CN114794435B - 一种高营养密度指数混合坚果组合物及其应用 - Google Patents
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Abstract
本发明涉及一种高营养密度指数混合坚果组合物及其应用,属于食品加工技术领域。针对现有技术中坚果等应用于缓解高胆固醇血症或胰岛素抵抗的食品或保健品时,坚果品种单一,缺乏基于营养学科学搭配混合食物依据,以及参与体内氧化还原反应与能量代谢的微量营养素体系组成不完善,导致的缓解高胆固醇血症或胰岛素抵抗效果不佳的技术问题,本申请提供一种高营养密度指数混合坚果组合物,以高营养密度指数为基础,根据坚果的营养素成分特点优化营养密度计算方法,得出最高营养密度下的混合坚果果干配比,通过高脂肪饮食诱导代谢紊乱动物模型,能够缓解高脂饮食诱导的肥胖和体重的升高、降低血液总胆固醇、改善肝脏脂肪累积、调节糖代谢紊乱和胰岛素抵抗。
Description
技术领域
本发明属于食品加工技术领域,具体地,涉及一种高营养密度指数混合坚果组合物及其应用。
背景技术
随着肥胖症的流行,代谢综合征、非酒精性脂肪肝和2型糖尿病等相关疾病的患病率也在上升。胰岛素抵抗在所有这些疾病的发病机制中起着至关重要的作用,胰岛素抵抗是胰岛素作用的靶器官对胰岛素作用的敏感性下降,即正常剂量的胰岛素产生低于正常生物学效应的一种状态,对胰岛素抵抗的治疗成为防治2型糖尿病及其他代谢紊乱疾病的关键。高胆固醇血症是心血管疾病的最重要的危险因素。积极的诊断和管理以改善胰岛素敏感性并降低过多的有害血脂是必要的,从食物入手研发有效缓解胰岛素抵抗和高胆固醇血症的食物配方对于糖脂代谢紊乱的控制具有重要意义。
大量国外流行病学研究证明,坚果摄入能降低患心脑血管疾病的风险。越来越多的证据也表明食用坚果可以通过降低氧化应激来预防心血管疾病,炎症和改善内皮功能。中国专利申请公布号CN108294231A,发明名称为“适合糖尿病患者食用的营养主食”,公开的该营养主食包括谷物粉、谷物胚芽粉、食源性蛋白粉、坚果类种子粉及葫芦科蔬菜粉。该营养主食将谷物胚芽粉、坚果类种子粉及葫芦科蔬菜粉这三类营养物质协同添加,对于抑制血糖升高有着极佳的表现。特别是将坚果类种子粉和葫芦科蔬菜粉同用,有利于进一步改善人体血糖和胰岛素的平衡,对于II型糖尿病有很好的保健作用。人群对坚果的摄入往往是多样的,但大多数研究都是对单一的坚果品种进行研究,混合坚果缓解胰岛素抵抗作用的研究较少,且缺乏基于营养学科学搭配混合食物依据。
基于宏观和微观营养素以及热量的营养成分分析模型可以为创建更健康的饮食提供科学支持,富含营养素食物指数(NRF)是衡量营养密度的新工具,是饮食建议和营养指导综合系统的组成部分,NRF提供了一个有效的指标来评估混合食物的营养素密度,应用NRF算法得出的高指数比例配方是营养密集型的。目前的变体,即总富含营养素食物指数(NRF9.3),是基于9种鼓励营养素的日值百分比之和减去3种限制营养素的最大推荐值百分比之和,所有日值按每100千卡计算,上限为100%。但是,不同食物的营养素构成不同,只局限于目前的NRF9.3计算营养密度调整混合食物搭配比例是不足的。欧洲专利公开号EP2303252A1,发明名称为“Compounds for the treatment of metabolic syndrome andinsulin resistance”,公开了一种包含鞣花酸部分的化合物,例如鞣花酸本身、鞣花单宁和鞣花酸糖苷,以及包含它们的植物提取物,用于治疗或预防代谢综合征或非糖尿病胰岛素抵抗。是浆果、水果和坚果种类,优选覆盆子、黑莓、云莓、北极荆棘、草莓、石榴、核桃或橡树的提取物。
综上,如何根据坚果和果干的营养素成分特点优化营养密度计算方法,得出最高营养密度下的混合坚果果干配比,并纳入坚果富含的特征营养素,使得混合坚果产品能够缓解高脂饮食诱导的肥胖和体重的升高、降低血液总胆固醇、改善肝脏脂肪累积、调节糖代谢紊乱和胰岛素抵抗是一个亟待解决的问题。
发明内容
1、要解决的问题
针对现有技术中坚果等应用于缓解高胆固醇血症或胰岛素抵抗的食品或保健品时,坚果品种单一,缺乏基于营养学科学搭配混合食物依据,以及参与体内氧化还原反应与能量代谢的微量营养素体系组成不完善,导致的缓解高胆固醇血症或胰岛素抵抗效果不佳的技术问题,本申请提供一种高营养密度指数混合坚果组合物,以NRF指数为基础,根据坚果和果干的营养素成分特点优化营养密度计算方法,得出最高营养密度下的混合坚果果干配比,并通过高脂肪饮食诱导代谢紊乱动物模型,能够缓解高脂饮食诱导的肥胖和体重的升高、降低血液总胆固醇、改善肝脏脂肪累积、调节糖代谢紊乱和胰岛素抵抗。
2、技术方案
为达到上述目的,提供的技术方案为:
本申请的一种高营养密度指数混合坚果组合物,包括混合坚果和混合果干;所述混合坚果和混合果干的比例在线性规划计算中单位热量的总富含营养素食物指数最大。
进一步地,所述总富含营养素指数的计算公式为:
TNRF=NRF10.3×X+NRF8.3×Y,
其中NRF10.3为所述混合坚果富含营养素指数,
NRF8.3为所述混合果干的富含营养素指数,
X和Y分别为所述混合坚果和所述混合果干的质量占比,X+Y=1;
所述富含营养素指数的计算公式为:
NRFn.3=NR-LIM,
其中
nutrienti是第i种鼓励营养素的含量,DVi是该营养素的每日需要量参考值,MRVi是第i种限制营养素的最大推荐值。
进一步地,所述混合坚果和混合果干的富含营养素食物指数的计算均包括9种鼓励营养素和3种限制营养素。
进一步地,所述混合坚果的9种鼓励营养素为:维生素A、维生素B2、维生素E、铁、镁、锌、鞣花酸、亚麻酸、精氨酸和酪氨酸;所述混合坚果的3种限制营养素为:支链氨基酸、蛋氨酸和饱和脂肪酸。
进一步地,所述混合果干的9种鼓励营养素为:维生素A、维生素B2、维生素C、镁、鞣花酸、花青素、精氨酸和酪氨酸;所述混合果干的3种限制营养素为:支链氨基酸、蛋氨酸和果糖。
进一步地,按重量份计算,所述混合坚果为核桃、巴旦木、腰果和榛子;所述混合果干为黑加仑干、蓝莓干和蔓越莓干。
进一步地,所述混合坚果和混合果干的重量比为5~8:2~5。
优选的,所述混合坚果和混合果干的重量比为7:3。
进一步地,按重量份计算,所述混合坚果为核桃30~60%、巴旦木3~12%、腰果5~15%和榛子2~10%;所述混合果干为黑加仑干5~15%、蓝莓干5~10%和蔓越莓干5~15%。
优选的,按重量份计算,所述混合坚果为核桃41.5%、巴旦木9%、腰果11.5%和榛子7%;所述混合果干为黑加仑干11%、蓝莓干9%和蔓越莓干11%。
一种高营养密度指数混合坚果组合物的应用,将所述高营养密度指数混合坚果组合物应用于缓解高胆固醇血症或胰岛素抵抗。
优选的,所述应用包括降低血液总胆固醇、缓解肝脏脂肪累积或改善机体的胰岛素抵抗。
进一步地,将所述高营养密度指数混合坚果组合物应用于食品或保健品中。
优选的,所述食品为麦片、乳制品、豆制品、果蔬制品或饮料。
进一步地,所述食品或保健品中还包含常规辅料。
优选的,所述常规辅料包括但不限于填充剂、食用香精、甜味剂、粘合剂、崩解剂、润滑剂、抗酸剂、以及营养强化剂中的一种或多种。
3、有益效果
采用本发明提供的技术方案,与已有的公知技术相比,具有如下有益效果:
(1)本发明的一种高营养密度指数混合坚果组合物,包括混合坚果和混合果干,混合坚果和混合果干的比例在线性规划计算中单位热量的总富含营养素食物指数最大,在制备预防和/或治疗血脂代谢紊乱和胰岛素抵抗的产品中具有巨大的应用前景。以NRF指数为基础,根据坚果和果干的营养素成分特点优化营养密度计算方法,得出最高营养密度下的混合坚果果干配比,并通过高脂肪饮食诱导代谢紊乱动物模型,能够缓解高脂饮食诱导的肥胖和体重的升高、降低血液总胆固醇、改善肝脏脂肪累积、调节糖代谢紊乱和胰岛素抵抗。
(2)本发明的一种高营养密度指数混合坚果组合物的应用,将高营养密度指数混合坚果组合物应用于食品或保健品中,动物实验表明:本发明的组合物可以缓解高脂肪饮食诱导引起的肥胖;可以降低高脂肪饮食诱导小鼠的血液总胆固醇水平,改善肝脏脂肪累积;可以改善高脂肪饮食诱导小鼠的糖代谢紊乱,缓解胰岛素抵抗。
附图说明
图1为高富含高营养密度指数混合坚果喂养小鼠肝脏和脂肪的病理切片。A.油红O染色的肝脏切片(比例尺:50μm);B.H&E染色的肝脏切片(比例尺:50μm);C.H&E染色的附睾脂肪切片(比例尺:50μm);D.肝组织油红O切片的脂滴率;E.附睾脂肪细胞面积。结果为平均值±标准误,与LFD组相比,*P<0.05,**P<0.01,***P<0.001;与HFD组相比,#P<0.05,##P<0.01,###P<0.001。
图2为高富含高营养密度指数混合坚果对脂代谢的影响。A.血浆总胆固醇;B.总甘油三酯;C.低密度脂蛋白胆固醇;D.高密度脂蛋白胆固醇。结果为平均值±标准误,与LFD组相比,*P<0.05,**P<0.01,***P<0.001;与HFD组相比,#P<0.05。
图3为高富含高营养密度指数混合坚果对糖代谢的影响。A.口服葡萄糖耐量试验后的血糖反应曲线;B.口服葡萄糖耐量试验的血糖曲线下面积;C.胰岛素抵抗指数,胰岛素抵抗指数=空腹血糖水平(mmol/L)×空腹胰岛素水平(mU/L)/22.5。结果为平均值±标准误,与LFD组相比,*P<0.05,**P<0.01,***P<0.001;与HFD组相比,#P<0.05,##P<0.01,###P<0.001。
具体实施方式
为进一步了解本发明的内容,结合实施例对本发明作详细描述。
本混合坚果果干组合物的配方设计中鼓励营养素的选择遵循FDA对健康食品的定义,保留原始NRF中的鼓励营养素:维生素A、C、E、钙和镁,这些营养素不仅是维持机体正常生理功能不可或缺的,而且在参与抗氧化过程发挥重要作用。在此基础上补充纳入维生素B2,锌和酪氨酸,进一步完善参与体内氧化还原反应与能量代谢的微量营养素体系组成。此外分别纳入坚果和果干富含的特征营养素:不饱和脂肪酸和花青素。本组合物配方独特之处在于从坚果混合物发挥预防高胆固醇血症和胰岛素抵抗作用的应用着手,纳入鞣花酸、精氨酸。鞣花酸是在浆果和坚果中发现的天然抗氧化剂多酚,大量研究证实鞣花酸能够降低空腹血糖水平,改善胰岛素抵抗,通过下调烟酰胺二磷酸腺苷氧化酶亚单位(p47-phox)和上调NRF2的表达,增加磷酸化的Akt(Ser 473)水平,激活了肝脏中的胰岛素信号通路。此外,鞣花酸能够减少低氧诱导因子在肝脏表达,从而缓解缺氧和肝脏脂肪变性。精氨酸能够降低高脂肪饮食引起的炎性细胞因子TNF-α水平,这种细胞因子会抑制胰岛素在脂肪组织、肝脏和肌肉中的作用,通过抑制酪氨酸的磷酸化和增加胰岛素受体(IRS-1)的丝氨酸磷酸化来增加胰岛素抵抗,还直接影响葡萄糖的代谢,减少肌肉中GLUT-4的表达,从而导致胰岛素受体活性的降低。
限制营养素的选择首先分别纳入坚果和果干富含的且对营养价值发挥影响较大的饱和脂肪酸和果糖。此外从坚果混合物发挥预防高胆固醇血症和胰岛素抵抗作用的应用着手,纳入支链氨基酸和蛋氨酸。与其他脂质相比,支链氨基酸与胰岛素敏感性及胰岛素抵抗指数的关联性最强。mTOR是氨基酸和胰岛素之间串联的一个核心信号媒介。高浓度的支链氨基酸会持续激活mTORC1,其中亮氨酸直接与SESTRIN 2结合,破坏了与GATOR2的相互作用,并激活mTORC1,通过IRS-1的磷酸化导致胰岛素信号传导受阻,诱导胰岛素抵抗。此外蛋氨酸的限制能够激活棕色脂肪和诱导白色脂肪组织褐变,增加能量消耗,并且激活FGF21-脂联素轴,增加FGF21循环,FGF21能抑制由胰岛素抵抗引起的mTORC1活性的增加,增加肝脏胰岛素敏感性,促进肝脏糖原储存来增加葡萄糖的利用,从而调节全身葡萄糖代谢的动态平衡,进而改善外周胰岛素敏感性。
根据坚果和果干的营养素成分特点,基于以上营养素的鼓励和限制对胰岛素抵抗和高胆固醇血症的影响,优化NR算法,通过线性规划计算使混合坚果和果干的单位热量的TNRE达到最大,得出混合坚果果干的高营养素植物指数配方,其作为缓解高胆固醇血症和胰岛素抵抗的应用价值更高。
实施例1
1.高富含高营养密度混合坚果配方的计算
对混合坚果中4种坚果和3种果干进行营养成分检测,测得的各纳入营养素含量,其中坚果纳入10种鼓励营养素:维生素A、维生素B2、维生素E、铁、镁、锌、鞣花酸、亚麻酸、精氨酸、酪氨酸;3种限制营养素:支链氨基酸、蛋氨酸、饱和脂肪酸,果干纳入8种鼓励营养素:维生素A、维生素B2、维生素C、镁、鞣花酸、花青素、精氨酸、酪氨酸;3种限制营养素:支链氨基酸、蛋氨酸、果糖。
测定混合坚果果干中各营养素的含量,结果如表1-4所示,并计算总富含营养素指数(TNRF),其计算公式为:
TNRF=NRF10.3×X+NRF8.3×Y (1)
NRFn.3=NR-LIM (2)
X+Y=1 (3)
其中NRF10.3为混合坚果富含营养素指数;NRF8.3为混合果干的富含营养素指数;X和Y分别为混合坚果和混合果干的质量占比。NR富含营养素指数的计算公式为,其中nutrienti是第i种鼓励营养素的含量,DVi是该营养素的每日需要量参考值,MRVi是第i种限制营养素的最大推荐值。
根据消费行为调查,当混合坚果和混合果干比例为0.7和0.3时,混合坚果的接受度最高,因此,设定X为0.7,Y为0.3,以指标营养测量数据为基础,采用线性规划的NRF模型,以TNRF指数最高、总能量最低为指标进行配方优化,获得优化的最优混合坚果配方为:核桃41.5%、杏仁9%、腰果11.5%、榛子7%、黑加仑11%、蓝莓9%、蔓越莓11%,优化配方(MN3)的TNRF值为4.15(表5)。
同时根据TNRF公式计算了目前国际上报道较多的无果干混合坚果配方(MN1)的TNRF为3.64。我国市场上常见的一款含果干混合坚果配方(MN2)的TNRF为3.46(表5)。
表1坚果鼓励营养素含量
注:结果为平均值±标准差
表2坚果限制营养素含量
注:结果为平均值±标准差
表3果干鼓励营养素含量
注:结果为平均值±标准差
表4果干限制营养素含量
注:结果为平均值±标准差
表5三种混合坚果配方的总高营养密度指数
MN2和MN3的混合坚果和混合果干的比例为0.7和0.3。
实施例2
1.动物饲养与分组
选取6周龄雄性C57BL/6小鼠(SPF级,购于北京斯贝福实验动物技术有限公司);给料2-3天/次;给水7天/次。预饲养1周后,按体重将小鼠随机分为6组,每组12只,即低脂肪饮食组(10%脂肪,LFD)、高脂肪饮食组(45%脂肪,HFD)、高脂+无果干混合坚果(无果干配方,HFD+MN1)、高脂+低TRNF混合坚果/果干组合物(HFD+MN2)、高脂+高TRNF混合坚果/果干组合物(HFD+MN3)、高脂+单一核桃(HFD+walnut)、高脂+鞣花酸组(HFD+EA),混合坚果添加量根据《中国居民膳食指南2016》推荐每日摄入10g坚果,按成人每日能量需要量(2700kcal)换算为相应的小鼠饲料占比,鞣花酸添加量为300mg/kg饲料。用各组实验饲料饲喂12周,每周测量小鼠体重、摄食量和饮水量。在研究结束前一周进行口服葡萄糖耐量试验。实验结束时,小鼠被禁食6h,处死前测量血糖水平,后采集血液及各组织。
2.口服葡萄糖耐量试验
第11周,每组随机选取6只小鼠,实验前一天对各组小鼠断食不断水,以400mg/mL葡萄糖溶液灌胃,灌胃量为0.1mL/20g,采用尾部取血方法,使用血糖仪分别测定各组小鼠空腹、15分钟、30分钟、60分钟以及90分钟的血糖值,绘制曲线并计算曲线下面积。
3.生化指标测定与分析
在实验结束后,小鼠麻醉处死,收集血样并在1,500×g;4℃下离心10min,参照商业化试剂盒说明书进行总胆固醇(TC)、总甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、胰岛素水平的检测。
用4%多聚甲醛固定新鲜的肝脏和睾丸脂肪组织,并嵌入石蜡中。准备5μm厚的组织切片进行苏木精和伊红(H&E)以及油红O染色。在正置的光学显微镜下进行病理组织学观察。
使用SPSS软件20.0版进行单因素方差分析(ANOVA)和Tukey多重比较检验进行统计学分析,p<0.05表示在统计学意义上具有差异。所有数据以平均值±标准误表示,使用GraphPad Prism 8.0进行数据绘图。
4.结果与分析
4.1高富含高营养密度混合坚果对小鼠体重变化的影响
如表1所示,饲喂低脂肪饲料的小鼠比饲喂高脂肪饲料的小鼠消耗更多的食物(P<0.05)。与LFD组相比,HFD组的小鼠体重明显增加(P<0.05)。与HFD组相比,三个混合坚果组的体重略低,没有统计学差异。在HFD中补充不同的混合坚果或鞣花酸的小鼠的体重增加率与LFD组没有显著差异。我们发现高营养密度混合坚果比其他两种混合坚果更能轻微缓解高脂饮食诱导的体重增加和肥胖,即使这种差异没有统计学意义。此外,只补充单一核桃促进了小鼠体重增加(p<0.05)
表5高营养密度混合坚果对小鼠体重变化的影响
注:结果为平均值±标准误,*与LFD比较差异显著(p<0.05);**与LFD比较差异显著(p<0.01)
4.2高富含高营养密度混合坚果对小鼠肝脏和脂肪的形态影响
如图1所示,油红O染色的肝脏切片中HFD组的小鼠有明显的肝脏脂肪变性,在三种混合坚果组和EA组中明显减轻(图1A)。此外,HFD组的肝细胞脂滴率明显高于LFD组(P<0.001),高营养密度混合坚果治疗后,肝细胞脂滴率降低(P<0.001,图1D)。H&E染色的肝脏切片显示了类似的形态学变化,这体现在HFD组明显的炎性细胞浸润和脂质空泡化(图1B)。HFD组小鼠的脂肪细胞面积明显大于LFD组(P<0.001),经鞣花酸和混合坚果处理后,特别是高营养密度混合坚果,脂肪细胞面积明显减少(P<0.001,图1C和E)。说明高营养密度混合坚果能显著缓解高脂肪饮食引起的肝脂肪累积。但是,只补充核桃反而增加了肝脏和脂肪组织脂质沉积(P<0.01)。
4.3高富含高营养密度混合坚果对小鼠脂代谢的影响
如图2所示,与LFD组相比,饲喂高脂肪饮食的小鼠血浆中的总胆固醇、甘油三酯和低密度脂蛋白胆固醇含量更高(P<0.05)。饮食添加混合坚果能有效地缓解高脂肪饮食下胆固醇、甘油三酯和高密度脂蛋白胆固醇的升高。三种混合坚果中,尤其是高营养密度混合坚果,能显著调节血脂代谢,降低总胆固醇水平。此外,仅补充核桃无改善血症作用,反而恶化了血浆低密度脂蛋白胆固醇升高(p<0.01)。
4.4高富含高营养密度混合坚果对小鼠糖代谢的影响
如图3所示,与LFD组相比,HFD组的小鼠在灌服葡萄糖后30min和60min的血糖水平升高(P<0.01,图2A),曲线下面积更大(P<0.001,图2B)。三种混合坚果和鞣花酸处理明显改善了HFD引起的血糖升高(P<0.05)。高营养密度混合坚果处理不仅能显著降低30min和60min的血糖水平(P<0.01,图2A),还能降低曲线下面积(P<0.001,图2B)。HFD组胰岛素抵抗指数的增加(P<0.001,图2C),表现出明显的胰岛素抵抗,而高营养密度混合坚果的治疗明显改善了这一点(P<0.05)。说明高营养密度混合坚果能够调节葡萄糖代谢,使血糖维持稳定,显著改善高脂肪饮食诱导的胰岛素抵抗状态。此外,仅补充核桃虽然不会显著降低胰岛素抵抗指数,但却升高了血糖曲线下面积(p<0.01)。
以上所述实施例仅表达了本发明的优选实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形、改进及替代,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (3)
1.一种高营养密度指数混合坚果组合物,其特征在于:包括混合坚果和混合果干;所述混合坚果和混合果干的比例在线性规划计算中单位热量的总富含营养素食物指数最大;
按重量份计算,所述混合坚果为:核桃41.5%、杏仁9%、腰果11.5%、榛子7%;所述混合果干为:黑加仑11%、蓝莓9%、蔓越莓11%;
所述总富含营养素指数的计算公式为:
TNRF=NRF10.3×X+NRF8.3×Y,
其中NRF10.3为所述混合坚果富含营养素指数,
NRF8.3为所述混合果干的富含营养素指数,
X和Y分别为所述混合坚果和所述混合果干的质量占比,X+Y=1;
所述富含营养素指数的计算公式为:
NRFn.3=NR-LIM,
其中,/>,
nutrient i 是第i种鼓励营养素的含量,DVi 是该营养素的每日需要量参考值,MRVi是第i种限制营养素的最大推荐值;
所述混合坚果的10种鼓励营养素为:维生素A、维生素B2、维生素E、铁、镁、锌、鞣花酸、亚麻酸、精氨酸和酪氨酸;所述混合坚果的3种限制营养素为:支链氨基酸、蛋氨酸和饱和脂肪酸;
所述混合果干的8种鼓励营养素为:维生素A、维生素B2、维生素C、镁、鞣花酸、花青素、精氨酸和酪氨酸;所述混合果干的3种限制营养素为:支链氨基酸、蛋氨酸和果糖。
2.一种权利要求1所述高营养密度指数混合坚果组合物的应用,其特征在于:将所述高营养密度指数混合坚果组合物应用于缓解高胆固醇血症或胰岛素抵抗的食品的制备中。
3.根据权利要求2所述的一种高营养密度指数混合坚果组合物的应用,其特征在于:所述食品中还包含常规辅料。
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