normal SD rat insulin (〇.5lu/kg, Tid) is administered every day, the rats Insulin resistance (insuHn resistance) began to occur at 16.5 ± 1.5 days, while resveratrol resvemtrol (0.5 mg/kg, Bid) treatment group effectively delayed the development of insulin resistance, at 23 Animals develop insulin resistance at 5 days and 1.5 days. At the same time, we also found that resveratrol has a body weight reduction of about 6% compared with the control group of the lamp deduction (7) treatment group. In summary, by the present invention, resveratrol resveratr〇l can be effectively reduced. Blood sugar; it is effective against both insulin-dependent (IDDM) and insulin-independent (NIDDM) diabetes, and delays the development of insulin resistance and has a weight-control effect, so it can effectively improve the blood sugar lowering used now. The deficiency derived from the drug is an excellent invention creation, and the invention is not found in any publication, so it fully complies with the statutory requirements of the invention patent, and the patent application is filed according to law. In view of the above, the present invention has the above-mentioned numerous performances and practical values, and can effectively improve the overall economic benefit. Therefore, the present invention is indeed an innovative invention, and the same or similar products are not disclosed in the same technical field. Use, should have met the requirements of the invention patent, is to apply according to law, and please give the invention patent. [Simplified description] The first figure is a schematic diagram of the hypoglycemic effect of resveratrol 〇 res resveratr 〇l on normal rats (Yang) in the present invention. The second figure is the resveratrol resveratrol in the present invention for insulin-dependent (IDDM) rats. Schematic diagram of blood glucose action. The third figure is a hypoglycemic effect of resveratrol resveratrol on non-insulin-dependent (NIDDM)-Tengdaosu-tolerant rats in the present invention. The fourth figure is the resveratrol of the present invention. Schematic diagram of the hypoglycemic effect of resveratrol on non-insulin-dependent (NIDDM)-1258010 obese rats. The fifth figure is the comparison of oral resveratrol resveratrol to SD, IDDM, NIDDM-insulin-tolerant and nidDM. Schematic diagram of hypoglycemic effect of obese rats. Fig. 6 is a schematic diagram comparing the body weight of the resveratrol treatment group and the control group of NIDDM rats in the present invention. [Explanation of main component symbols]

1111

Claims (1)

1258010 拾、申請專利範圍: 9i〇- 1、一種證明白黎蘆醇具有降也糖藥效的方法,係準備一正常邪 大白鼠、胰島素依賴型IDDM及非胰島素依賴型NIDDM的糖尿病大白鼠並 分別給予口服一不同劑量的白黎蘆醇(resveratr〇1),其中該白黎蘆醇 (resveratrol)係溶於生理食鹽水後,以〇. 〇5、〇 !、〇 5 mg/ml之劑量, 再經由給藥管灌食到不同型的糖尿病大白鼠的胃部,在經過一適當時間後, 觀察該白黎蘆醇(resveratrol)隨著劑量的增加,對上述之大白鼠產生降 血糖的作用。 2、 如申請專利範圍第1項所述之一種證明白黎蘆醇具有降血糖藥 效的方法,其中,該正常SD大白係為購自國家動物中心的Sprage Dawley (SD)品系的雄性大白鼠,年齡在8週以上,體重約2〇〇-25〇g之間,飼養於 長庚醫院動物中心以空調維持室溫在25±lt的動物室,光照時間與黑暗時間 各為12小時,且讓老鼠自由的進食與飲水。 3、 如申請專利範圍第!項所述之一種證明白黎蘆醇具有降血糖藥 效的方法,其中,該胰島素依賴型麵的糖尿病老鼠係為將8週的幼雄 性大白鼠禁食72小時後,以腹腔注射pent〇barbital 3〇 _g將之麻醉, 自股靜脈注射,給予老鼠strept_tc)cin ( STZ,65嗯他),並於^小 時後,抽取靜脈血,利用一葡萄糖試劑(gluc〇se Kit),經由一自動分析儀 (Autoanalyser,Quik—Lab·,Ames)得知血糖值,當老鼠的血糖值大於侧 mg/dL ’並出現多吃、多喝、多尿症狀,即視為騰島素依賴型(ι_糖尿病 老鼠。 4、 如申請專利範圍第1項所述之一種證明白黎蘆醇具有降血糖藥 效的方法’其中,該非胰島素依賴型N_的糖尿病老鼠係為:胰島素耐 受性型(NI_-insulin):利用SD系的雄性大白鼠週齡8週大以皮下 注射給予長效型的胰島素’每天投予三次,每次劑量〇·5 ,如此持續 12 1258010 少兩個星期,·肥胖型(NIDDM_nicotinamide) ·· SD雄性大白鼠禁食48小時 後’以 pentobarbital 30 mg/kg 麻醉後’先腹腔注射 nicotinamide 100 mg/kg 後再由股靜脈注射streptozotocin( STZ,65 mg/kg),並於四周後測其血糖 值’當該血糖值大約等於200 mg/dL,並產生肥胖現象。 5、一種白黎蘆醇藥劑,其特徵在於: 該藥劑至少包含有〇· 〇5 mg/mi至〇· 5 mg/ml劑量之白黎董醇 (resveratrol),可達到促進血糖代謝及延遲胰島素耐受性產生之效果。1258010 Pick up, apply for patent scope: 9i〇-1, a method to prove that resveratrol has the effect of reducing sugar, prepare a normal white rat, insulin-dependent IDDM and non-insulin-dependent NIDDM diabetic rats Each dose of resveratrol (resveratr〇1) was administered orally, wherein the resveratrol was dissolved in physiological saline, and the dose was 〇5, 〇!, 〇5 mg/ml. And then feeding through the drug tube to the stomach of different types of diabetic rats. After a suitable period of time, the resveratrol is observed to increase blood glucose in the above-mentioned rats as the dose increases. effect. 2. A method for demonstrating that resveratrol has hypoglycemic effect as described in claim 1, wherein the normal SD white line is a male rat of the Sprage Dawley (SD) strain purchased from the National Animal Center. , aged over 8 weeks, weighing between 2〇〇-25〇g, kept in the animal center of Chang Gung Memorial Hospital with air conditioning to maintain room temperature at 25±lt in the animal room, the light time and dark time are each 12 hours, and let Rats are free to eat and drink. 3. If you apply for a patent scope! One of the methods described in claim that resveratrol has a hypoglycemic effect, wherein the insulin-dependent profiled diabetic mouse is intraperitoneally injected with pent〇barbital after fasting for 8 hours in 8 weeks of juvenile male rats. 3〇_g anesthetize, from the femoral vein, give the mouse strept_tc)cin (STZ, 65 hm he), and after ^ hours, take venous blood, using a glucose reagent (gluc〇se Kit), via an automatic The analyzer (Autoanalyser, Quik-Lab·, Ames) knows the blood sugar level. When the blood sugar level of the mouse is greater than the side mg/dL 'and the symptoms of eating more, drinking more, and more urine, it is considered to be dependent on the island. _ Diabetic rats 4. A method for demonstrating that resveratrol has hypoglycemic effect as described in claim 1 wherein the non-insulin dependent N_ diabetic mouse is insulin resistant ( NI_-insulin): male rats of the SD line were given a subcutaneous injection of long-acting insulin at a peripheral age of 8 weeks, three times a day, each dose 〇·5, so lasting 12 1258010 less than two weeks, · obesity Type (NIDDM_nicot Inamide) · SD male rats were fasted for 48 hours after 'anesthesia with pentobarbital 30 mg/kg'. After intraperitoneal injection of nicotinamide 100 mg/kg, streptozotocin (STZ, 65 mg/kg) was injected from the femoral vein and surrounded by four weeks. After the blood glucose value is measured 'when the blood sugar value is approximately equal to 200 mg / dL, and obesity occurs. 5. A resveratrol agent, characterized in that the agent contains at least 〇 · 〇 5 mg / mi to 〇 · Resveratrol at a dose of 5 mg/ml can achieve the effects of promoting blood sugar metabolism and delaying insulin resistance. 1313
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