CN110840932A - 波罗蜜白芯的萃取物用于调控基因表现量及改善停经症候群的用途 - Google Patents
波罗蜜白芯的萃取物用于调控基因表现量及改善停经症候群的用途 Download PDFInfo
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Abstract
本发明涉及植物提取物领域,尤其是关于一种波罗蜜白芯的萃取物用于调控基因表现量及改善停经症候群的用途。本发明提供的波罗蜜白芯的萃取物可用于调控CFOS基因、CSF1R基因、RANK基因、SYK基因、TRAP基因、APA1基因、ABCA1基因、IL‑6基因、VCAM1基因、CASP8基因、DDC基因、及ASMTL蛋白基因的表现量,及改善停经症候群。
Description
技术领域
本发明涉及植物提取物领域,尤其是关于一种波罗蜜(Artocarpusheterophyllus)白芯的萃取物用于调控基因表现量及改善停经症候群的用途。
背景技术
停经期(menopause)与女性体内循环的性激素(circulating sexual hormones)的快速减少有关联,并且意指女性的卵巢功能(ovarian function)逐渐衰退,最终导致月经终止(cessation of menstruation)的过渡期,而停经后期(postmenopause)则意指女性的卵巢功能在完全丧失之后的时期。停经期或停经后期的女性通常会出现停经症候群(menopausal syndrome),而停经症候群的病症主要包括:热潮红(hot flushes)、骨质疏松症(osteoporosis)、心血管疾病(cardiovascular disease)、失眠(insomnia)、皮肤老化(skin aging)、焦虑(anxiety)以及忧郁(depression)。
目前在停经症候群的治疗上大多是使用激素替代疗法(hormone replacementtreatment,HRT),藉由投药含有一或多种性激素的医药品给停经期或停经后期的女性来补充她们体内不足的性激素。然而,性激素的长期投药或过量使用可能会导致乳癌,并产生副作用。因此,若能开发出天然不具副作用,且可改善停经症候群的医药品或食品产品,将造福有此需求的广大族群并对本领域的技术带来相当大的突破。
发明内容
有鉴于此,本发明的目的为提供一种波罗蜜(Artocarpus heterophyllus)白芯的萃取物用于制备一调控CFOS基因、群落刺激因子1受体(colony stimulating factor 1receptor,CSF1R)基因、RANK基因、脾脏酪胺酸激酶(spleen tyrosinase kinase,SYK)基因、抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)基因、脂蛋白元A-1(apolipoprotein A-1,APA1)基因、ATP结合匣子族A成员1(ATP binding cassettesubfamily A member 1,ABCA1)基因、介白素6(interleukin-6,IL-6)基因、血管细胞黏附分子1(vascular cell adhesion molecule 1,VCAM1)基因、凋亡蛋白酶8(caspase 8,CASP8)基因、DOPA脱羧酶(DOPA decarboxylase,DDC)基因、及乙酰复合胺O-甲基转移酶类蛋白(Acetylserotonin O-methyltransferase-like protein,ASMTL protein)基因的表现量的医药品或食品产品的用途。
本发明的另一目的为提供一种波罗蜜白芯的萃取物用于制备一改善停经症候群的医药品或食品产品的用途。
在本发明的一实施例中,波罗蜜白芯的萃取物是以水或醇类作为萃取溶剂对波罗蜜白芯进行萃取而制得。
在本发明的一实施例中,萃取溶剂与波罗蜜白芯的液固比介于2~10∶1~5。
在本发明的一实施例中,萃取的温度介于50℃至100℃。
在本发明的一实施例中,萃取的时间介于0.5至3小时。
在本发明的一实施例中,医药品包含一医药上可接受的载剂。
在本发明的一实施例中,停经症候群包括骨质疏松症、心血管疾病、热潮红,及失眠。
综上所述,本发明波罗蜜白芯的萃取物的功效在于:可调控CFOS基因、CSF1R基因、RANK基因、SYK基因、TRAP基因、APA1基因、ABCA1基因、IL-6基因、VCAM1基因、CASP8基因、DDC基因、及ASMTL蛋白基因的表现量,并藉由抑制蚀骨细胞(osteoclast)分化与骨吸收、促进高密度脂蛋白(high density lipoprotein,HDL)生成与胆固醇代谢、减少内皮细胞发炎反应与细胞凋亡,及提升血清素(serotonin)与褪黑激素(melatonin)生成,达到改善停经症候群(包括骨质疏松症、心血管疾病、热潮红,及失眠)的功效。
以下将进一步说明本发明的实施方式,下述所列举的实施例是用以阐明本发明,并非用以限定本发明的范围,任何熟习此技艺者,在不脱离本发明的精神和范围内,当可做些许更动与润饰,因此本发明的保护范围当视后附的申请专利范围所界定者为准。
附图说明
图1是本发明波罗蜜白芯的萃取物在抑制蚀骨细胞分化上的功效的细胞染色图;
图2是本发明波罗蜜白芯的萃取物在调控与蚀骨细胞分化相关的基因(包括CFOS基因、CSF1R基因及RANK基因)表现上的功效的数据图,其中“***”表示与对照组2比较,p<0.001。;
图3是本发明波罗蜜白芯的萃取物在调控与蚀骨细胞骨吸收相关的基因(包括SYK基因及TRAP基因)表现上的功效的数据图,其中“***”表示与对照组2比较,p<0.001;
图4是本发明波罗蜜白芯的萃取物在调控与HDL生成及胆固醇代谢相关的基因(包括APA1基因及ABCA1基因)表现上的功效的数据图,其中“*”表示与对照组比较,p<0.05;“***”表示与对照组比较,p<0.001;
图5是本发明波罗蜜白芯的萃取物在调控与内皮细胞发炎反应与细胞凋亡相关的基因(包括IL-6基因、VCAM1基因及CASP8基因)表现上的功效的数据图,其中“*”表示与对照组比较,p<0.05;“***”表示与对照组比较,p<0.001;
图6是本发明波罗蜜白芯的萃取物在调控与色胺酸代谢相关的基因(包括DDC基因及ASMTL基因)表现上的功效的数据图,其中“***”表示与对照组比较,p<0.001。
具体实施方式
定义
本文中所使用数值为近似值,所有实验数据皆表示在20%的范围内,较佳为在10%的范围内,最佳为在5%的范围内。
如本文中所使用的,用语「停经期(menopause)」意指处于卵巢功能(ovarianfunction)开始衰退与月经终止(cessation of menstruation)之间的时期。
如本文中所使用的,用语「停经症候群(menopausal syndrome)」意指发生于停经期或停经后期的女性个体中的症状,这包括,但不限于:(1)生理症状(physiologicalsymptom),诸如热潮红(hot flushes)、***干燥(vaginal dryness)、疲劳(fatigue)、头痛(headaches)、心血管疾病(cardiovascular disease)、失眠(insomnia)、体重增加(weightgain)、皮肤老化(skin aging)以及骨质疏松症(osteoporosis);(2)心理症状(psychological symptom),诸如焦虑(anxiety)以及忧郁(depression);以及(3)认知缺陷(cognitive deficit),诸如精神错乱(mental confusion)、记忆能力(memory ability)下降、学习能力(learning ability)下降、辨识能力(recognition ability)下降以及无法应对(inability to cope)。
依据本发明的一实施例,波罗蜜(Artocarpus heterophyllus)别名为菠萝蜜、木波罗、树波罗或密冬瓜,为桑科(Moraceae)、波罗蜜属(Artocarpus)的常绿乔木。波罗蜜的果肉是水果,气味香甜,常用作烹调材料;种子也可食用,但生食可能会引起中毒。
依据本发明,医药品可利用熟习此艺者所详知的技术而被制造成一适合于非经肠地道(parenterally)或口服地(orally)投药的剂型,这包括,但不限于,注射品(injection)[例如,无菌的水性溶液(sterile aqueous solution)或分散液(dispersion)]、无菌的粉末(sterile powder)、锭剂(tablet)、片剂(troche)、丸剂(pill)、胶囊(capsule)以及类似物。
依据本发明,医药品可以一选自于下列所构成的群组中的非经肠道途径来投药:腹膜内注射(intraperitoneal injection)、皮下注射(subcutaneous injection)、肌肉内注射(intramuscular injection)、静脉内注射(intravenous injection)、舌下投药(sublingual administration)以及穿皮投药(transdermal administration)。
依据本发明,医药品可进一步包含有一被广泛地使用于药物制造技术的医药上可接受的载剂(pharmaceutically acceptable carrier)。例如,该医药上可接受的载剂可包含一或多种选自于下列的试剂:溶剂(solvent)、缓冲液(buffer)、乳化剂(emulsifier)、悬浮剂(suspending agent)、分解剂(decomposer)、崩解剂(disintegrating agent)、分散剂(dispersing agent)、黏结剂(binding agent)、赋形剂(excipient)、安定剂(stabilizingagent)、螯合剂(chelating agent)、稀释剂(diluent)、胶凝剂(gelling agent)、防腐剂(preservative)、润湿剂(wetting agent)、润滑剂(lubricant)、吸收延迟剂(absorptiondelaying agent)、脂质体(liposome)以及类似物。有关这些试剂的选用与数量是落在熟习此项技术的人士的专业素养与例行技术范畴内。
依据本发明,该医药上可接受的载剂包含有一选自于由下列所构成的群组中的溶剂:水、生理盐水(normal saline)、磷酸盐缓冲生理盐水(phosphate buffered saline,PBS)、含有醇的水性溶液(aqueous solution containing alcohol)以及它们的组合。
依据本发明,组成物可被当作食品添加物(food additive),藉由习知方法于原料制备时添加,或是于食品的制作过程中添加,而与任一种可食性材料配制成供人类与非人类动物摄食的食品产品。
依据本发明,食品产品的种类包括但不限于:饮料(beverages)、发酵食品(fermented foods)、烘培产品(bakery products)、健康食品(health foods)以及膳食补充品(dietary supplements)。
实施例1.波罗蜜白芯的萃取物的制备
首先,准备波罗蜜的白芯并将它清洗干净供后续萃取之用。接着,取洗净后的波罗蜜白芯并将之均质,然后将均质物以水(2~10∶1~5的液固比)于50~100℃进行萃取0.5~3小时。之后,冷却至室温并经由400目(mesh)的滤网过滤,然后于45~70℃对过滤产物进行减压浓缩,而得到波罗蜜白芯的萃取物。
实施例2.波罗蜜白芯的萃取物在改善骨质疏松症上的效用评估
2.1抑制蚀骨细胞分化试验
首先,由健康捐赠者提供***血液(peripheral blood,PB),然后以15mL的PBS对20mL的***血液进行稀释。接着,添加15mL的聚蔗糖(Ficoll)(GE Healthcare)于具有50mL体积的离心管中,然后添加35mL的经稀释的***血液于聚蔗糖层上,并以400g进行离心40分钟。之后,取出***血液单核细胞(peripheral blood mononuclear cells,PBMCs)并以PBS清洗两次。
接着,将***血液单核细胞分成2组,其中包括1个对照组及1个实验组。以添加有10%胎牛血清(FBS)(Gibco)、1X青霉素(penicillin)/链霉素(streptomycin)(Gibco)、40ng/mL人类核因子κ-B配体受体致活剂(receptor activator of nuclear factorkappa-B ligand,RANKL)(Peprotech,310-01)及25ng/mL人类单核球群落刺激因子(monocyte colony stimulating factor,M-CSF)(Peprotech,300-25)的α-MEM(Gibco,12000-022)作为蚀骨细胞分化培养基来对各组细胞进行培养14天,其中实验组的细胞每3天更换一次添加有0.25mg/mL波罗蜜白芯的萃取物的蚀骨细胞分化培养基,而对照组的细胞每3天更换一次蚀骨细胞分化培养基。
在14天培养之后,产生大量多核蚀骨细胞,接着以1滴Actin GreenTM 488试剂(Thermo,R37110)及赫斯特33342(Hoechst 33342)(Thermo,62249)(1∶20,000倍稀释)进行细胞染色15分钟,然后以荧光显微镜观察蚀骨细胞。本实验的结果显示于图1。
图1是本发明波罗蜜白芯的萃取物在抑制蚀骨细胞分化上的功效的细胞染色图。由图1可见,与对照组相较之下,实验组的细胞多为未分化的单核细胞,而对照组的细胞具有蚀骨细胞特征的多细胞核构造。这个实验结果显示,本发明波罗蜜白芯的萃取物可藉由抑制蚀骨细胞分化来达到改善骨质疏松症的功效。
2.2与蚀骨细胞分化与骨吸收相关的基因表现分析
首先,将依据实施例2.1中的***血液单核细胞分成3组,其中包括2个对照组(亦即对照组1与对照组2)及1个实验组。对照组1以添加有10%胎牛血清(FBS)(Gibco)、1X青霉素(penicillin)/链霉素(streptomycin)(Gibco)的α-MEM(Gibco,12000-022)进行培养,对照组2与实验组以添加有10%胎牛血清(FBS)(Gibco)、1X青霉素(penicillin)/链霉素(streptomycin)(Gibco)、40ng/mL人类核因子κ-B配体受体致活剂(receptor activatorof nuclear factor kappa-B ligand,RANKL)(Peprotech,310-01)及25ng/mL人类单核球群落刺激因子(monocyte colony stimulating factor,M-CSF)(Peprotech,300-25)的α-MEM(Gibco,12000-022)进行培养。3组细胞皆培养于6孔盘上,浓度为106细胞/孔,其中实验组的细胞需额外添加有0.25mg/mL波罗蜜白芯的萃取物,培养24小时后,收取各组细胞培养物并拿来进行基因表现分析。
在本实施例中,用来分析与蚀骨细胞分化相关的基因包括CFOS基因、群落刺激因子1受体(colony stimulating factor 1 receptor,CSF1R)基因,及RANK基因;与蚀骨细胞骨吸收相关的基因包括脾脏酪胺酸激酶(spleen tyrosinase kinase,SYK)基因及抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)基因。
以RNA萃取套组(RNA extraction kit)(Geneaid)对上面所得到的各组细胞培养物进行RNA的萃取。对由此所得到的各组RNA取2,000ng并以III反转录酶(III Reverse Transcriptase)(Invitrogen)将萃取出的RNA反转录为cDNA。接着,以cDNA作为模版(template),并且使用用来扩增目标基因的引物对[包括CFOS、CSF1R、RANK、SYK、TRAP,及RPLP0(作为内部对照组),它们的核苷酸序列显示于下面表1],在StepOne Plus实时PCR***(StepOne Plus Real Time PCR system)(ABI)中利用KAPACYBR FAST qPCR套组(2x)(KAPA Biosystems)来进行定量实时聚合酶链反应(quantitative real-time polymerase chain reaction,以下简称定量实时PCR),俾以对目标基因进行扩增及定量。PCR产物的熔化曲线(melting curve)是在定量实时PCR反应期间进行确认。
表1
目标基因的相对表现量是推导自方程式2-ΔΔCt,并利用RPLP0基因(作为内部对照组)及基准基因的循环阈值及藉由标准偏差来计算相对倍数变化,其中ΔCt=Ct目标基因/基准基因-CtRPLP0,ΔΔCt=ΔCt目标基因-ΔCt基准基因,倍数变化=2-ΔΔCt 平均值。以对照组1的目标基因表现量作为1的比较基准。各组之间的统计学显著差异是藉由单尾史徒登氏t-检定(single tailed Student’s t-test)来决定。本实施例的结果显示于图2及图3。
图2是本发明波罗蜜白芯的萃取物在调控与蚀骨细胞分化相关的基因(包括CFOS基因、CSF1R基因及RANK基因)表现上的功效的数据图;图3是本发明波罗蜜白芯的萃取物在调控与蚀骨细胞骨吸收相关的基因(包括SYK基因及TRAP基因)表现上的功效的数据图。由图2可见,无论是CFOS基因、CSF1R基因,或是RANK基因,与对照组1相较之下,对照组2测得与蚀骨细胞分化相关的基因相对表现量有提升的现象,这表示蚀骨细胞可进行分化,而与对照组2相较之下,实验组测得与蚀骨细胞分化相关的基因相对表现量有降低的现象,这表示波罗蜜白芯的萃取物确实可抑制蚀骨细胞分化。由图3可见,就TRAP基因而言,与对照组1相较之下,对照组2测得与蚀骨细胞骨吸收相关的基因相对表现量有提升的现象,这表示蚀骨细胞可进行骨吸收,而无论是SYK基因或是TRAP基因,与对照组2相较之下,实验组测得与蚀骨细胞骨吸收相关的基因相对表现量有显著的降低,这表示波罗蜜白芯的萃取物确实可抑制蚀骨细胞进行骨吸收。这个实验结果显示,本发明波罗蜜白芯的萃取物可藉由负向调控与蚀骨细胞分化与骨吸收相关的基因表现来达到改善骨质疏松症的功效。
实施例3.波罗蜜白芯的萃取物在心血管保健上的效用评估
在本实施例中,申请人探讨波罗蜜白芯的萃取物可否藉由调控与HDL生成及胆固醇代谢相关的基因表现来达到心血管保健的功效。
首先,以添加有FBS(Gibco,10437-028)、青霉素/链霉素(Gibco,15140122)的DMEM培养基(Gibco,12100-038)培养HepG2细胞于6-孔培养盘,细胞浓度为1×105细胞/孔,接而于37℃进行培养24小时,并移除培养基。
之后,将经培养的细胞分成2组,其中包括1个对照组及1个实验组。将实施例1得到的波罗蜜白芯的萃取物以培养基稀释为具有0.25mg/ml浓度的稀释液,继而添加至实验组的细胞中,至于对照组的细胞则添加培养基。接着,收取各组细胞培养物并拿来进行基因表现分析。
在本实施例中,用来分析与HDL生成及胆固醇代谢相关的基因包括脂蛋白元A-1(apolipoprotein A-1,APA1)基因及ATP结合匣子族A成员1(ATP binding cassettesubfamily A member 1,ABCA1)基因。
以RNA萃取套组对上面所得到的各组细胞培养物进行RNA的萃取。对由此所得到的各组RNA取2,000ng并以III反转录酶将萃取出的RNA反转录为cDNA。接着,以cDNA作为模版,并且使用用来扩增目标基因的引物对[包括APA1、ABCA1,及β-肌动蛋白(β-actin)(作为内部对照组),它们的核苷酸序列显示于下面表2],在StepOne Plus实时PCR***中利用KAPA CYBR FAST qPCR套组(2x)来进行定量实时PCR,俾以对目标基因进行扩增及定量。PCR产物的熔化曲线是在定量实时PCR反应期间进行确认。
表2
目标基因的相对表现量是推导自方程式2-ΔΔCt,并利用β-肌动蛋白基因(作为内部对照组)及基准基因的循环阈值及藉由标准偏差来计算相对倍数变化,其中ΔCt=Ct目标基因/基准基因-Ctβ-肌动蛋白,ΔΔCt=ΔCt目标基因-ΔCt基准基因,倍数变化=2-ΔΔCt 平均值。以对照组的目标基因表现量作为1的比较基准。各组之间的统计学显著差异是藉由单尾史徒登氏t-检定来决定。本实施例的结果显示于图4。
图4是本发明波罗蜜白芯的萃取物在调控与HDL生成及胆固醇代谢相关的基因(包括APA1基因及ABCA1基因)表现上的功效的数据图,由图4可见,无论是APA1基因或是ABCA1基因,与对照组相较之下,实验组测得与HDL生成及胆固醇代谢相关的基因相对表现量有显著的提升。本实施例的结果显示,本发明波罗蜜白芯的萃取物可藉由正向调控与HDL生成及胆固醇代谢相关的基因表现来达到心血管保健的功效。
实施例4.波罗蜜白芯的萃取物在改善热潮红上的效用评估
在本实施例中,申请人探讨波罗蜜白芯的萃取物可否藉由调控与内皮细胞发炎反应与细胞凋亡相关的基因表现来达到改善热潮红的功效。
首先,以添加有LSGS(Gibco,S-003-10)和青霉素/链霉素(Gibco,15140122)的Medium 200培养基(Gibco,M-200-500)培养HUVEC细胞于6-孔培养盘,细胞浓度为1×105细胞/孔,接而于37℃进行培养24小时,并移除培养基。
之后,将经培养的细胞分成2组,其中包括1个对照组及1个实验组。将实施例1得到的波罗蜜白芯的萃取物以培养基稀释为具有0.25mg/mL浓度的稀释液,继而添加至实验组的细胞中,至于对照组的细胞则添加培养基。在历时48小时的作用之后,收取各组细胞培养物并拿来进行基因表现分析。
在本实施例中,用来分析与内皮细胞发炎反应相关的基因包括介白素6(interleukin-6,IL-6)基因及血管细胞黏附分子1(vascular cell adhesion molecule1,VCAM1)基因;与内皮细胞细胞凋亡相关的基因为凋亡蛋白酶8(caspase 8,CASP8)基因。
以RNA萃取套组对上面所得到的各组细胞培养物进行RNA的萃取。对由此所得到的各组RNA取2,000ng并以III反转录酶将萃取出的RNA反转录为cDNA。接着,以cDNA作为模版,并且使用用来扩增目标基因的引物对[包括IL-6、VCAM1、CASP8,及β-肌动蛋白(作为内部对照组),它们的核苷酸序列显示于下面表3],在StepOne Plus实时PCR***中利用KAPA CYBR FAST qPCR套组(2x)来进行定量实时PCR,俾以对目标基因进行扩增及定量。PCR产物的熔化曲线是在定量实时PCR反应期间进行确认。
表3
目标基因的相对表现量是推导自方程式2-ΔΔCt,并利用β-肌动蛋白基因(作为内部对照组)及基准基因的循环阈值及藉由标准偏差来计算相对倍数变化,其中ΔCt=Ct目标基因/基准基因-Ctβ-肌动蛋白,ΔΔCt=ΔCt目标基因-ΔCt基准基因,倍数变化=2-ΔΔCt 平均值。以对照组的目标基因表现量作为1的比较基准。各组之间的统计学显著差异是藉由单尾史徒登氏t-检定来决定。本实施例的结果显示于图5。
图5是本发明波罗蜜白芯的萃取物在调控与内皮细胞发炎反应与细胞凋亡相关的基因(包括IL-6基因、VCAM1基因及CASP8基因)表现上的功效的数据图,由图5可见,无论是IL-6基因、VCAM1基因或是CASP8基因,与对照组相较之下,实验组测得与内皮细胞发炎反应与细胞凋亡相关的基因相对表现量有显著的降低。本实施例的结果显示,本发明波罗蜜白芯的萃取物可藉由负向调控与内皮细胞发炎反应与细胞凋亡相关的基因表现来达到改善热潮红的功效。
实施例5.波罗蜜白芯的萃取物在改善失眠上的效用评估
在本实施例中,申请人探讨波罗蜜白芯的萃取物可否藉由调控与色胺酸代谢相关的基因表现来达到改善失眠的功效。
首先,以添加有FBS(Gibco,10437-028)及青霉素/链霉素(Gibco,15140122)的DMEM培养基(Gibco,12100-038)培养SH-SY5Y细胞于6-孔培养盘,细胞浓度为1×105细胞/孔,接而于37℃进行培养24小时,并移除培养基。
之后,将经培养的细胞分成2组,其中包括1个对照组及1个实验组。将实施例1得到的波罗蜜白芯的萃取物以培养基稀释为具有0.25mg/mL浓度的稀释液,继而添加至实验组的细胞中,至于对照组的细胞则添加培养基。在历时48小时的作用之后,收取各组细胞培养物并拿来进行基因表现分析。
在本实施例中,用来分析与色胺酸代谢相关的基因包括DOPA脱羧酶(DOPAdecarboxylase,DDC)基因及乙酰复合胺O-甲基转移酶类蛋白(Acetylserotonin O-methyltransferase-like protein,ASMTL protein)基因。
以RNA萃取套组对上面所得到的各组细胞培养物进行RNA的萃取。对由此所得到的各组RNA取2,000ng并以III反转录酶将萃取出的RNA反转录为cDNA。接着,以cDNA作为模版,并且使用用来扩增目标基因的引物对[包括DDC、ASMTL,及GAPDH(作为内部对照组),它们的核苷酸序列显示于下面表4],在StepOne Plus实时PCR***中利用KAPACYBR FAST qPCR套组(2x)来进行定量实时PCR,俾以对目标基因进行扩增及定量。PCR产物的熔化曲线是在定量实时PCR反应期间进行确认。
表4
目标基因的相对表现量是推导自方程式2-ΔΔCt,并利用β-肌动蛋白基因(作为内部对照组)及基准基因的循环阈值及藉由标准偏差来计算相对倍数变化,其中ΔCt=Ct目标基因/基准基因-CtGAPDH,ΔΔCt=ΔCt目标基因-ΔCt基准基因,倍数变化=2-ΔΔCt 平均值。以对照组的目标基因表现量作为1的比较基准。各组之间的统计学显著差异是藉由单尾史徒登氏t-检定来决定。本实施例的结果显示于图6。
图6是本发明波罗蜜白芯的萃取物在调控与色胺酸代谢相关的基因(包括DDC基因及ASMTL基因)表现上的功效的数据图,由图6可见,无论是DDC基因或是ASMTL基因,与对照组相较之下,实验组测得与色胺酸代谢相关的基因相对表现量有显著的提升。本实施例的结果显示,本发明波罗蜜白芯的萃取物可藉由正向调控与色胺酸代谢相关的基因表现来提升血清素与褪黑激素的生成,进而达到改善失眠的功效。
综上所述,本发明波罗蜜白芯的萃取物确实可调控CFOS基因、CSF1R基因、RANK基因、SYK基因、TRAP基因、APA1基因、ABCA1基因、IL-6基因、VCAM1基因、CASP8基因、DDC基因、及ASMTL蛋白基因的表现量,并藉由抑制蚀骨细胞分化与骨吸收、促进HDL生成与胆固醇代谢、减少内皮细胞发炎反应与细胞凋亡,及提升血清素与褪黑激素生成,达到改善停经症候群(包括骨质疏松症、心血管疾病、热潮红,及失眠)的功效。
以上所述仅为举例性,而非为限制性者。任何未脱离本发明的精神与范畴,而对其进行的等效修改或变更,均应包含于后附的申请专利范围中。
Claims (13)
1.一种波罗蜜(Artocarpus heterophyllus)白芯的萃取物用于制备一调控CFOS基因、群落刺激因子1受体(colony stimulating factor 1 receptor,CSF1R)基因、RANK基因、脾脏酪胺酸激酶(spleen tyrosinase kinase,SYK)基因、抗酒石酸酸性磷酸酶(tartrateresistant acid phosphatase,TRAP)基因、脂蛋白元A-1(apolipoprotein A-1,APA1)基因、ATP结合匣子族A成员1(ATP binding cassette subfamily A member 1,ABCA1)基因、介白素6(interleukin-6,IL-6)基因、血管细胞黏附分子1(vascular cell adhesionmolecule 1,VCAM1)基因、凋亡蛋白酶8(caspase 8,CASP8)基因、DOPA脱羧酶(DOPAdecarboxylase,DDC)基因、及乙酰复合胺O-甲基转移酶类蛋白(Acetylserotonin O-methyltransferase-like protein,ASMTL protein)基因的表现量的医药品或食品产品的用途。
2.根据权利要求1所述的用途,其特征在于,所述波罗蜜白芯的萃取物是以水或醇类作为萃取溶剂对该波罗蜜白芯进行萃取而制得。
3.根据权利要求2所述的用途,其特征在于,所述萃取溶剂与该波罗蜜白芯的液固比介于2~10∶1~5。
4.根据权利要求2所述的用途,其特征在于,所述萃取的温度介于50℃至100℃。
5.根据权利要求2所述的用途,其特征在于,所述萃取的时间介于0.5至3小时。
6.根据权利要求1所述的用途,其特征在于,所述医药品包含一医药上可接受的载剂。
7.一种波罗蜜(Artocarpus heterophyllus)白芯的萃取物用于制备一改善停经症候群的医药品或食品产品的用途。
8.根据权利要求7所述的用途,其特征在于,所述波罗蜜白芯的萃取物是以水或醇类作为萃取溶剂对该波罗蜜白芯进行萃取而制得。
9.根据权利要求8所述的用途,其特征在于,所述萃取溶剂与该波罗蜜白芯的液固比介于2~10∶1~5。
10.根据权利要求8所述的用途,其特征在于,所述萃取的温度介于50℃至100℃。
11.根据权利要求8所述的用途,其特征在于,所述萃取的时间介于0.5至3小时。
12.根据权利要求7所述的用途,其特征在于,所述停经症候群包括骨质疏松症、心血管疾病、热潮红,及失眠。
13.根据权利要求7所述的用途,其特征在于,所述医药品包含一医药上可接受的载剂。
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