CN110897022A - 柑普茶在制备调节肠道菌群作用食品中的应用 - Google Patents
柑普茶在制备调节肠道菌群作用食品中的应用 Download PDFInfo
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Abstract
本发明公开了柑普茶在调节肠道菌群方面的用途。包括柑普茶在制备调节肠道菌群食品方面的用途。及柑普茶在制备调节增加肠道双歧杆菌属或乳酸杆菌属丰度食品中的应用。经实验证明,柑普茶能显著增加健康大鼠和脾虚大鼠肠道菌群中益生菌双歧杆菌属和乳酸杆菌属的相对丰度,降低肠道潜在致病菌的含量,可优化肠道菌群结构,其效果比单独使用普洱茶或广陈皮明显更优。
Description
技术领域
本发明属于食品领域,尤其涉及柑普茶在调节肠道菌群方面的用途。
背景技术
柑普茶是当前养生保健的新兴茶品,是以产于新会的茶枝柑鲜果皮的干品 (或其经陈化后的陈皮),与普洱熟茶经过烘焙、陈化等工艺加工而成。它融合了新会柑清醇的果香味和云南普洱茶醇厚的陈香味,茶性温和甘醇,滋味独特,老少皆宜;且兼具陈皮和普洱茶的保健功效,具有抗炎、抗氧化、降血脂、改善脾胃等多种作用,深受消费者喜爱。
肠道菌群是指生物体肠道中的微生物,主要由细菌、古细菌、真菌、病毒等构成。健康成人的肠道内大概有1014个细菌,其数量是自身细胞总数的10倍,主要有硬壁菌、拟杆菌、放线菌和变形杆菌。研究表明,人体的健康不仅与自身的基因组有关,还与肠道内环境微生物有密不可分的联系。肠道菌群通过与宿主间的相互作用调控肠道稳态,各种原因导致的肠道菌群失调均可导致疾病的发生。因此,保持肠道菌群稳态是维持人体健康所必需的。
目前,未发现柑普茶调节肠道菌群的相关文献报道。
发明内容
本发明的目的是提供柑普茶在调节肠道菌群方面的用途。包括包括柑普茶在制备调节增加肠道双歧杆菌属或乳酸杆菌属丰度食品中的应用。尤其是柑普茶在制备改善脾虚证肠道菌群结构作用食品中的应用。
本发明的柑普茶经实验证明:
1、柑普茶能显著增加健康大鼠肠道菌群中益生菌双歧杆菌属(Bifidobacterium)、乳酸杆菌属(Lactobacillus)的相对丰度,优化肠道菌群结构,其效果比单独使用普洱茶、广陈皮更优;
2、柑普茶能够显著提高脾虚大鼠的肠道菌群物种丰富度,升高双歧杆菌属(Bifidobacterium)等益生菌的丰度,降低Desulfovibrio等潜在致病菌的含量,使肠道菌群结构趋于正常,其效果优于单独使用的普洱茶及广陈皮,即发现柑普茶能够调节肠道菌群这一新用途。
附图说明
图1是本发明实施例柑普茶、普洱茶及广陈皮对健康大鼠肠道菌群结构影响的实验结果图示;
图2是本发明实施例提供各组大鼠肠道菌群的主坐标分析(PCoA)得分图;
图3是本发明实施柑普茶、普洱茶及广陈皮对脾虚大鼠肠道菌群中益生菌相对丰度影响的实验结果图示;
图4是本发明实施例柑普茶、普洱茶及广陈皮对脾虚大鼠肠道菌群中潜在致病菌相对丰度影响的实验结果图示。
具体实施方式
下面结合具体实验对柑普茶在调节肠道菌群方面的新用途做进一步的说明。
1、实验动物
SD大鼠,SPF级,雄性,体重200-250g,购自广东省医学实验动物中心,许可证号:SCXK(粤)2013-0002。
2、柑普茶的提取
将柑普茶粉碎,称取100g,浸泡在沸水中3次(依次为2L、20分钟,1.5 L、15分钟,1.5L、15分钟),趁热过滤,合并滤液,滤液60℃下减压浓缩至500 mL,得到浓度为0.2g/mL的浓缩茶汤。
柑普茶中普洱茶与广陈皮的重量比平均为8:2,取与柑普茶等量同一批次柑普茶的原料普洱茶、广陈皮分别用沸水提取,浓缩为浓度一致的普洱茶、广陈皮提取物。
3、柑普茶对健康大鼠肠道菌群的调节作用
动物分组及给药
取雄性SD大鼠40只,随机分为4组,每组10只,分组及给药剂量如下:
①正常对照组:灌胃给予纯净水(0.5mL/100g,一天两次);
②柑普茶组:灌胃给予柑普茶浓缩茶汤(1.5g/kg,一天两次);
③普洱茶组:灌胃给予普洱茶浓缩茶汤(1.5g/kg,一天两次);
④广陈皮组:灌胃给予广陈皮提取物(1.5g/kg,一天两次)。
连续灌胃28天。
(2)取样及检测
末次灌胃给药后(第28天傍晚),将大鼠禁食12h,自由饮水。次日,腹腔注射10%水合氯醛(0.3ml/100g)麻醉大鼠,纵剖结肠,收集结肠内粪便于1.5mL 冻存管中,立即放入液氮中,随后转移至-80℃保存。采用16S rDNA测序技术,对收集到的粪便样品进行测序,考察柑普茶、普洱茶和广陈皮对健康大鼠肠道菌群的影响。
(3)结果
本实验对四组大鼠肠道菌群测序结果进行了Alpha多样性分析[以ACE、 Chao1、香农指数(Shannon index)和辛普森指数(Simpson index)表示],结果如表1和图1所示,持续28天灌胃给予柑普茶、普洱茶或广陈皮提取物对肠道菌群整体丰度及多样性没有显著影响。进一步的LEfSe分析发现,灌胃给予柑普茶后,健康大鼠肠道菌群中双歧杆菌属(Bifidobacterium)和乳酸杆菌属 (Lactobacillus)相对丰度显著增加,且相对丰度较灌胃给予普洱茶及广陈皮的高。这些结果表明,与普洱茶及广陈皮相比,柑普茶能更好地增加健康个体益生菌的相对丰度,优化肠道菌群的结构。
表1柑普茶、普洱茶及广陈皮对健康大鼠肠道菌群多样性的影响
4、柑普茶对脾虚大鼠肠道菌群的调节作用
(1)动物分组及给药
取雄性SD大鼠50只,随机分为5组(正常对照组、脾虚模型对照组、脾虚大鼠灌胃柑普茶组、脾虚大鼠灌胃普洱茶组、脾虚大鼠灌胃广陈皮组),每组10 只。采用皮下注射利血平(0.5mg/kg/d)造成大鼠脾虚模型(连续14天),正常对照组皮下注射等体积生理盐水。造模后,各组大鼠灌胃给药14天,分组及给药剂量如下:
①正常对照组:灌胃给予纯净水(0.5mL/100g,一天两次);
②脾虚模型对照组:灌胃给予纯净水(0.5mL/100g,一天两次);
③脾虚大鼠灌胃柑普茶组:灌胃给予柑普茶浓缩茶汤(1.5g/kg,一天两次);
④脾虚大鼠灌胃普洱茶组:灌胃给予普洱茶浓缩茶汤(1.5g/kg,一天两次);
⑤脾虚大鼠灌胃广陈皮组:灌胃给予广陈皮提取物(1.5g/kg,一天两次)。
(2)取样及检测
末次灌胃给药后(第14天傍晚),将大鼠禁食12h,自由饮水。次日,腹腔注射10%水合氯醛(0.3ml/100g)麻醉大鼠,纵剖结肠,收集结肠内粪便于1.5mL 冻存管中,立即放入液氮中,随后转移至-80℃保存。采用16S rDNA测序技术,对收集到的粪便样品进行测序,考察柑普茶、普洱茶和广陈皮对脾虚大鼠肠道菌群的影响。
(3)结果
本实验对5组大鼠肠道菌群测序结果进行了Alpha多样性分析,并采用主坐标分析法(Principal coordinates analysis,PCoA)研究各组大鼠肠道菌群的组成差异。结果如表2所示,脾虚模型对照组大鼠肠道菌群的ACE、Chao1、香农指数(Shannon index)均极显著降低,表明其菌群丰富度及多样性都显著降低。图2a为正常对照组与脾虚模型组间的PCoA分析结果,正常对照组与模型组在图上能够完全分开,表明脾虚大鼠肠道菌群已经发生明显的改变。经柑普茶、普洱茶及广陈皮干预后,脾虚大鼠肠道菌群的ACE、Chao1指数显著升高,提示三者均可以显著提高脾虚大鼠肠道菌群的物种丰富度。相比之下,柑普茶组的ACE、Chao1指数较普洱茶组、广陈皮组高,提示这三种饮品中,柑普茶对脾虚大鼠肠道菌群的物种丰富度调节能力更好。图2b为5组数据的分析结果,柑普茶、普洱茶及广陈皮组更多地与正常对照组重叠,相比于模型组,与正常对照组有更近的距离。说明给茶干预对大鼠肠道菌群结构都有一定的影响,干预后菌群结构趋于正常组,与模型组出现差异,提示三者能够通过影响菌群结构对宿主健康产生影响。将肠道菌群相对丰度与脾虚药效指标进行斯皮尔曼(Spearman)相关分析,识别出与脾虚相关的菌属,发现:与正常大鼠相比,脾虚大鼠肠道菌群中双歧杆菌属(Bifidobacterium)、拟普雷沃菌属(Alloprevotella)、阿洛巴氏菌属(Allobaculum)、嗜冷杆菌属(Psychrobacter)、产粪甾醇真细菌([Eubacterium] _coprostanoligenes_group)、uncultured_bacterium_f_Erysipelotrichaceae等益生菌的相对丰度明显降低(如图3所示),而脱硫弧菌属(Desulfovibrio)、颤杆菌克属(Oscillibacter)、Ruminiclostridium_9、另枝菌属(Alistipes)、uncultured_bacterium_o_Gastranaerophilales、副杆状菌属(Parabacteroides)等潜在致病菌的相对丰度则明显升高(如图4所示),提示脾虚大鼠肠道菌群结构失调;经柑普茶干预后,脾虚大鼠肠道菌群中益生菌的相对丰度明显升高,致病菌的相对丰度则明显降低,肠道菌群结构趋于正常,表明柑普茶可以恢复脾虚大鼠的肠道菌群结构,且其效果优于单独使用的普洱茶及广陈皮。
表2柑普茶、普洱茶及广陈皮对脾虚大鼠肠道菌群多样性的影响
注:n=10,与正常对照组相比,**P<0.01;与模型组相比,##P<0.01 。
Claims (3)
1.柑普茶在制备具有调节肠道菌群作用食品中的应用。
2.柑普茶在制备调节增加肠道双歧杆菌属或乳酸杆菌属丰度食品中的应用。
3.柑普茶在制备改善脾虚证肠道菌群结构作用食品中的应用。
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