TWI740030B - 包含植物萃取物的組成物及其於促進glut4基因表現之用途 - Google Patents
包含植物萃取物的組成物及其於促進glut4基因表現之用途 Download PDFInfo
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- TWI740030B TWI740030B TW107111697A TW107111697A TWI740030B TW I740030 B TWI740030 B TW I740030B TW 107111697 A TW107111697 A TW 107111697A TW 107111697 A TW107111697 A TW 107111697A TW I740030 B TWI740030 B TW I740030B
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- composition
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- green tea
- glut4
- gene expression
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Abstract
本發明係關於一種包含植物萃取物的組成物,提供了兩種植物萃取物所組合的組成物,相較於單一成分的萃取物,本發明之萃取物組合更能夠有效提升GLUT4基因的表現。構成所述組成物之萃取物可包括:柑橘萃取物與綠茶萃取物。
Description
本發明係關於一種包含植物萃取物之組成物及其用途,尤其是關於一種可促進GLUT4基因表現而具有調節血醣功效之組成物及其用途。
所謂血醣,係指血液中的葡萄糖,其來自於小腸的消化,並可進一步藉著血液將其輸送到身體中的各個細胞,作為細胞主要的能量來源。於人體中,血醣的濃度是被嚴格調節與控制的,一般濃度約為900mg/L(即5mmol/L或90mg/dl)左右,正常範圍在4-6mmol/L之間,使維持在一恆定的濃度範圍中,但隨著飲食與作息,會有較大的波動產生,一般進食後一到兩個小時血醣濃度會升高,而在早晨時則會降到最低。血醣濃度失調會導致多種疾病,特別是持續血醣濃度過高時就可能引發糖尿病。
糖尿病可分為胰島素無法產生或產生不足的第一型糖尿病,以及細胞對於胰島素的反應不正常或沒有反應的第二型糖尿病。不論是哪一型糖尿病,如果不進行治療,將引發諸多併發症,例如:糖尿病酮酸血症、高滲透壓高血醣非酮酸性昏迷,以及心血管疾病、慢性腎臟病、視網膜病變、糖尿病足、中風等。據統計,全世界糖尿病患人口於2016年時約有4.22億,眾多患者苦於併發症,生活品質下降,生命風險更是提升。因此,關於血醣的調控一直是科學家們汲汲營營努力所欲解決的目標。
葡萄糖載體蛋白4(glucose transporter 4,GLUT4),與葡萄糖的運送相關,主要存在於肌肉及軟骨組織中,該蛋白在細胞質中時就可結合裝載有胰島素,一旦血液中葡萄糖濃度升高,就會刺激葡萄糖載體蛋白4位移至細胞膜上,釋放出胰島素以迅速調節血醣,同時也可結合葡萄糖,將其攜帶回細胞中。因此,葡萄糖載體蛋白4的基因表現若能增加,將可產生更多的葡萄糖載體蛋白4,而提升對胰島素的敏感度,達到調整或降低血醣之目的。
在國人日常所常食用或飲用之植物或保健成分中,例如:紅酒、柑橘、藍莓、綠茶等,並未知悉其單獨或組合是否具有促進GLUT4基因表現之效用,若又能從複方的組成物中,找尋出可大幅提升GLUT4基因表現之組份組合,對於相關糖尿病患者而言將是一大福音。
本發明目的之一在於提供一種包含植物萃取物的組成物,藉由其本身,或製備成醫藥組成物或食品組成物,在結合兩種植物萃取物作用之情況下,大幅促進GLUT4基因的表現,達到使個體在血醣濃度上能獲得適度調節或降低的目的。
在本發明的一實施例中,該組成物係包含柑橘萃取物與綠茶萃取物。
在本發明的一實施例中,其中該柑橘萃取物相對於綠茶萃取物之比例為0.8~1.2:1.2~0.8,較佳為1:1。在一態樣中,該柑橘萃取物與該綠茶萃取物之濃度可分別為0.03125~0.25mg/ml。
在本發明的另一實施例中,提供一種具促進GLUT4基因表現的醫藥組成物,可將前述組成分製備成一醫藥組成物。
在本發明的前述實施例中,該醫藥組成物可包括前述的組成物,以及一藥學上可接受之載劑。
在本發明的前述實施例中,該醫藥組成物之劑型可為溶液、膠囊或錠劑。
本發明之再一實施例中,提供一種具促進GLUT4基因表現的食品組成物。
在本發明的前述實施例中,該食品組成物可包括前述的組成物,以及一食品成分,而該食品成分可為一般食品、保健食品、膳食補充品或飲品之成分。
本發明的另一實施例中,提供一種包含植物萃取物之組成物的用途,利用該組成物來促進GLUT4基因表現,而該組成物可為前述的組成物。
在本發明的前述實施例中,該包含植物萃取物之組成物可進一步用以製造促進GLUT4基因表現之醫藥組成物,或用以製造促進GLUT4基因表現之食品、保健食品、膳食補充品或飲品。
以下將進一步說明本發明的實施方式,下述所列舉的實施例係用以闡明本發明,並非用以限定本發明之範圍,任何熟習此技藝者,在不脫離本發明之精神和範圍內,當可做些許更動與潤飾,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。
圖1係本發明實施例中不同植物萃取物或組合對於GLUT4基因表現影響之結果比較圖。
將綠茶(green tea)洗淨及乾燥,以粉碎機將綠茶粗碎,之後以水為溶劑,以該溶劑與綠茶粗碎物依液固比5~20:1~5的比例混合後,於50℃~100℃下對綠茶粗碎物進行萃取0.5~3小時。萃取溫度較佳為75℃~95℃。
經上述萃取步驟所得綠茶萃取物冷卻至室溫後,以400目(mesh)濾網過濾以移除殘餘固體物。過濾後的綠茶萃取物可進一步在45℃~70℃下進行減壓濃縮而獲得一濃縮產物。
在本實施例中,較佳地,水與綠茶粗碎物可依液固比5:1的比例混合後,於75℃下對綠茶粗碎物進行粹取3小時,經前述萃取步驟所得綠茶萃取物冷卻至室溫後,以400目(mesh)濾網過濾以移除殘餘固體物。過濾後的綠茶萃取物可進一步在70℃下進行減壓濃縮而獲得一濃縮產物。
本發明實施例之柑橘萃取物係對柑橘(Citrus reticulata)果實進行萃取而獲得,該萃取物可購自羅頓貿易有限公司。紅酒萃取物係對紅葡萄酒進行萃取而獲得,該萃取物可購自上海博優堂生物技術有限公司。青咖啡豆萃取物係對未經烘焙的咖啡屬植物(Coffea spp.)種子進行萃取而獲得,該萃取物可購自ARJUNA NATURAL EXTRACTS有限公司(印度)。藍莓萃取物係對北美藍莓(Vaccinium Cyanococcus)果實進行萃取而獲得,該萃取物可購自元弘草本科研有限公司。將該些萃取物以水配製成適當濃度後備用。
準備HepG2細胞(購自ATCC HB8065),培養於細胞培養液[含有1%盤尼西林/鏈黴素(Penicillin/streptomycin)、10%胎牛血清的Dulbecco's modified Eagle's培養液(Gibco)]中。於6孔盤每孔中加入2ml細胞培養液,使每孔具有1.5 x 105個HepG2細胞。
之後將樣本分為二組,其中,A組為空白控制組,B組則為試驗組。試驗組B中分別依照表1所列之植物萃取物、組份種類,分別以0.03125mg/ml至0.5mg/ml的濃度,以個別萃取物,或兩萃取物組合以1:1的比例加入由前述實施例1或2中所製備的萃取物後,於37℃下反應6與24小時,之後進行GLUT4基因表現量狀況之分析。表中所示濃度,係個別萃取物、或組合萃取物加入培養基後中各組份在培養基中的濃度。
將上述各組細胞回收,以RNA萃取套組(Genemark)萃取其RNA,並以反轉錄酶(SuperScript III Reverse Transcripatase,Invitrogen)將該些RNA(2000ng)反轉錄為cDNA。之後利用即時聚合酶鏈鎖反應系統(ABI Step One Plus Real-Time PCR system),利用表2所列引子對進行qPCR(KAPA CYBR FAST qPCR Kits,KAPA Biosystems),以定量GLUT4基因的表現[以ACTB(β-肌動蛋白,beta-actin)基因的表現做為內部控制組]。即時聚合酶鏈鎖反應過程中,分析其解離曲線(melting curve),並採用SCORE法,以前述ACTB基因的循環閥值(cycle threshold,Ct)做為內部控制組,以空白控制組為對照基因,進行基因表現相對定量分析。以SCORE法分析時,目標基因(GLUT4)的mRNA相對量係推導自方程式2-△Ct,其中△Ct=Ct目標基因-CtACTB,接著計算GLUT4基因之表現量在各組與空白控制組中的差值,並以此差值之加總作為血醣基因指數。基因表現所對應的血醣基因指數,其結果如表1與圖1所示,圖1為表1圖示化的柱狀圖。指數數值愈高,代表對於GLUT4基因表現的促進效果愈好。
由表1與圖1之結果可知,單獨柑橘萃取物(0.25mg/ml)對於GLUT4基因表現的促進效果之指數為1.10,單獨綠茶萃取物(0.25mg/ml)的促進效果之指數為0,但將二者組合作用後,其促進效果之指數可高達2.84,相對於原來柑橘之效果,大幅提高至258%。相較之下,若將柑橘萃取物與藍莓萃取物組合後加以作用,反而使促進效果之指數降為0。因此,關於萃取物組份間之組合效果,非必然係相乘效果,於本發明之實施例中,係以柑橘萃取物結合綠茶萃取物所獲得之促進效果為較佳。
藉由上述試驗可知,本發明實施例之具促進GLUT4基因表現的組成物,在結合柑橘萃取物與綠茶萃取物兩種萃取物的情況下,可產生超乎預期的相乘促進效果,使GLUT4基因的表現顯著增加。因此,可利用該些組成物製備相關的醫藥或食品組成物,使施用者或食用者在血醣濃度獲得調解,而可藉以改善血醣過高的情形。
此外,本發明實施例之具促進GLUT4基因表現的組成物,亦可進一步添加於食品、保健食品、膳食補充品或飲品中。將本發明實施例之具促進GLUT4基因表現的組成物製備為醫藥組成物時,該醫藥組成物亦可進一步加入所屬技術領域所熟知之載劑或其他輔劑。醫藥組成物之劑型,可為但不限於溶液、膠囊、或錠劑。
<110> 大江生醫股份有限公司
<120> 包含植物萃取物的組成物及其於促進GLUT4基因表現之用途
<130> 107B0133-I1
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 用於PCR的正向引子
<210> 2
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 用於PCR的反向引子
Claims (5)
- 一種由柑橘萃取物與綠茶萃取物所組成之組成物用於製備一促進GLUT4基因表現之組成物的用途,其中該柑橘萃取物相對於該綠茶萃取物之比例為0.8~1.2:1.2~0.8。
- 如申請專利範圍第1項所述之用途,其中該由柑橘萃取物與綠茶萃取物所組成之組成物係為醫藥組成物。
- 如申請專利範圍第1項所述之用途,其中該由柑橘萃取物與綠茶萃取物所組成之組成物係為食品、保健食品、膳食補充品或飲品。
- 如申請專利範圍第1項所述之用途,其中該柑橘萃取物與該綠茶萃取物之濃度分別為0.03125~0.25mg/ml。
- 如申請專利範圍第1項所述之用途,其中該柑橘萃取物相對於該綠茶萃取物之比例為1:1。
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