TW202017577A - 冬蟲夏草的萃取物用於提升端粒酶反轉錄酶基因及端粒酶rna組分基因的表現量及抗老化之用途 - Google Patents
冬蟲夏草的萃取物用於提升端粒酶反轉錄酶基因及端粒酶rna組分基因的表現量及抗老化之用途 Download PDFInfo
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Abstract
本發明提供一種冬蟲夏草的萃取物用於提升端粒酶反轉錄酶基因及端粒酶RNA組分基因的表現量,及抗老化的用途。
Description
本發明是有關於一種冬蟲夏草(Hirsutella sinensis)的萃取物用於提升端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因及端粒酶RNA組分(telomerase RNA component,TERC)基因的表現量,及抗老化的用途。
自古人類總夢想著追求青春永駐、長生不老之道,近年來隨著醫學及生物科技的發展與進步,不但可藉由前端的醫學技術來對抗疾病,許多訴求能抗老的產品也陸續地研發問世,近年來抗老化的風潮慢慢地向全球蔓延開來,以日本為例,高達七成以上民眾具有抗老化意識,而台灣民眾對於抗老化的關心也持續升高,此股抗老化趨勢將帶動抗老化相關產品市場銷售,預期全球市場也將持續擴張。
端粒酶(telomerase)是一種由RNA和蛋白質組成的核糖核蛋白複合體,與端粒的調控機理密切相關。端粒酶可延長端粒修復能力,減少端粒因細胞分裂而損耗,讓細胞分裂複製的次數增加。端粒酶活性(telomerase activity)亦與細胞增生及存活有關,細胞會持續失去端粒酶活性,且DNA修補過程中,也會失去端粒DNA,因此端粒酶活性與細胞老化有關聯性。
目前常見用來延緩老化問題的方式大多為利用塗抹於皮膚表面的化妝品、保養品,或口服宣稱具有抗老化效用的健康食品。然而,習知的化妝品、保養品及健康食品大多由化學成分所製成,長期使用不但對人體健康有害無益,且這些產品往往價格昂貴,並非為一般使用者所能負擔。
為了解決上述問題,本領域的技術人員亟需研發出具有提升端粒酶活性及抗老化之功效的新穎醫藥組成物或食品產品以造福有此需求的廣大族群。
有鑑於此,本發明之目的為提供一種冬蟲夏草(Hirsutella sinensis)的萃取物用於製備一提升端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因及端粒酶RNA組分(telomerase RNA component,TERC)基因的表現量之組成物的用途。
本發明之另一目的為提供一種冬蟲夏草的萃取物用於製備一抗老化之組成物的用途。
在本發明的一實施例中,冬蟲夏草的萃取物是以水、醇類、含水醇類或其組合作為一萃取溶劑對冬蟲夏草進行萃取而製得。
在本發明的一實施例中,冬蟲夏草與萃取溶劑的體積比例介於1~5:5~20。
在本發明的一實施例中,萃取的溫度介於60℃至90℃。
在本發明的一實施例中,組成物是一醫藥組成物或一食品產品。
綜上所述,本發明冬蟲夏草的萃取物之功效在於:可藉由提升端粒酶反轉錄酶基因及端粒酶RNA組分基因的表現量來提升端粒酶活性,進而達到抗老化的功效。
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是本發明冬蟲夏草的萃取物在調控端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因表現上的功效之數據圖,其中“*”表示與對照組比較,p<0.05。
圖2是本發明冬蟲夏草的萃取物在提升端粒酶RNA組分(telomerase RNA component,TERC)基因表現上的功效之數據圖。
本文中所使用數值為近似值,所有實驗數據皆表示在20%的範圍內,較佳為在10%的範圍內,最佳為在5%的範圍內。
依據本發明,冬蟲夏草(Hirsutella sinensis)別名為蟲草、冬蟲草或中華被毛孢,為麥角菌科(Clavicipitaceae)被毛孢黴屬(Hirsutella)的真菌生物,亦為中國歷史中傳統的名貴中藥材。冬蟲夏草主要產於中國大陸青海、西藏、四川、雲南、甘肅和貴州等省及自治區的高寒地帶和雪山草原。冬蟲夏草已知可被用來抗腫瘤、降血糖、降血脂、改善慢性腎臟病及保護肝臟。
如本文中所使用的,用語「抗老化(anti-aging)」意指預防、減緩人類皮膚外觀之老化現象,例如:皺紋的產生及失去彈性等。評量實現此目的之程度將根據熟悉此項技藝者已知之諸多因素來決定,諸如消費者的全身狀態、年齡、性別等。
依據本發明,醫藥組成物可利用熟習此技藝者所詳知的技術而被製造成一適合於非經腸道地(parenterally)或口服地(orally)投藥的劑型,這包括,但不限於,注射品(injection)[例如,無菌的水性溶液(sterile aqueous solution)或分散液(dispersion)]、無菌的粉末(sterile powder)、錠劑(tablet)、片劑(troche)、丸劑(pill)、膠囊(capsule)以及類似之物。
依據本發明,醫藥組成物可以一選自於下列所構成的群組中的非經腸道途徑來投藥:腹膜內注射(intraperitoneal injection)、皮下注射(subcutaneous injection)、肌肉內注射(intramuscular injection)、靜脈內注射(intravenous injection)、舌下投藥(sublingual administration)以及穿皮投藥(transdermal administration)。
依據本發明,醫藥組成物可進一步包含有一被廣泛地使用於藥物製造技術之醫藥上可接受的載劑(pharmaceutically acceptable carrier)。例如,該醫藥上可接受的載劑可包含一或多種選自於下列的試劑:溶劑(solvent)、緩衝液(buffer)、乳化劑(emulsifier)、懸浮劑(suspending agent)、分解劑(decomposer)、崩解劑(disintegrating agent)、分散劑(dispersing agent)、黏結劑(binding agent)、賦形劑(excipient)、安定劑(stabilizing agent)、螯合劑(chelating agent)、稀釋劑(diluent)、膠凝劑(gelling agent)、防腐劑(preservative)、潤濕劑(wetting agent)、潤滑劑(lubricant)、吸收延遲劑(absorption delaying agent)、脂質體(liposome)以及類似之物。有關這些試劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。
依據本發明,該醫藥上可接受的載劑包含有一選自於由下列所構成之群組中的溶劑:水、生理鹽水(normal saline)、磷酸鹽緩衝生理鹽水(phosphate buffered saline,PBS)、含有醇的水性溶液(aqueous solution containing alcohol)以及它們的組合。
依據本發明,組成物可被當作食品添加物(food additive),藉由習知方法於原料製備時添加,或是於食品的製作過程中添加,而與任一種可食性材料配製成供人類與非人類動物攝食的食品產品。
依據本發明,食品產品的種類包括但不限於:飲料(beverages)、發酵食品(fermented foods)、烘培產品(bakery products)、健康食品(health foods)以及膳食補充品(dietary supplements)。
首先,將冬蟲夏草進行均質,接而以水、醇類、含水醇類或其組合作為萃取溶劑對均質後的冬蟲夏草進行萃取1~3小時,其中萃取的溫度介於60℃至90℃,萃取溶劑與冬蟲夏草的體積比介於5~20:1~5。接著,將所得到的產物進行離心,接而取離心後的上清液並將之過濾而得到一濾液。之後,將濾液於45℃至70℃進行減壓濃縮而得到本發明冬蟲夏草的萃取物。
首先,以添加10% FBS、0.1mM非必需胺基酸、1.5g/L碳酸氫鈉、1mM丙酮酸鈉之MEM培養基培養人類纖維母細胞CCD966SK於6-孔盤,2mL培養基的細胞濃度為1.5×105細胞/孔,接而於37℃進行培養。
之後,將經培養的細胞分成3組,其中包括1個對照組及2個實驗組(亦即實驗組1及實驗組2)。將0.0625mg/mL濃度的冬蟲夏草的水萃取物添加至實驗組1的細胞中,及將0.0625mg/mL濃度的冬蟲夏草的乙醇萃取物添加至實驗組2的細胞中,至於對照組的細胞則不做任何處理。進行培養24小時後,收取各組細胞培養物並拿來進行基因表現分析。
在本實施例中,用來分析與端粒酶活性相關的基因包括端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因及端粒酶RNA組分(telomerase RNA component,TERC)基因。
以RNA萃取套組(RNA extraction kit)(Geneaid)對上面所得到的各組細胞培養物進行RNA的萃取。之後,以SuperScript® III反轉錄酶(SuperScript® III Reverse Transcriptase)(Invitrogen)將萃取出的RNA反轉錄為cDNA。接著,以cDNA作為模版(template),並且使用擴增標的基因的引子對包括TERT、TERC及GAPDH(作為內部對照組),它們的核苷酸序列顯示於下面表1,利用KAPA CYBR FAST qPCR套組(2x)(KAPA Biosystems)來進行定量即時聚合酶鏈反應(quantitative real-time polymerase chain reaction,以下簡稱定量即時PCR),俾以對標的基因進行擴增及定量。PCR產物的熔化曲線(melting curve)是在定量即時PCR反應期間進行確認。
標的基因的相對表現量是推導自方程式2-△△Ct,並利用GAPDH基因(作為內部對照組)及基準基因(reference gene)的循環閾值[cycle threshold(C t)value]及藉由標準差(standard deviation,STDEV)來計算相對倍數變化,其中△Ct=Ct目標基因/基準基因-CtGAPDH,△△Ct=△Ct目標基因-△Ct基準基因,倍數變化=2-△△Ct 平均值。以對照組的標的基因表現量作為1的比較基準。各組之間的統計學顯著差異是藉由單尾史徒登氏t-檢定(single tailed Student’s t-test)來決定。本實施例的結果顯示於圖1及圖2。
圖1是本發明冬蟲夏草的萃取物在調控端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因表現上的功效之數據圖。圖2是本發明冬蟲夏草的萃取物在提升端粒酶RNA組分(telomerase RNA component,TERC)基因表現上的功效之數據圖。由圖1可見,與對照組相較之下,實驗組1測得的TERT基因相對表現量有顯著提升。由圖2可見,與對照組相較之下,實驗組1及實驗組2測得的TERC基因相對表現量皆有顯著提升。本實施例的結果顯示,本
發明冬蟲夏草的萃取物可藉由調控與端粒酶活性相關的基因表現來達到抗老化的功效。
綜上所述,本發明冬蟲夏草的萃取物可藉由提升端粒酶反轉錄酶基因及端粒酶RNA組分基因的表現量來提升端粒酶活性,進而達到抗老化的功效。
以上所述僅為舉例性,而非為限制性者。任何未脫離本發明之精神與範疇,而對其進行之等效修改或變更,均應包含於後附之申請專利範圍中。
<110> 大江生醫股份有限公司
<120> 冬蟲夏草的萃取物用於提升端粒酶反轉錄酶基因及端粒酶RNA組分基因的表現量及抗老化之用途
<130> 107B0472-I1
<160> 6
<170> PatentIn version 3.5
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Claims (10)
- 一種冬蟲夏草(Hirsutella sinensis)的萃取物用於製備一提升端粒酶反轉錄酶(telomerase reverse transcriptase,TERT)基因及端粒酶RNA組分(telomerase RNA component,TERC)基因的表現量之組成物的用途。
- 如申請專利範圍第1項所述的用途,其中該冬蟲夏草的萃取物是以水、醇類、含水醇類或其組合作為一萃取溶劑對該冬蟲夏草進行萃取而製得。
- 如申請專利範圍第2項所述的用途,其中該冬蟲夏草與該萃取溶劑的體積比例介於1~5:5~20。
- 如申請專利範圍第2項所述的用途,其中該萃取的溫度介於60℃至90℃。
- 如申請專利範圍第1項所述的用途,其中該組成物是一醫藥組成物或一食品產品。
- 一種冬蟲夏草(Hirsutella sinensis)的萃取物用於製備一抗老化之組成物的用途。
- 如申請專利範圍第6項所述的用途,其中該冬蟲夏草的萃取物是以水、醇類、含水醇類或其組合作為一萃取溶劑對該冬蟲夏草進行萃取而製得。
- 如申請專利範圍第7項所述的用途,其中該冬蟲夏草與該萃取溶劑的體積比例介於1~5:5~20。
- 如申請專利範圍第7項所述的用途,其中該萃取的溫度介於60℃至90℃。
- 如申請專利範圍第6項所述的用途,其中該組成物是一醫藥組成物或一食品產品。
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CN201910466554.3A CN111166773A (zh) | 2018-11-13 | 2019-05-30 | 冬虫夏草的萃取物用于提升基因表现量及抗老化的用途 |
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