CN112155205A - 一种降血压组合物及其制备方法与应用 - Google Patents
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Abstract
本发明涉及一种降血压组合物及其制备方法与应用。所述的降血压组合物,包括如下重量份的原料:腔昆布提取物5~15份、小球藻粉20~30份、苦荞和鹰嘴豆膨化粉28~45份、车前子多糖12~20份、发酵果蔬粉20~30份。本发明还提供上述组合物的制备方法。本发明提供的降血压组合物综合了以上6种成分的功效,合理配伍,安全有效,各种成分通过不同途径进行协同作用,使血管松弛并扩张、减少对血管收缩物质的敏感性、增加短链脂肪酸含量及调节肠道菌群,降压效果更显著。
Description
技术领域
本发明涉及一种降血压组合物及其制备方法与应用,属于功能食品领域。
背景技术
高血压(hypertension)是一种以动脉收缩压或舒张压升高为特征的慢性病,可导致心、脑、肾、血管、眼部结构和功能的改变和损害,引起多种并发症,已成为心血管疾病、心肌梗死、中风等疾病发生的主要因素,目前全球因患高血压死亡人数每年超过700万,也是人类总死亡率的第一危险因素,我国已成为全球高血压危害最严重的国家之一。根据《中国心血管病报告2017(概要)》,中国心血管病患病率及死亡率仍处于上升阶段,心血管病死亡占居民疾病死亡构成40%以上,今后10年心血管病患病人数仍将快速增长。目前心血管病现患人数约2.9亿,其中高血压患者约有2.7亿。高血压导致了70%的脑中风、50%的急性心肌梗死、50%的房颤,还有20%~30%的肾功能不全,因此国家卫建委将高血压作为基层防控最重要的两个抓手之一。
高血压除了与先天的遗传基因相关外,还与后天影响肠道微生态系统的诸多因素有关,其中血管紧张素转换酶(Angiotensin Converting Enzyme,ACE)的高活性被认为是主要的原因之一,第二基因库的肠道微生态系统失衡会导致多个系统的疾病发生。研究表明,原发性高血压病患者氨基酸和脂肪酸代谢紊乱伴随有肠道菌群改变,表现出拟杆菌门菌/厚壁菌门菌比例降低,这种改变也会促进肥胖的发生,从而增加了高血压病的发病风险。同一时期有报道,高血压小鼠肠道中厚壁菌与拟杆菌较正常小鼠明显增多,而产丁酸的细菌却减少了。另有研究表明高血压病患者的肠道菌群数量与同龄正常人群不同,发现直肠真杆菌数量较正常人增多,而多形拟杆菌、双歧杆菌数量较正常人减少。
中国专利文献CN109156757A公开了一种降血压海带提取物,是由发酵的海带经过超纯水提取、干燥、超微粉碎后,获得,制备方法简单、降血压活性强,在1mg/mL的浓度下,对血管紧张素转化酶的抑制率为93.08~93.27%。然而该专利仅仅是提供了一种降压的初级产品,没有发挥出其全部潜力。
传统治疗高血压的方法是药物维持,现在临床常用的降血压药物有卡托普利、苯那普利、氯沙坦、缬沙坦、依那普利和赖诺普利等,它们是通过抑制血管紧张素转换酶(ACE)起到降血压的作用,但它们都是化学合成药,长期服用后会对身体产生副作用,如咳嗽,味觉障碍,高钾血症和皮疹等,所以需要开发天然的、安全无副作用的降压产品。
发明内容
针对现有技术的不足,尤其是目前的降血压产品副作用明显的缺陷,本发明提供一种降血压组合物及其制备方法与应用,本发明的降血压组合物可以使血管松弛并扩张、减少对血管收缩物质的敏感性、调节肠道菌群以及增加短链脂肪酸含量,协同达到控制血压的效果,并且安全无副作用。
本发明的技术方案如下:
一种降血压组合物,包括如下重量份的原料:腔昆布提取物5~15份、小球藻粉20~30 份、苦荞和鹰嘴豆膨化粉28~45份、车前子多糖12~20份、发酵果蔬粉20~30份。
根据本发明优选的,所述降血压组合物,包括如下重量份的原料:腔昆布提取物7~13 份、小球藻粉22~27份、苦荞和鹰嘴豆膨化粉31~40份、车前子多糖14~18份、发酵果蔬粉 23~27份。
根据本发明优选的,所述降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞和鹰嘴豆膨化粉35份、车前子多糖16份、发酵果蔬粉25份。
根据本发明优选的,所述腔昆布提取物规格为10:1。
根据本发明优选的,所述小球藻粉纯度为99%。
根据本发明优选的,所述苦荞和鹰嘴豆膨化粉是按照如下方法制备的:苦荞和鹰嘴豆粉碎至60~80目,将粉碎后的苦荞粉和鹰嘴豆粉按质量比为(1.3~3.8):1混合,搅拌20~30min,然后加入纯化水,得到含水量为6~11%的混合物,接着采用螺杆挤压膨化机对混合物进行膨化,将膨化后的混合物置于70~80℃烘箱中烘干10~50min,使其水分含量为2~5%,经二次粉碎和微波灭菌后完成苦荞和鹰嘴豆的膨化处理。
根据本发明优选的,所述二次粉碎的粒径为80~100目,所述灭菌的条件为:温度65~70℃,时间3~8min。
根据本发明优选的,所述的发酵果蔬粉,包括如下重量份的原料:黑树莓35~44份,蓝莓25~35份,明日叶15~24份、奇数琼胶低聚糖10~16份、低聚果糖7~14份、乳酸菌2~4 份,双歧杆菌2~4份、富硒酵母1~2份。
根据本发明优选的,所述的发酵果蔬粉是按照如下方法制备的:将黑树莓、蓝莓,明日叶按配比混合打浆,加入奇数琼胶低聚糖和低聚果糖,搅拌均匀;然后按配比接种乳酸菌、双歧杆菌和富硒酵母,在24~29.8℃下发酵20~26天,发酵结束后进行膜过滤,得澄清液,所得澄清液在95~105℃条件下灭菌10~20min后经过喷雾干燥,得到发酵果蔬粉。
根据本发明优选的,所述的乳酸菌为噬酸乳杆菌LA-G80、保加利亚乳杆菌LB-G40和鼠李唐乳杆菌Lr-G14,按照质量比为1:1:1的比例混合的混合物。
根据本发明优选的,所述的双歧杆菌为乳双歧杆菌BL-G101、两歧双歧杆菌BB-G90、短双歧杆菌BB-G95,按照质量比为1:1:1的比例混合的混合物。
上述降血压组合物的制备方法,包括步骤如下:
(1)将腔昆布提取物、小球藻粉、苦荞和鹰嘴豆膨化粉、车前子多糖、发酵果蔬粉各原料过80目筛,按比例混合均匀,得混合物;
(2)向混合物中加入酒精,制得软材;然后过20目筛进行制粒,制粒后于50~60℃烘干2~3h,再经整粒,筛粉后得到降血压组合物。
根据本发明优选的,步骤(2)中,所述的酒精浓度为40~60%,酒精的加入量为混合物质量的4~12%。
本发明所述的降血压的组合物在制备具有降血压功效的食品、药品或保健品中的应用。
本发明中所述的喷雾干燥按现有技术进行处理。
本发明的技术特点在于:
本发明采用的腔昆布提取物,通过肾素-血管紧张素系统与激肽释放酶-激肽系统和内皮依赖性血管收缩因子和舒张因子系统调节血压,达到降血压功效。腔昆布提取物对血管紧张素转化酶(ACE)有抑制效果,其提取物中的二鹅掌菜酚能诱导细胞产生NO,两个系统协同作用,达到降血压的效果。
本发明采用的小球藻(Chlorella)是一类普生性单细胞绿藻,属于绿藻门、小球藻属,营养丰富,含有降血压肽,通过肾素-血管紧张素系统与激肽释放酶-激肽系统调节血压,达到降血压的功效。研究表明,从小球藻废弃蛋白中分离到具有降压活性的11肽,与血管紧张素转化酶(ACE)的结合模式是非竞争性的;相类似的,用碱性蛋白酶水解椭圆小球藻得到的产物具有最高的ACE抑制活性,竞争性抑制ACE,并且通过饲喂致病原发性高血压小鼠发现其可以显著降低高血压症状。
本发明选择的苦荞麦,寥科属,双子叶植物,属于药食兼用作物。苦荞麦籽实中生物活性物质含量丰富,其中的槲皮素和桑色素等具有改善机体血管平滑肌的收缩和舒张的功能,芦丁具有扩张血管的作用,从而起到调节血压的功效。苦荞中膳食纤维含量丰富,而高纤维膳食能够增加更多的产生乙酸和丁酸的细菌,进而通过短链脂肪酸改善高血压。
本发明选择的鹰嘴豆,别名桃豆、鸡豌豆,豆科鹰嘴豆属,豆科蝶形花亚科典型代表植物。鹰嘴豆中所含的营养成分非常丰富,无论是从种类,还是数量上,都大大超过其他豆类。此外,鹰嘴豆富含可溶性膳食纤维,能促进人体中的肠道微生物生成丁酸,短链脂肪酸通过 GPR43和GPR109A信号通路改善血管的张力,从而降低了血压。
本发明选择的车前子多糖是车前草的种皮细胞壁中所含的黏胶,有良好的亲水性,具有调节肠道菌群的作用。车前子多糖对高血压患者肠道菌群微生态具有纠正作用,并且对肠道双歧杆菌和多形拟杆菌有促生长的作用,对直肠真杆菌具有抑制作用。
本发明中的发酵果蔬粉含有丰富的降血压活性物质,通过使血管松弛并扩张,达到调节血压的功效;同时硒通过调控Ca2+的作用,减少对血管收缩物质的敏感性,从而降低血压;此外,能调节肠道菌群,促进有益菌生长,抑制有害菌繁殖,进而调节血压。
本发明能够带来如下有益效果:
1、本发明提供的降血压组合物腔昆布提取物与小球藻粉复配通过肾素-血管紧张素系统与激肽释放酶-激肽系统和内皮依赖性血管收缩因子和舒张因子系统协同调节血压;苦荞和鹰嘴豆协同增加肠道中短链链脂肪酸含量,改善血管张力,达到降血压的效果;车前子多糖与发酵果蔬粉协同调节肠道菌群,增加有益菌相对丰度,抑制有害菌繁殖,实现降血压功效。本发明提供的降血压组合物综合了以上6种成分的功效,合理配伍,安全有效,各种成分通过不同途径进行协同作用,使血管松弛并扩张、减少对血管收缩物质的敏感性、增加短链脂肪酸含量及调节肠道菌群,降压效果更显著。
2、本发明的苦荞和鹰嘴豆膨化粉采用混合膨化处理,膨化处理可以有效去除了鹰嘴豆中的毒素和难消化物质,很好的保留了苦荞和鹰嘴豆中的膳食纤维含量,提高可溶性膳食纤维含量,同时显著提高了苦荞和鹰嘴豆的消化率及吸收利用率。
3、本发明的发酵果蔬粉,通过添加益生菌和富硒酵母进行发酵得到,碳源采用奇数琼胶低聚糖和低聚果糖,奇数琼胶低聚糖能增加Alistipes和Akk菌等有益菌和减少螺杆菌的相对丰度;采用发酵方式能更好的保留果蔬粉的生物活性物质,同时添加富硒酵母增加了硒的含量,与果蔬的生物活性物质协同降血压。
附图说明
图1为实验例3对目标菌群的分析图。
具体实施方式
下面结合具体实施例对本发明进行详细说明,以下实施例仅为方便本领域技术人员理解本发明技术方案,实现或使用本发明所做的说明,并不以此限制本发明的保护范围。
腔昆布提取物,陕西斯诺特生物技术有限公司有售;小球藻粉,西安瑞尔丽生物工程有限公司有售;苦荞,宣威市民望荞业发展有限公司有售;鹰嘴豆,罗庄区亿鑫粮行有售;车前子多糖,陕西新天域生物科技有限公司有售;黑树莓、蓝莓、明日叶,各大超市有售;乳酸菌和双歧杆菌,润盈生物工程(上海)有限公司有售;富硒酵母,安琪酵母股份有限公司有售;奇数琼胶低聚糖,青岛博智汇力生物科技有限公司有售;低聚果糖,保龄宝生物股份有限公司有售。
实施例1
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞和鹰嘴豆膨化粉35份、车前子多糖16份、发酵果蔬粉25份。
所述腔昆布提取物规格为10:1,小球藻粉的纯度为99%。
所述的发酵果蔬粉,按重量份计,包括如下原料:黑树莓40份,蓝莓30份,明日叶20份、奇数琼胶低聚糖13份、低聚果糖10份、乳酸菌3份,双歧杆菌3份、富硒酵母1.5份。
所述苦荞和鹰嘴豆膨化粉是按照如下方法制备的:苦荞和鹰嘴豆粉碎至60目,将粉碎后的苦荞粉和鹰嘴豆粉按质量比为2:1混合,搅拌20min,然后加入纯化水,得到含水量为6%的混合物,接着采用螺杆挤压膨化机对混合物进行膨化,将膨化后的混合物置于70℃烘箱中烘干20min,使其水分含量为2%,经二次粉碎至80目,在65℃下微波灭菌5min后完成苦荞和鹰嘴豆的膨化处理。
所述的发酵果蔬粉是按照如下方法制备的:将黑树莓、蓝莓、明日叶按配比混合打浆;加入奇数琼胶低聚糖和低聚果糖,搅拌均匀;然后按配比接种乳酸菌、双歧杆菌和富硒酵母,在24℃下发酵20天,发酵结束后采用孔径100nm进行膜过滤,得澄清液,所得澄清液在95℃条件下灭菌10min后经过喷雾干燥,得到发酵果蔬粉。
所述的乳酸菌为噬酸乳杆菌LA-G80、保加利亚乳杆菌LB-G40和鼠李唐乳杆菌Lr-G14,按照质量比为1:1:1的比例混合的混合物。所述的双歧杆菌为乳双歧杆菌BL-G101、两歧双歧杆菌BB-G90、短双歧杆菌BB-G95,按照质量比为1:1:1的比例混合的混合物。
实施例2
一种降血压组合物,包括如下重量份的原料:腔昆布提取物15份、小球藻粉20份、苦荞和鹰嘴豆膨化粉32份、车前子多糖20份、发酵果蔬粉20份。
所述的发酵果蔬粉,按重量份计,包括如下原料:黑树莓44份,蓝莓35份,明日叶24份、奇数琼胶低聚糖16份、低聚果糖14份、乳酸菌4份,双歧杆菌4份、富硒酵母2份。
所述腔昆布提取物规格为10:1,小球藻粉的纯度为99%。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的制备过程同实施例1。
实施例3
一种降血压组合物,包括如下重量份的原料:腔昆布提取物5份、小球藻粉30份、苦荞和鹰嘴豆膨化粉38份、车前子多糖12份、发酵果蔬粉30份。
所述的发酵果蔬粉,按重量份计,包括如下原料:黑树莓35份,蓝莓25份,明日叶15份、奇数琼胶低聚糖10份、低聚果糖7份、乳酸菌2份,双歧杆菌2份、富硒酵母1份。
所述腔昆布提取物规格为10:1,小球藻粉的纯度为99%。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的制备过程同实施例1。
实施例4
一种降血压组合物的制备方法,包括步骤如下:
(1)将腔昆布提取物、小球藻粉、苦荞和鹰嘴豆膨化粉、车前子多糖、发酵果蔬粉各原料过80目筛,按比例混合均匀,得混合物;
(2)向混合物中,按混合物质量的10%加入浓度为50%的酒精,制得软材;然后过20目筛进行制粒,制粒后于55℃烘干3h,再经整粒,筛粉后得到降血压组合物。
对比例1
一种降血压组合物,包括如下重量份的原料:腔昆布提取物17份、小球藻粉40份、苦荞和鹰嘴豆膨化粉60份、车前子多糖26份、发酵果蔬粉40份。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例2
一种降血压组合物,包括如下重量份的原料:腔昆布提取物3份、小球藻粉10份、苦荞和鹰嘴豆膨化粉13份、车前子多糖6份、发酵果蔬粉10份。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例3
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞和鹰嘴豆膨化粉35份、车前子多糖16份。
苦荞和鹰嘴豆的膨化过程同实施例1。
对比例4
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞和鹰嘴豆膨化粉35份、发酵果蔬粉25份。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例5
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞膨化粉25份、车前子多糖16份、发酵果蔬粉25份。
苦荞的膨化过程如下:苦荞粉碎至60目,向粉碎后的苦荞粉加入纯化水,使其含水量为 6%,然后采用螺杆挤压膨化机进行膨化,将膨化后的苦荞粉置于70℃烘箱中烘干20min,使其水分含量为2%,经二次粉碎至80目,在65℃下微波灭菌5min后完成苦荞的膨化处理。
发酵果蔬粉的原料组成和制备过程同实施例1。
对比例6
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、鹰嘴豆膨化粉10份、车前子多糖16份、发酵果蔬粉25份。
鹰嘴豆的膨化过程同实施例5,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例7
一种降血压组合物,包括如下重量份的原料:腔昆布提取物10份、苦荞和鹰嘴豆膨化粉 35份、车前子多糖16份、发酵果蔬粉25份。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例8
一种降血压组合物,包括如下重量份的原料:小球藻粉25份、苦荞和鹰嘴豆膨化粉35 份、车前子多糖16份、发酵果蔬粉25份。
苦荞和鹰嘴豆的膨化过程同实施例1,发酵果蔬粉的原料组成和制备过程同实施例1。
对比例9
按照中国专利文献CN109156757A获得海带提取物。
实验例1
根据保健食品检验与评价技术规范(2003版)第九章辅助降血压功能检验方法,本发明组合物的降血压功效以遗传性高血压模型法进行评价。
试验对象:大鼠试验中选用自发性高血压大鼠(SHR),雄性,清洁级,周龄10~12,体重180~220g;SPF级雄性Wistar大鼠,体重180~220g,血压124.10±10.63mmHg。
试验方法:
试验前一周对受试动物采用尾脉搏进行多次血压测量,使其适应测压环境。自发高血压大鼠(SHR)70只,根据基础血压值随机平均分为7组。Wistar大鼠10只,为普通动物对照组。 SHR大鼠设模型对照组和实验组1~6,实验组1~3组分别给予实施例1~3的组合物,实验组 4~5组分别给予对比例1~2的组合物,实验组6给予对比例9的海带提取物,人体推荐量为每天2.8g/60kg.bw(仅主药量),按中剂量组466.6mg/kg.bw(相当于人体推荐摄入量的10倍),以纯化水为溶剂配至所需浓度,每只大鼠按1.0mL/100g.bw灌胃,模型对照组灌胃纯化水,普通动物对照组按实施例1高剂量1399.9mg/kg.bw(相当于人体推荐摄入量的30倍),以纯化水为溶剂配至所需浓度,每只大鼠按1.0mL/100g.bw灌胃,持续30d。
采用智能无创血压计,测定试验大鼠动脉收缩压。测压前,将大鼠置于大鼠网保温套中,并根据每只动物体型用鼠袋将其固定,在40℃下恒温预热10min左右,促进尾动脉有效扩张,血流通畅;然后将大鼠加压感应器套入鼠尾根部。试验期间、用药前及用药后每周测量血压、心率和体重各1次。
使用SPSS统计软件进行数据处理,t检验进行各组间显著性差异分析,P<0.05为差异显著,P<0.01为差异非常显著。所有检测指标的数据均以均数±标准差表示,测试结果见表 1。
表1:实验组1~6、模型对照组及普通动物组对试验大鼠体重、血压、心率的影响
序号 | 组别 | 体重/g | 血压/mmHg | 心率/次/分 |
1 | 普通动物组 | 345.64±28.16 | 123.28±14.50 | 380.2±23.6 |
2 | 模型对照组 | 341.37±27.94 | 196.04±15.91 | 390.5±24.3 |
3 | 实验组1 | 320.74±28.81 | 162.49±30.36** | 375.4±18.56 |
4 | 实验组2 | 325.99±31.04 | 172.67±22.26* | 382.9±20.9 |
5 | 实验组3 | 328.20±35.38 | 176.97±21.99* | 378.3±23.3 |
6 | 实验组4 | 332.93±29.26 | 190.19±18.94 | 383.9±21.9 |
7 | 实验组5 | 336.14±36.02 | 193.18±21.98 | 385.9±20.8 |
8 | 实验组6 | 333.48±31.19 | 179.36±17.60* | 382.2±21.4 |
注:*表示与模型对照组比较,*P<0.05,**P<0.01。
试验过程中各组动物生长良好,活动正常。由表1可知,实验组1~6与模型对照组体重均无显著性差异(P>0.05);普通动物组血压值与试验前无显著性差异,实验组1血压值与模型对照组相比,具有极显著差异(P<0.01),实验组2~3中血压值对于模型对照组P<0.05,表明具有显著差异性,实验组4~5中血压值对于模型对照组P>0.05,表明无显著差异性;实验组6中血压值对于模型对照组P<0.05,具有差异性,试验对各组大鼠心率均无影响,各组间心率也无显著性差异。
以上实验结果表明,本发明实施例1~3的组合物相较于对比例1~2的组合物,在调整各成分用量后,对大鼠降血压的作用明显增强,相较于对比例9的海带提取物本发明实施例1 的组合物对大鼠降血压的作用更加显著。
实验例2
根据实验例1的实验结果,实验样品选取实施例1中的组合物,人体推荐量为每天2.8g/60kg.bw(仅主药量),实验设低、中、高三个剂量组,分别为233.3mg/kg.bw、 466.6mg/kg.bw、1399.9mg/kg.bw,低、中、高三个组的剂量分别相当于人体推荐摄入量的5 倍、10倍、30倍,依然以大鼠实验进行评价,将对比例3~8的组合物按高剂量组灌入。实验方法与实验例1中相同,试验组SHR大鼠随机平分10组,分别为模型对照组和实验组1~9,实验组1~3分别以实施例1低中高三个剂量,实验组4~9以对比例3~8中组合物的配比,以纯化水为溶剂配至所需浓度,每只大鼠按1.0mL/100g.bw灌胃,模型对照组灌胃纯化水,普通动物对照组按实施例1高剂量1399.9mg/kg.bw(相当于人体推荐摄入量的30倍),以纯化水为溶剂配至所需浓度,每只大鼠按1.0mL/100g.bw灌胃。大鼠指标的测定和数据处理同实验例1,各组实验指标如表2所示:
表2:实验组1~9、模型对照组及普通动物组对试验大鼠体重、血压、心率的影响
序号 | 组别 | 体重/g | 血压/mmHg | 心率/次/分 |
1 | 普通动物组 | 341.39±25.99 | 124.40±15.30 | 376.7±19.4 |
2 | 模型对照组 | 338.68±19.51 | 197.39±19.96 | 385.4±22.1 |
3 | 实验组1 | 329.07±25.36 | 173.72±24.14* | 378.3±20.1 |
4 | 实验组2 | 324.17±25.28 | 163.88±26.64** | 373.9±18.1 |
5 | 实验组3 | 321.11±23.71 | 157.01±16.72** | 370.2±18.5 |
6 | 实验组4 | 331.13±25.49 | 185.23±23.29 | 380.1±18.1 |
7 | 实验组5 | 335.22±25.65 | 186.38±19.74 | 384.8±19.1 |
8 | 实验组6 | 333.29±27.98 | 188.46±18.80 | 378.6±22.8 |
9 | 实验组7 | 337.27±20.65 | 190.37±21.63 | 383.3±23.2 |
10 | 实验组8 | 334.83±28.10 | 192.16±19.52 | 382.4±19.7 |
11 | 实验组9 | 336.15±25.17 | 193.26±22.11 | 386.4±20.6 |
注:*表示与模型对照组比较,*P<0.05,**P<0.01。
试验过程中各组动物生长良好,活动正常。由表2可知,实验组1~9与模型对照组体重 P>0.05,无显著差异性;普通动物组血压值与试验前无显著性差异,试验组1血压值与模型对照组相比,具有显著差异性(P<0.05),实验组2~3中血压值对于模型对照组P<0.01,表明具有极显著差异性,实验组4~9中血压值对于模型对照组P>0.05,表明无显著差异性;试验对各组大鼠心率均无影响,各组间心率也无显著性差异。
以上数据表明,本发明提供的降血压组合物中腔昆布提取物、小球藻粉、苦荞、鹰嘴豆、车前子多糖及发酵果蔬粉产生协同,可以使血管松弛并扩张、减少对血管收缩物质的敏感性、增加短链脂肪酸含量和调节肠道菌群,从而达到降血压功效。
实验例3对肠道菌群的调节作用
大鼠粪便的收集:在实验例1中大鼠饲养周期末,将各笼大鼠的垫料去除后,分别收集各笼大鼠排出的新鲜粪便,称取粪便2g于2mL的已做好标记的无酶冷存管,快速置于-80℃冰箱保存。
荧光定量PCR法测定目标菌群拷贝数:各目标菌群引物序列参考已有文献中的序列,如表3所示,各目标菌群的拷贝数根据各样品的Ct值和相应标准曲线的回归方程准确计算出样品菌群的含量,换算成每毫升样品中基因的拷贝数(copies/μL),结果如图1所示。
表3:各目标菌群引物序列
双歧杆菌在肠道菌群中占有优势地位,其产生的磷壁酸可使其定植于肠黏膜上皮,与肠道中的其他厌氧细菌合作形成保护屏障,同时,能降低肠道pH值,抑制有害菌的繁殖,防止病原菌的定植和侵袭,对稳定肠道菌群结构方面起着重要作用。多形拟杆菌是拟杆菌门的代表,能降解人体肠道中大部分的植物多糖、胆汁酸并参与类固醇代谢,多形拟杆菌数量的变化可能会影响固醇类的正常代谢,从而增加高血压的发病风险。直肠真杆菌为条件致病菌,是厚壁菌门的代表菌,高血压病患者肠道中该菌较正常人明显增加。
本发明中,模型对照组与普通动物组比较,双歧杆菌、多形拟杆菌拷贝数显著减少(P< 0.05),直肠真杆菌拷贝数显著增加(P<0.05);实验组1~3双歧杆菌、多形拟杆菌拷贝数对比模型对照组显著增加(P<0.05),具有显著差异性,直肠真杆菌拷贝数对比模型对照组显著减少(P<0.05),具有显著差异性;实验组4~5中双歧杆菌、多形拟杆菌及直肠真杆菌拷贝数与模型对照组的P>0.05,差异不显著;本实验例对目标菌群的分析图如图1所示。
以上数据表明,实验组1~3的降血压组合物可以显著增加大鼠肠道中双歧杆菌、多形拟杆菌数量,减少直肠真杆菌数量,进而通过调节肠道菌群达到降血压的功效。
实验例4ACE抑制活性的测定
测定方法如下:
ACE溶液:将2.5U ACE溶于冷的1mol/L pH 8.3的硼酸盐缓冲液(含0.3mol/LNaCl)10mL 配制成250mU/mL ACE溶液,分装,储存于-20℃,备用。
HHL(马尿酰组氨酰亮氨酸)溶液:取HHL适量,以0.1mol/L pH 8.3的硼酸盐缓冲液(含 0.3mol/L NaCl)配制成质量浓度为0.3%的HHL溶液,备用。
马尿酸标准溶液:准确称取马尿酸标准样品,加超纯水溶解配制成浓度为50μg/mL马尿酸标准溶液,备用。
抑制剂溶液:将实施例1中的降血压组合物配置成1mg/mL的溶液,对比例9中的海带提取物配置成1mg/mL的溶液。
反应体系环境为pH 8.3,0.1mol/L硼酸盐缓冲液(含有0.3mol/L NaCl)。于1.5mL离心管中,取50μL 0.3%HHL,加入20μL抑制剂溶液,混合均匀,37℃±0.5℃恒温水浴中预热5min,然后加入20μL 250mU/mL ACE,充分混匀。37℃恒温水浴保温60min后,加入10μL0.2mol/L HCl中止反应。同时用20μL水代替样品作为空白对照组。反应液离心后用高相液相色谱仪检测Hip生成量。
色谱条件:Ghall 12S05-2546 C18柱(250mm×4.6mm);乙腈:水(含0.05%三氟乙酸)= 25:75,等梯度洗脱;流速0.5mL/min;检测波长228nm;进样量10μL。
ACE抑制率按下式进行计算:
式中:
A样品——加入抑制剂溶液中马尿酸的峰面积;
A对照——空白对照组中马尿酸的峰面积。
本实验例中,实施例1降血压组合物对血管紧张素转化酶的抑制率为94.2%,对比例9 海带提取物对血管紧张素转化酶的抑制率93.1%,说明本发明通过肾素-血管紧张素系统与激肽释放酶-激肽系统改善血压优于海带提取物的效果,同时本发明降血压组合物综合了6 种成分,合理配伍,各种成分通过不同途径进行协同作用,相较于海带提取物降压效果更显著。
以上所述仅为本申请的实施例而已,并不用于限制本申请。对于本领域技术人员来说,本申请可以有各种更改和变化。凡在本申请的精神和原理之内所作的任何修改、等同替换、改进等,均应包含在本申请的权利要求范围之内。
Claims (10)
1.一种降血压组合物,包括如下重量份的原料:腔昆布提取物5~15份、小球藻粉20~30份、苦荞和鹰嘴豆膨化粉28~45份、车前子多糖12~20份、发酵果蔬粉20~30份。
2.如权利要求1所述的降血压组合物,其特征在于,所述降血压组合物,包括如下重量份的原料:腔昆布提取物7~13份、小球藻粉22~27份、苦荞和鹰嘴豆膨化粉31~40份、车前子多糖14~18份、发酵果蔬粉23~27份。
3.如权利要求1所述的降血压组合物,其特征在于,所述降血压组合物,包括如下重量份的原料:腔昆布提取物10份、小球藻粉25份、苦荞和鹰嘴豆膨化粉35份、车前子多糖16份、发酵果蔬粉25份。
4.如权利要求1所述的降血压组合物,其特征在于,所述苦荞和鹰嘴豆膨化粉是按照如下方法制备的:苦荞和鹰嘴豆粉碎至60~80目,将粉碎后的苦荞粉和鹰嘴豆粉按质量比为(1.3~3.8):1混合,搅拌20~30min,然后加入纯化水,得到含水量为6~11%的混合物,接着采用螺杆挤压膨化机对混合物进行膨化,将膨化后的混合物置于70~80℃烘箱中烘干10~50min,使其水分含量为2~5%,经二次粉碎和微波灭菌后完成苦荞和鹰嘴豆的膨化处理。
5.如权利要求1所述的降血压组合物,其特征在于,所述的发酵果蔬粉,包括如下重量份的原料:黑树莓35~44份,蓝莓25~35份,明日叶15~24份、奇数琼胶低聚糖10~16份、低聚果糖7~14份、乳酸菌2~4份,双歧杆菌2~4份、富硒酵母1~2份。
6.如权利要求1所述的降血压组合物,其特征在于,所述的发酵果蔬粉是按照如下方法制备的:将黑树莓、蓝莓,明日叶按配比混合打浆;加入奇数琼胶低聚糖和低聚果糖,搅拌均匀;然后按配比接种乳酸菌、双歧杆菌和富硒酵母,在24~29.8℃下发酵20~26天,发酵结束后进行膜过滤,得澄清液,所得澄清液在95~105℃条件下灭菌10~20min后经过喷雾干燥,得到发酵果蔬粉。
7.如权利要求1所述的降血压组合物,其特征在于,所述的乳酸菌为噬酸乳杆菌LA-G80、保加利亚乳杆菌LB-G40和鼠李唐乳杆菌Lr-G14,按照质量比为1:1:1的比例混合的混合物;所述的双歧杆菌为乳双歧杆菌BL-G101、两歧双歧杆菌BB-G90、短双歧杆菌BB-G95,按照质量比为1:1:1的比例混合的混合物。
8.权利要求1所述的降血压组合物的制备方法,包括步骤如下:
(1)将腔昆布提取物、小球藻粉、苦荞和鹰嘴豆膨化粉、车前子多糖、发酵果蔬粉各原料过80目筛,按比例混合均匀,得混合物;
(2)向混合物中加入酒精,制得软材;然后过20目筛进行制粒,制粒后于50~60℃烘干2~3h,再经整粒,筛粉后得到降血压组合物。
9.如权利要求8所述的制备方法,步骤(2)中,所述的酒精浓度为40~60%,酒精的加入量为混合物质量的4~12%。
10.权利要求1-7所述的降血压组合物在制备具有降血压功效的食品、药品或保健品中的应用。
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