CN115039878A - 一种调节脂肪代谢功能的组合物 - Google Patents
一种调节脂肪代谢功能的组合物 Download PDFInfo
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- CN115039878A CN115039878A CN202110255328.8A CN202110255328A CN115039878A CN 115039878 A CN115039878 A CN 115039878A CN 202110255328 A CN202110255328 A CN 202110255328A CN 115039878 A CN115039878 A CN 115039878A
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- fucoxanthin
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- green tea
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Abstract
本发明公开了一种调节脂肪代谢功能的组合物,其特征在于,所述组合物包含岩藻黄素的提取物和含茶多酚的绿茶提取物,且含岩藻黄素的提取物和含茶多酚的绿茶提取物的重量比为1:6~1:12。本发明的组合物相互作用,可以抑制从食物中吸收脂肪,并抑制体内脂肪分化及合成,从而调节机体的脂质代谢。
Description
技术领域
本发明涉及食品、保健食品、药品技术领域,尤其涉及一种调节脂肪 代谢功能的组合物。
背景技术
肥胖主要是由于能量摄入过剩,促进体内脂肪合成,从而导致体内脂 肪过度堆积。该报告还表明少体力活动和不健康的生活习惯也与超重、肥 胖密切相关。除此之外,肥胖还易提高糖尿病、高血压及血脂异常等患病 风险。因此科学合理的营养治疗联合运动仍是目前最有效、最安全的降低 肥胖率的干预手段。
从细胞生物学的角度来说,多功能干细胞分化成可定向分化的脂肪前 体细胞,再由脂肪前体细胞分化为成熟的脂肪细胞。脂肪在体内形成后, 将会在成熟的脂肪细胞间质中囤积,脂肪囤积的越多,造成的脂肪细胞体 积越大。
除了通过减少能量摄入或者增加能量消耗外,抑制前脂肪细胞分化增 殖、脂肪生成以及促进脂解作用以及脂肪氧化也都为改善肥胖的可能机理。
岩藻黄素分子式为C42H58O6,主要由褐藻、硅藻、定鞭藻、金藻、针胞 藻等海洋藻类经提取得来。岩藻黄素有很长的共扼双键,并且其分子结构 中具有特殊的丙二烯和5,6-环氧烷,这种特殊结构决定了岩藻黄素具有许 多重要的生物学功能。现有研究表明岩藻黄素具有燃烧脂肪,增加能量消 耗的作用。岩藻黄素是通过提高解偶联蛋白UCP1(UncouplingProtein 1) 的表达来实现这一作用。UCP1是一类位于线粒体内膜上的载体,可以将H+从线粒体内膜渗漏到线粒体基质中,减少ATP的合成并产生热能。UCP1是 控制能量稳态和体重的重要指标之一,通过上调UCP1的表达将会成为缓解 肥胖一种有效的干预手段。通过实验发现岩藻黄素提高了高脂饮食诱导的 肥胖小鼠体内脂肪组织中的UCP1的表达,增强了脂肪的产热功能,进而有 效的起到了减脂的作用。
也有研究表明,岩藻黄素在3T3-L1细胞(小鼠胚胎成纤维细胞)的脂 肪组织分化过程中,可以抑制细胞内的脂质聚积和甘油-3-磷酸脱氢酶的活 性。岩藻黄素及其代谢产物岩藻黄醇在前体脂肪细胞3T3-L1中可降低过氧 化物酶增殖体激活受体((PPARy)的表达,从而抑制其向成熟脂肪细胞分化。 这说明岩藻黄素除了可以上调脂肪组织中UCP1的表达之外还可抑制脂肪前 体细胞定向分化。
但研究表明岩藻黄素需要连续服用8周起才开始显示出减脂作用,起 效时间较长,短期内无法见效,容易让使用者难以坚持,提前放弃服用。 而岩藻黄素抑制脂肪从肠道吸收、抑制成熟脂肪细胞对葡萄糖的吸收和利 用作用虽然较强,但其促进脂肪细胞的凋亡作用却较弱。
发明内容
本发明的目的在于克服现有技术的缺陷,提供一种既能快速改善脂质代谢, 抑制脂肪细胞分化,抑制脂肪细胞对葡萄糖的利用,又能促进成熟脂肪细 胞凋亡和抑制脂肪吸收作用的组合物。
本发明的技术方案如下:
一种调节脂肪代谢功能的组合物,其特征在于,所述组合物包含含岩 藻黄素的提取物和含茶多酚的绿茶提取物,且两者的重量比为1:6–1:12。
岩藻黄素的主要生理学功能为:激活线粒体内膜上的UCP-1(解偶联蛋 白-1)开启氢离子通道,在不产生ATP的前提下使脂肪细胞持续产热,以 达到消耗脂肪的目的;茶多酚的主要生理学功能为:通过上调组织中CPT1a 和ACOX1促进脂肪酸的β氧化,并通过激活PGC1α信号通路增强线粒体功 能,以此提高脂肪供能比率。因此本专利将首次以含有茶多酚的绿茶提取 物弥补岩藻黄素消耗脂肪却无法提供ATP的缺点,同时岩藻黄素通过调节脂肪产热,防止ATP过度生成造成的自由基积累。
岩藻黄素有着加速脂肪燃烧分解的作用。含有茶多酚的绿茶提取物可 以通过激活AMPK信号通路抑制脂肪及胆固醇的生成。此外含有茶多酚的绿 茶提取物还可以抑制脂肪细胞的增殖和分化,加速脂肪细胞凋亡,并抑制 由肥胖造成的炎症因子上调。因此本专利首次将岩藻黄素结合绿茶提取物 可以综合从抑制脂肪吸收、抑制脂肪生成,加速脂肪分解等途径实现减脂 的目的。
作为对上述技术方案的进一步改进,含岩藻黄素的提取物和含茶多酚 的绿茶提取物的重量比为1:7~1:10。
作为对上述技术方案的进一步改进,所述含岩藻黄素的提取物来源包 括植物来源、微生物来源。
作为对上述技术方案的进一步改进,含茶多酚的绿茶提取物的茶多酚 含量为20%-80%,含岩藻黄素的提取物的岩藻黄素含量为0.5%-10%。
作为对上述技术方案的进一步改进,所述含茶多酚的绿茶提取物经过 粉碎、水或乙醇提取、浓缩、干燥工艺制备而成,所述含岩藻黄素的提取 物经过粉碎、水或乙醇提取、浓缩、干燥工艺制备而成。
作为对上述技术方案的进一步改进,所述组合物为口服制剂。
作为对上述技术方案的进一步改进,所述口服制剂为粉剂、颗粒剂、 硬胶囊、软胶囊、片剂、软糖、口服溶液或乳化剂。
作为对上述技术方案的进一步改进,所述组合物还包括可用于药品、 食品中的辅料或添加剂,所述辅料或添加剂为甜味剂、酸调节剂、填充剂、 矫味剂、着色剂、抗氧化剂、增稠剂、稳定剂、乳化剂、抗结剂、助流剂、 润滑剂中的一种或多种。
本发明具有以下技术效果:
1.本发明的组合物既能改善脂质代谢,抑制脂肪分化,增强抑制脂肪 吸收作用,又能抑制脂肪生成和加速脂肪分解。
2.本发明组合物成分和功效明确,适用范围广,无毒副作用。
3.本发明组合物制备操作简单,成本低。
4.本发明适用剂型广,便于应用。
附图说明
图1显示不同组别对脂肪细胞分化的影响,其中A为空白组;B为阳性参照 组;C为含岩藻黄素提取物组(50μg/mL);D为含茶多酚的绿茶提取物组 (50μg/mL);E为组合配方组(50μg/mL)。
图2显示不同组别的油红O染色吸光度值。
图3显示不同组别对脂肪细胞葡萄糖利用度的影响。
具体实施方式
下面结合具体实施例对本发明作进一步说明,但本发明并不限于以下 实施例。
实施例1:组合物配置
将含岩藻黄素的提取物和含茶多酚的绿茶提取物按照下表1所示重量 比例进行混合均匀。
表1:组合物配置
实施例2:脂肪酶活性抑制试验
脂肪的吸收利用需要在脂肪酶的作用下分解为甘油和脂肪酸,再经由 小肠吸收进入血液。因此抑制脂肪酶的活性在一定程度上可以起到抑制脂 肪吸收的作用。通过测定体外脂肪酶活性抑制程度来评价抑制脂肪吸收的 能力。
分别测定了含岩藻黄素的提取物、含茶多酚的绿茶提取物体外脂肪酶 抑制活性,以半抑制浓度(IC50)表示。再将两物质以不同比例复配测定其 IC50,以相互作用指数γ评价协同作用的程度。
试剂制备
1.pNP laurate溶液配置
a.称取41mg乙酸钠,溶于100ml蒸馏水,加1mL Triton X-100,得 5mM乙酸钠溶液(含1%Triton X-100)。
b.称取80mg 4-硝基苯基十二酸酯溶于100ml 5mM乙酸钠溶液(含1% Triton X-100),于65℃下搅拌15min使其混合均匀,获得pNP laurate (十二酸酯)溶液(需要避光保存)。
2.0.1M Tris-HCl(pH 8.2)缓冲液制备
3.脂肪酶溶液制备
转移50μL脂肪酶至20mL容量瓶,加入0.1M Tris-HCl(pH8.2)缓冲 液,混合均匀,获得脂肪酶溶液。
4.样品溶液制备
将样品用0.1M Tris-HCl(pH 8.2)缓冲液稀释到相应的浓度。
实验步骤
1.样品脂肪酶抑制活性测定
所有组先加入200μL Tris-HCl缓冲液。
样品组:100μL不同浓度的样品溶液,100μL脂肪酶溶液加至EP管。
样品空白组:100μL不同浓度的样品溶液,100μL Tris-HCl缓冲液 加至EP管。
阴性对照组:100μL Tris-HCl缓冲液,100μL脂肪酶溶液加至EP管。
空白对照组:200μL Tris-HCl缓冲液加至EP管。
所有EP管置于37℃水浴中反应15min。
所有EP管加入200μL pNP laurate溶液于37℃下反应30min。
反应之后,将所有管置于95℃水浴中5min终止反应。降至室温后, 所有管于6000rpm下离心3min。转移200μL上清液至96孔板中,于405nm 波长下测定吸光度值。
数据分析
胰脂肪酶抑制率计算公式如下:
胰脂肪酶抑制率(%)=[(A阴性-A空白)-(A样品-A样空)/(A阴 性-A空白)]×100%
式中:A阴性为阴性对照组测得吸光度,A空白为空白对照组测得吸光 度,A样品为样品组测得吸光度,A样空为样品空白组测得吸光度。
再由不同浓度的样品的抑制率求出IC50值。
结果显示:
含岩藻黄素的提取物(岩藻黄素1%)IC50:5.9mg/mL
含茶多酚的绿茶提取物(茶多酚60%,以儿茶素类总量计)IC50: 0.75mg/mL
含茶多酚的绿茶提取物(茶多酚80%,以儿茶素类总量计)IC50:0.90mg/mL
含茶多酚的绿茶提取物(茶多酚98%,以儿茶素类总量计) IC50:1.15mg/mL
含茶多酚的绿茶提取物(茶多酚20%,以儿茶素类总量计)IC50: 0.89mg/mL
EGCG(98%,表示没食子儿茶素没食子酸酯)IC50:15.89mg/mL
表2:脂肪酶活性抑制试验结果(含岩藻黄素的提取物中岩藻黄素1%, 含茶多酚的绿茶提取物中茶多酚60%(以儿茶素类总量计))
表3:脂肪酶活性抑制试验结果
含茶多酚的绿茶提取物(茶多酚60%,以儿茶素类总量计)IC50: 0.75mg/mL。
含岩藻黄素的提取物(岩藻黄素0.2%)IC50:10.32mg/mL。
实施例3:对脂肪细胞分化的影响
3.1 3T3-L1细胞诱导分化
将3T3-L1前脂肪细胞接种于培养板,用含10%胎牛血清的高糖DMEM培 养液在37℃、5%CO2条件下培养,待细胞长满至80%~90%、用胰酶消化 下细胞,制成细胞悬液(第0天)。
将细胞分为空白组(加入诱导剂)、含岩藻黄素提取物组(加入诱导剂和 含岩藻黄素提取物(50μg/mL))、含茶多酚的绿茶提取物组(加入诱导 剂和含茶多酚的绿茶提取物(50μg/mL))、组合配方组(加入诱导剂和含茶 多酚的绿茶提取物和含岩藻黄素提取物(重量比10:1,50μg/mL))、阳 性参照组(加入诱导剂和氯化锂)。
同时,高糖DMEM培养48h,然后换含终质量浓度10μg/mL胰岛素的 10%胎牛血清高糖DMEM培养液继续培养48h,每2d换液1次,诱导分化第 9天后结束实验。
诱导剂配方:将3-异丁基-1-甲基黄嘌呤用乙醇配制成111.1mg/mL (0.5mol/L),将地塞米松用乙醇配制成1mg/mL(2.5mmol/L),将10mg胰岛 素溶于1mL 0.01mol/L HCl中。诱导分化剂MDI的配方为10%胎牛血清, 0.5mmol/L 3-异丁基-1-甲基黄嘌呤,1μmol/L地塞米松,5μg/mL胰岛 素。
3.2油红O染色
处理后的细胞,弃去培养基并用冷PBS清洗细胞两遍;用4%多聚甲醛 溶液固定细胞60min;固定好的细胞用油红O染液试剂盒进行染色;用60% 的异丙醇清洗细胞3次除去未着色的油红O染液,显微镜下进行拍照观察; 观察结束后,裂解液(含4%NP-40)裂解细胞并测定在490nm处OD值。
3.3数据分析
根据测出各组OD值后,比较各样品组与对照组之间有没有显著性差异, 若样品组与诱导分化组有显著性差异,则证明样品对脂肪分化有抑制作用。
实验结果如图1、2所示。油红O染色OD值相比较,组合配方组与空 白组、含岩藻黄素的提取物组、含茶多酚的绿茶提取物组相比明显减小, 存在显著性差异(P<0.01)。说明含岩藻黄素的提取物与含茶多酚的绿茶 提取物以特定比例复配后可增强抑制脂肪分化的效果。
实施例4:对脂肪细胞葡萄糖利用度的影响
4.1 3T3-L1细胞诱导分化
将3T3-L1前脂肪细胞接种于培养板,用含10%胎牛血清的高糖DMEM培 养液在37℃、5%CO2条件下培养,待细胞长满至80%~90%、用胰酶消化 下细胞,制成细胞悬液(第0天)。将细胞以105每孔的密度接种在96孔板 中,待细胞长至融合加入分化诱导剂诱导分化,2d后换液加入10μg/mL胰 岛素的10%胎牛血清高糖DMEM培养液继续培养48h,每2d换液1次,诱导 分化第9天后,将细胞分为空白组、含岩藻黄素提取物组(加入诱导剂和含 岩藻黄素提取物)、含茶多酚的绿茶提取物组(加入诱导剂和含茶多酚的 绿茶提取物)、组合配方组(加入诱导剂和含茶多酚的绿茶提取物:含岩藻 黄素的提取物=10:1)、阳性参照组(加入诱导剂和氯化锂),再用高糖DMEM 培养24h,然后用葡萄糖氧化酶法测量葡萄糖浓度。
诱导剂配方:将3-异丁基-1-甲基黄嘌呤用乙醇配制成111.1mg/mL (0.5mol/L),将地塞米松用乙醇配制成1mg/mL(2.5mmol/L),将10mg胰岛 素溶于1mL 0.01mol/L HCl中。诱导分化剂MDI的配方为10%胎牛血清, 0.5mmol/L 3-异丁基-1-甲基黄嘌呤,1μmol/L地塞米松,5μg/mL胰岛 素。
4.2葡萄糖利用度检测
1.吸取培养基,1000rpm离心5min,取上清液;
2.采用GOP-POD法试剂盒检测培养基上清中葡萄糖含量;
3.诱导分化过程中细胞的葡萄糖摄取量=空白孔葡萄糖含量均值-细胞 接种孔葡萄糖含量均值;
4.3检测结果
检测结果如图3所示,组合配方组与空白组、含岩藻黄素的提取物组相比 均明显减少葡萄糖的利用,存在显著性差异(P<0.01)。说明含岩藻黄素 的提取物与含茶多酚的绿茶提取物以特定比例复配后可抑制脂肪细胞对葡 萄糖的利用度而避免脂肪堆积。
显然,本发明的上述实施例仅仅是为清楚地说明本发明所作的举例, 并非是对本发明实施方式的限定。对于所属领域的普通技术人员来说,在 上述说明的基础上还可以作出其他不同形式的变化或变动。这里无法对所 有的实施方式予以穷举。凡是属于本发明的技术方案所引申出的显而易见 的变化或变动仍处于本发明保护范围之列。
Claims (8)
1.一种调节脂肪代谢功能的组合物,其特征在于,所述组合物包含含岩藻黄素的提取物和含茶多酚的绿茶提取物,含岩藻黄素的提取物和含茶多酚的绿茶提取物的重量比为1:6~1:12。
2.根据权利要求1所述的组合物,其特征在于,含岩藻黄素的提取物和含茶多酚的绿茶提取物的重量比为1:7~1:10。
3.根据权利要求1所述的组合物,其特征在于,所述含岩藻黄素的提取物来源包括植物来源、微生物来源。
4.根据权利要求1所述的组合物,其特征在于,含茶多酚的绿茶提取物的茶多酚含量为20%-80%,含岩藻黄素的提取物的岩藻黄素含量为0.5%-10%。
5.根据权利要求4所述的组合物,其特征在于,所述含茶多酚的绿茶提取物经过粉碎、水或乙醇提取、浓缩、干燥工艺制备而成;所述含岩藻黄素的提取物经过粉碎、水或乙醇提取、浓缩、干燥工艺制备而成。
6.根据权利要求1所述的组合物,其特征在于,所述组合物为口服制剂。
7.根据权利要求6所述的组合物,其特征在于,所述口服制剂为粉剂、颗粒剂、硬胶囊、软胶囊、片剂、软糖、口服溶液或乳化剂。
8.如权利要求1所述的组合物,其特征在于,所述组合物还包括可用于药品、食品中的辅料或添加剂,所述辅料或添加剂为甜味剂、酸调节剂、填充剂、矫味剂、着色剂、抗氧化剂、增稠剂、稳定剂、乳化剂、抗结剂、助流剂、润滑剂中的一种或多种。
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