CN111227233B - 丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用 - Google Patents
丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用 Download PDFInfo
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- Medicines Containing Plant Substances (AREA)
Abstract
本发明涉及食品营养技术领域,具体涉及丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用。为了达到上述目的,本发明提供了如下技术方案:一种丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用。本发明中,证明丝瓜粉通过改善高支链氨基酸血症抑制肥胖发生。本发明中,丝瓜粉的制备方法包括:丝瓜清洗切片、低温冷冻干燥、超微粉碎。此方法可以最大程度保留丝瓜的营养成分。由结果可知,丝瓜粉能够上调肥胖小鼠脂肪组织和肝脏中支链氨基酸分解代谢酶基因转录水平的表达,促进体内支链氨基酸分解代谢,改善高支链氨基酸血症,抑制肥胖及胰岛素抵抗,可作为通过改善高支链氨基酸血症抑制肥胖发生的膳食营养成分。
Description
技术领域
本发明涉及食品营养技术领域,具体涉及丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用。许多临床研究表明高支链氨基酸血症可以作为预防或治疗肥胖及其代谢综合症的诊断指标。经动物实验证明,低温冷冻干燥制备的丝瓜粉可以通过上调支链氨基酸分解代谢酶的表达促进体内多余支链氨基酸的分解代谢,改善高支链氨基酸血症,进而缓解肥胖等代谢疾病,可作为膳食添加剂或者饮食结构成分用于改善支链氨基酸代谢紊乱抑制肥胖发生。
背景技术
流行病学表明肥胖的发病率在发达国家和发展中国家迅速攀升,并且患病呈现年轻化的态势,肥胖引起的并发症多种多样。现代饮食习惯(高脂肪、低膳食纤维)引起的肥胖会导致人体生理稳态失衡(脂代谢,糖代谢,氨基酸代谢),同时伴随高血脂、高血糖、二型糖尿病、心脑血管疾病和某些癌症等疾病的发生。
许多研究证明机体内支链氨基酸水平的高低可用于诊断肥胖及其代谢疾病的发病病程。支链氨基酸(BCAAs,branched-chain amino acids)包括缬氨酸(Val)、亮氨酸(Leu)、异亮氨酸(Ile),是人体重要的营养必需氨基酸。食物是人体内BCAA的主要来源,正常生理状况下机体内BCAA分解代谢保持动态平衡。BCAAs分解代谢主要发生在脂肪组织和肝脏组织的线粒体中,第一步是由支链氨基酸转氨酶(BCATm,mitochondrial branched-chain amino acid)催化生成支链α酮酸,第二步是支链α酮酸脱氢酶激酶(BCKDK,branched-chainα-keto acid dehydrogenase kinase)复合物会进一步氧化脱羧,进入三羧酸循环生成NADH、CO2等终产物。大量研究表明长期高脂饮食摄入导致体内BCAA代谢紊乱,具体表现是BCAA分解代谢酶表达降低,血液中支链氨基酸过度积累,进而诱发或加重肥胖及胰岛素抵抗等病症。近年来,人们逐渐关注采用饮食疗法通过多种途径预防和缓解肥胖及其代谢并发症,寻找有效改善相关疾病的膳食或者饮食组成有重要意义。
丝瓜又名绵瓜、布瓜和蛮瓜等,为葫芦科丝瓜属一年生攀援草本植物,全球种植广泛。丝瓜药食两用,其鲜嫩肉质果实属瓠果,不仅富含膳食纤维、维生素、多酚、黄酮类、皂苷、三萜类、齐墩果酸、胡萝卜素以及叶绿素等营养元素,还可作为中草药用于清热解毒、咳嗽发烧、呼吸困难和皮下肿胀的病症,是具有良好健康功效的蔬菜食品。本专利中所制备的丝瓜粉经营养成分测定含有51.47%碳水化合物,21.75%蛋白质,0.7%脂肪,5.34%灰分以及20.74%膳食纤维,能量密度为3.04Kal/g,具有低脂低热量的良好特性。而丝瓜果实老熟干燥之后的种子是丝瓜子,又名乌牛子,呈扁平椭圆形,种皮灰黑色或黑色,口味微苦。丝瓜子富含多种天然脂类等营养物质,据报道丝瓜子约含脂肪油24-39%,以亚油酸、棕榈酸、硬脂酸、油酸等为主,具有利水除热,补气益中,通便驱虫的功效。中国专利2012106544160.3提供丝瓜子用于预防缓解与血脂升高相关疾病等应用。中国专利201510577827.3提供丝瓜提取物用于预防和治疗肥胖中的应用。但目前,针对丝瓜通过改善高支链氨基酸血症抑制肥胖发生的应用未见报道。
发明内容
本发明目的在于提供一种丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用。
为了达到上述目的,本发明提供了如下技术方案:
一种丝瓜粉用于改善高支链氨基酸血症抑制肥胖发生的应用。
其中,所述丝瓜粉的制备方法包括如下步骤:
1)丝瓜清洗切片:摘取新鲜丝瓜,清洗干净后切成薄片;
2)低温冷冻干燥:步骤1中的丝瓜薄片在-20℃至-80℃下预冻24h-48h,然后在-60℃至-80℃下低温冷冻干燥48h-72h;
3)超微粉碎:步骤2中经冷冻干燥的丝瓜薄片超微粉碎后过60目筛后得到丝瓜粉。
其中,步骤1中,摘取的新鲜丝瓜清洗干净后切至2-3mm厚度薄片。
其中,步骤2中,步骤1中的丝瓜薄片在-80℃下预冻48h,然后在-80℃下低温冷冻干燥72h。
其中,所述丝瓜粉的制备方法还包括:
4)步骤3制备得到的丝瓜粉,避光-20℃保存。
其中,步骤3中得到的丝瓜粉,使用前用无菌生理盐水混合成0.2g/mL配制丝瓜悬浊液。
与现有技术相比,本发明的有益效果在于:本发明中,证明丝瓜粉通过改善高支链氨基酸血症抑制肥胖发生。
本发明中,丝瓜粉的制备方法包括:丝瓜清洗切片、低温冷冻干燥、超微粉碎。此方法可以最大程度保留丝瓜的营养成分。
由结果可知,丝瓜粉能够上调肥胖小鼠脂肪组织和肝脏中支链氨基酸分解代谢酶基因转录水平的表达,促进体内支链氨基酸分解代谢,改善高支链氨基酸血症,抑制肥胖及胰岛素抵抗,可作为通过改善高支链氨基酸血症抑制肥胖发生的膳食营养成分。
附图说明
图1显示丝瓜粉对高脂饮食小鼠脂肪组织和肝脏组织中支链氨基酸分解代谢酶基因表达的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05);
图2显示丝瓜粉对高脂饮食小鼠血清中支链氨基酸水平的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05);
图3、图4显示丝瓜粉对高脂饮食小鼠体重和脂肪分布的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05);
图5显示丝瓜粉对高脂饮食小鼠脂肪组织脂质堆积的影响;
图6显示丝瓜粉对高脂饮食小鼠肝脏组织脂质堆积的影响。
具体实施方式
下面结合附图对本发明的具体实施方式作进一步说明。
本发明的丝瓜粉的制备方法包括如下步骤:
1、丝瓜清洗切片:摘取新鲜丝瓜,清洗干净后切成薄片;
优选地,步骤1中,摘取的新鲜丝瓜清洗干净后切至2-3mm厚度薄片。
2、低温冷冻干燥:步骤1中的丝瓜薄片在-20℃至-80℃下预冻24h-48h,然后在-60℃至-80℃下低温冷冻干燥48h-72h。
优选地,步骤2中,步骤1中的丝瓜薄片在-80℃下预冻48h,然后在-80℃下低温冷冻干燥72h。
3、超微粉碎:步骤2中经冷冻干燥的丝瓜薄片超微粉碎后过60目筛后得到丝瓜粉。
优选地,本发明的丝瓜粉的制备方法还包括:
4、步骤3制备得到的丝瓜粉,避光-20℃保存。
优选地,步骤3中得到的丝瓜粉,使用前用无菌生理盐水混合成0.2g/mL配制丝瓜悬浊液。
通过此方法制备得到的丝瓜粉,可以最大程度保留丝瓜的营养成分。
丝瓜粉对高脂饮食小鼠脂肪组织和肝脏组织中支链氨基酸分解代谢酶基因表达的影响、丝瓜粉对高脂饮食小鼠血清中支链氨基酸水平的影响、丝瓜粉对高脂饮食小鼠体重和脂肪分布的影响、丝瓜粉对高脂饮食小鼠脂肪组织脂质堆积的影响、丝瓜粉对高脂饮食小鼠肝脏组织脂质堆积的影响:
1、丝瓜干预物制备
摘取新鲜丝瓜,清洗干净后切至2-3mm厚度薄片,-80℃预冻48h并-80℃低温冷冻干燥72h,超微粉碎后过60目筛后得到丝瓜粉,避光-20℃保存。使用前用无菌生理盐水混合成0.2g/mL丝瓜粉悬浊液,灌胃剂量是10μL/g小鼠体重。
2、动物实验
1)30只5周龄健康C57BL/6J雄性小鼠(北京维通利华公司提供),适应性喂养一周后,随机分为3组,5只一笼,每组10只。第一组喂食对照饲料(D12450B,3.85Kcal/g,购自美国Research Diets公司)并灌胃等剂量生理盐水,即对照组;第二组喂食高脂饲料(D12492,5.24Kcal/g,购自美国Research Diets公司)并灌胃等剂量生理盐水,即高脂组;第三组喂食高脂饲料并灌胃0.2g/mL丝瓜粉悬浊液,即丝瓜干预组。实验时间12周,每天上午8:30-9:30灌胃处理,实验期间小鼠对饲料和饮水自由摄取。
2)实验结束,称取各组小鼠体重,收集各组小鼠血清、肝脏组织及附睾脂肪组织,血清样本进行血生化指标总甘油三酯TG、总胆固醇TC、高密度脂蛋白HDL、低密度脂蛋白LDL、空腹血糖Glu、空腹胰岛素INS检测,用ELISA试剂盒检测血清中游离脂肪酸FFA含量。计算各组小鼠胰岛素抵抗指数HOMA-IR(HOMA-IR=(Glu(mmol/L)×INS(mIU/L))/22.5)。部分肝脏和脂肪组织用于病理检验及组织学分析,部分肝脏及脂肪组织用于支链氨基酸分解代谢酶基因转录水平表达分析。
所得结果如下:
图1显示丝瓜粉对高脂饮食小鼠脂肪组织和肝脏组织中支链氨基酸分解代谢酶基因表达的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05)。
图1表明与对照组相比,高脂饮食导致小鼠脂肪组织中BCAA分解代谢关键酶BCATm和BCKDK的mRNA水平表达显著降低大于50%,丝瓜干预组小鼠BCATm和BCKDK的mRNA水平表达可恢复至对照组水平;高脂饮食小鼠肝脏组织中BCATm的mRNA水平表达同样明显降低约50%,丝瓜粉干预组BCATm的mRNA水平表达显著恢复,与对照组基本相同;虽然高脂饮食小鼠肝脏中BCKDK的mRNA水平表达与对照组相比升高接近两倍,但丝瓜干预组小鼠肝脏中BCKDK的表达与对照组和高脂组相比无显著性差异;因此,丝瓜粉干预可明显提高高脂饮食小鼠脂肪组织和肝脏组织中支链氨基酸分解代谢酶的基因表达。
图2显示丝瓜粉对高脂饮食小鼠血清中支链氨基酸水平的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05)。
图2表明高脂饮食小鼠血清中缬氨酸、亮氨酸、异亮氨酸含量与对照组相比分别显著上升约41%、38%及65%,丝瓜干预组小鼠各指标仅分别上升约22%、9%及20%;总体而言,对照组小鼠血清总支链氨基酸水平是46.32±3.38μg/mL,高脂组小鼠血清中该指标显著上升约44%至66.61±3.93μg/mL,丝瓜干预组小鼠血清中该指标仅上升约16%至53.83±1.98μg/mL;因此,高脂饮食小鼠出现高支链氨基酸血症,体内BCAA水平显著上升,丝瓜粉干预可明显缓解高支链氨基酸血症。
图3、图4显示丝瓜粉对高脂饮食小鼠体重和脂肪分布的影响。其中,小写字母表示在p=0.05水平下进行差异比较,字母不同表示不同处理组间差异显著(p<0.05),字母相同表示不同处理组间差异不显著(p>0.05)。
图3、图4表明与对照组相比,高脂饮食小鼠体重明显增加,全身均有明显的脂肪分布,丝瓜粉干预可明显降低高脂饮食小鼠体重增加,缓解全身脂肪分布。
图5显示丝瓜粉对高脂饮食小鼠脂肪组织脂质堆积的影响。
图5表明与对照组相比,高脂饮食小鼠脂肪组织中细胞体积明显增大,细胞大小不均等,脂质堆积严重,丝瓜粉干预可明显降低高脂饮食小鼠脂肪细胞中脂质积累。
图6显示丝瓜粉对高脂饮食小鼠肝脏组织脂质堆积的影响。
图6表明与对照组相比,高脂饮食小鼠肝脏组织中脂滴大小不一,脂质堆积严重,丝瓜粉干预可明显降低高脂饮食小鼠肝脏细胞中脂质积累。
表1为丝瓜粉对高脂饮食小鼠血生化指标的影响:
对照组 | 高脂组 | 丝瓜干预组 | |
FFA(mmol/L) | 0.35±0.02<sup>b</sup> | 0.46±0.01<sup>a</sup> | 0.37±0.01<sup>b</sup> |
TG(mmol/L) | 0.39±0.01<sup>b</sup> | 0.45±0.24<sup>a</sup> | 0.39±0.05<sup>b</sup> |
TC(mmol/L) | 3.19±0.19<sup>b</sup> | 4.63±0.24<sup>a</sup> | 3.65±0.06<sup>b</sup> |
HDL(mmol/L) | 2.54±0.17<sup>b</sup> | 3.44±0.12<sup>a</sup> | 2.88±0.09<sup>b</sup> |
LDL(mmol/L) | 0.56±0.001<sup>b</sup> | 0.92±0.13<sup>a</sup> | 0.62±0.05<sup>b</sup> |
Glu(mmol/L) | 5.20±0.08<sup>c</sup> | 8.40±0.40<sup>a</sup> | 6.06±0.36<sup>b</sup> |
INS(mIU/L) | 11.45±1.31<sup>b</sup> | 18.38±1.66<sup>a</sup> | 13.02±0.67<sup>b</sup> |
HOMA-IR | 2.65±0.34<sup>b</sup> | 6.92±0.91<sup>a</sup> | 3.54±0.38<sup>b</sup> |
表1表明丝瓜粉显著降低高脂饮食小鼠血清中脂质水平(FFA、TG、TC),减少血清中LDL水平,降低血糖含量和胰岛素含量,改善高脂饮食小鼠的胰岛素抵抗。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,根据本发明的技术方案及其构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (6)
1.一种丝瓜粉在制备用于非治疗目的的改善高支链氨基酸血症营养品中的应用。
2.如权利要求1所述的应用,其特征在于:所述丝瓜粉的制备方法包括如下步骤:
1)丝瓜清洗切片:摘取新鲜丝瓜,清洗干净后切成薄片;
2)低温冷冻干燥:步骤1中的丝瓜薄片在-20℃至-80℃下预冻24h-48h,然后在-60℃至-80℃下低温冷冻干燥48h-72h;
3)超微粉碎:步骤2中经冷冻干燥的丝瓜薄片超微粉碎后过60目筛后得到丝瓜粉。
3.如权利要求2所述的应用,其特征在于:
步骤1中,摘取的新鲜丝瓜清洗干净后切至2-3mm厚度薄片。
4.如权利要求2所述的应用,其特征在于:
步骤2中,步骤1中的丝瓜薄片在-80℃下预冻48h,然后在-80℃下低温冷冻干燥72h。
5.如权利要求2所述的应用,其特征在于:
所述丝瓜粉的制备方法还包括:
4)步骤3制备得到的丝瓜粉,避光-20℃保存。
6.如权利要求2所述的应用,其特征在于:
步骤3中得到的丝瓜粉,使用前用无菌生理盐水混合成0.2 g/mL配制丝瓜悬浊液。
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