CN115433703B - 辣椒碱在促进嗜粘蛋白阿克曼氏菌增殖中的应用 - Google Patents
辣椒碱在促进嗜粘蛋白阿克曼氏菌增殖中的应用 Download PDFInfo
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Abstract
本发明提出了辣椒碱在制备药物、食品、保健品或试剂盒中的用途，所述药物、食品、保健品或试剂盒用于促进嗜粘蛋白阿克曼氏菌增殖。辣椒碱可以促进嗜粘蛋白阿克曼氏菌增殖，有助于改善肠道微环境，起到预防或治疗肠道疾病的作用，同时，为辣椒碱新功能的开发和嗜粘蛋白阿克曼氏菌特性研究奠定了理论基础，具有重要意义。
Description
技术领域
本发明涉及生物领域。具体地，本发明涉及辣椒碱在促进嗜粘蛋白阿克曼氏菌增殖中的应用。
背景技术
嗜粘蛋白阿克曼氏菌（Akkermansiamuciniphila）是肠道中一株可以降解肠道粘蛋白的菌株，是由荷兰瓦赫宁根研究人员从健康人群粪便中分离并鉴定获得的，属于Verrucomicrobiota菌门，Akkermansia菌属。大量的研究发现Akkermansiamuciniphila在肥胖、糖尿病、心血管疾病、肠道炎症、肝脏疾病和神经疾病相对丰度降低，而干预实验已证实增加肠道Akkermansiamuciniphila的相对丰度可显著改善肥胖和糖尿病等疾病。
辣椒（Capsicum annum L.）是我国第一大单品蔬菜，2021年种植面积达3375万亩，产量达6000万吨以上，年产值超2500亿元（来源：国家特色产业技术体系）。据FAO统计，中国已经成为世界上最大的辣椒生产国，占世界产量的46 %。辣椒有“蔬菜之冠”的美称，营养价值高，含有丰富的如维生素C、辣椒素、类胡萝卜素、类黄酮和蛋白质。辣椒碱是辣椒呈现辣味的主要成分，已有研究显示辣椒素可参与调控代谢紊乱，具有改善认知功能障碍、减肥、降血压、降血糖和抑制肿瘤等作用。
然而，目前并未有研究报道关于辣椒碱与嗜粘蛋白阿克曼氏菌增殖具有相关性，有待研究。
发明内容
本发明旨在至少在一定程度上解决现有技术中存在的技术问题。为此，本发明提出了辣椒碱在制备药物、食品、保健品或试剂盒中的用途、辣椒碱在体外促进嗜粘蛋白阿克曼氏菌增殖中的应用、培养基、促进嗜粘蛋白阿克曼氏菌增殖的方法，辣椒碱可以促进嗜粘蛋白阿克曼氏菌增殖，有助于改善肠道微环境，起到预防或治疗肠道疾病的作用，同时，为辣椒碱新功能的开发和嗜粘蛋白阿克曼氏菌特性研究奠定了理论基础，具有重要意义。
在本发明的一个方面，本发明提出了辣椒碱在制备药物、食品、保健品或试剂盒中的用途。根据本发明的实施例，所述药物、食品、保健品或试剂盒用于促进嗜粘蛋白阿克曼氏菌增殖。发明人发现，辣椒碱可以促进嗜粘蛋白阿克曼氏菌增殖，有助于改善肠道微环境，起到预防或治疗肠道疾病的作用，同时，为辣椒碱新功能的开发和嗜粘蛋白阿克曼氏菌特性研究奠定了理论基础，具有重要意义。
根据本发明的实施例，所述辣椒碱在制备药物、食品、保健品或试剂盒中的用途还可以具有下列附加技术特征：
根据本发明的实施例，所述辣椒碱来源于鲜辣椒、干辣椒或辣椒制品。
根据本发明的实施例，所述药物、食品或保健品的给予计量选自20~50mg/kg体重/天。由此，可以更好地发挥辣椒碱促进嗜粘蛋白阿克曼氏菌增殖的效果。
根据本发明的实施例，所述药物、食品或保健品的剂型选自片剂、液体、粉末冲剂、咀嚼片剂或软凝胶。
在本发明的另一方面，本发明提出了辣椒碱在体外促进嗜粘蛋白阿克曼氏菌增殖中的应用。辣椒碱可以促进嗜粘蛋白阿克曼氏菌增殖，为辣椒碱新功能的开发和嗜粘蛋白阿克曼氏菌特性研究奠定了理论基础。
在本发明的又一方面，本发明提出了一种培养基。根据本发明的实施例，所述培养基包括：用于培养嗜粘蛋白阿克曼氏菌的基础培养基；以及辣椒碱。根据本发明实施例的培养基中含有辣椒碱，有助于促进嗜粘蛋白阿克曼氏菌增殖。
需要说明的是，本发明对于用于培养嗜粘蛋白阿克曼氏菌的基础培养基组成不做严格限定，可以为本领域常规的用于培养嗜粘蛋白阿克曼氏菌的培养基，例如含有蛋白胨、脱水小牛脑浸粉、脱水牛心浸粉、氯化钠、葡萄糖、磷酸氢二钠等，具体地，可以选自CM1135B脑心浸出液肉汤培养基。
根据本发明的实施例，所述辣椒碱来源于鲜辣椒、干辣椒或辣椒制品。
在本发明的又一方面，本发明提出了一种促进嗜粘蛋白阿克曼氏菌增殖的方法。根据本发明的实施例，所述方法包括：使嗜粘蛋白阿克曼氏菌在含有辣椒碱的培养基中培养。由此，根据本发明实施例的方法有助于促进嗜粘蛋白阿克曼氏菌增殖，提高嗜粘蛋白阿克曼氏菌产量。
需要说明的是，本发明对于含有辣椒碱的培养基中除辣椒碱以外的其他组成不做严格限定，可以为本领域常规的用于培养嗜粘蛋白阿克曼氏菌的基础培养基组成，对于培养基中辣椒碱的含量、嗜粘蛋白阿克曼氏菌的接种量、培养条件也不做严格限定，可以根据实际需要灵活选择。例如，基础培养基中辣椒碱含量可以为20ng/mL~1mg/mL，嗜粘蛋白阿克曼氏菌是以菌液的形式提供，菌液浓度为106~108CFU/mL，接种量为5%~25%，培养温度为30℃~40℃，时间为20h~60h。
根据本发明的实施例，所述辣椒碱来源于鲜辣椒、干辣椒或辣椒制品。
本发明的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本发明的实践了解到。
附图说明
本发明的上述和/或附加的方面和优点从结合下面附图对实施例的描述中将变得明显和容易理解，其中：
图1为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群α多样性对比分析图（每组7只）；
图2为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群主成分分析图（每组7只）；
图3为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群门水平相对丰度对比分析图（每组7只）；
图4为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群属水平相对丰度对比分析图（每组7只）；
图5为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群种水平相对丰度对比分析图（每组7只）。
具体实施方式
下面将结合实施例对本发明的方案进行解释。本领域技术人员将会理解，下面的实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例1
1、实验材料
本发明所用的辣椒碱购自上海同田生物技术有限公司，分子式：C18H27NO3，分子量：305.4，CAS号：404-86-4，检测纯度（HPLC）：98%。
CM1135B脑心浸出液肉汤培养基。
嗜粘蛋白阿克曼氏菌（Akkermansia）：购自德国微生物菌种保藏中心DSMZ，编号为DSM 22959
2、步骤
实验组：按照DSMZ提供的实验步骤说明书活化A. muciniphila冻干粉，添加0.5mL脑心浸出液肉汤培养基至安培瓶中，在H2/CO2/N2（5/5/90，v/v/v）气相条件下，37℃培养30min。将活化的菌液在脑心浸出液肉汤固体培养基中划线培养24h-48h，沾取单菌落在脑心浸出液肉汤培养基中扩大培养。
将上步扩大培养所得菌液以10%的接种量接种于脑心浸出液肉汤培养基中，于37℃、H2/CO2/N2（5/5/90，v/v/v)条件下培养24h，连续转接2次后，取培养至对数期的菌接种至预先制备的含不同浓度辣椒碱（20ng/mL-1mg/mL）的脑心浸液琼脂培养基，36h-60h后菌落计数。
对照组：同实验组，区别在于培养基中不添加辣椒碱。
对培养产物进行菌落计数，结果显示，实验组的菌落数显著高于对照组，由此表明辣椒碱的添加可以显著促进嗜粘蛋白阿克曼氏菌增殖。
实施例2
选取4月龄的APP/PS1雄鼠，并在标准12小时昼夜循环、22℃恒温、无特定病原体(SPF)环境下进行饲养。在适应性饲养2周后，随机将小鼠分成2组，分别为对照组（APP/PS1_CTRL）和处理组(APP/PS1_CP) ，0.5%羧甲基纤维素钠作为溶剂溶解辣椒碱，处理组小鼠每天灌胃40mg/kg的辣椒碱，对照组给予0.5%羧甲基纤维素钠作为空白对照，持续给药4个月后，搜集小鼠粪便，利用16S rRNA测序技术分析辣椒碱对APP/PS1小鼠肠道微生物菌群结构的影响。
2组不同处理的APP/PS1小鼠的α多样性结果如图1所示，与APP/PS1_CTRL组小鼠相比，APP/PS1_CP组Shannon指数显著降低（P = 0.0413），Simpson指数显著增加（P =0.0295），这说明经辣椒碱处理的APP/PS1小鼠的肠道菌群多样性显著降低，这可能与辣椒碱具有杀菌的作用有关。
对2个不同处理组的β多样性分析，主成分分析的结果如图2所示。结果显示APP/PS1_CTRL组和APP/PS1_CP组出现明显聚类且显著分开，表明辣椒碱干预后APP/PS1小鼠的肠道菌群发生显著变化。
图3为APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群在门水平上的对比图。由图3可知，经辣椒碱处理后，APP/PS1小鼠的Verrucomicrobiota增加，而Bacteroidota、Firmicutes和Actinobacteriota均有所降低，但没有显著差异。
APP/PS1_CTRL组和APP/PS1_CP组小鼠肠道菌群在属水平上对比分析如图4所示。结果表明APP/PS1_CP组小鼠肠道中Akkermansia显著增加，即辣椒碱促进APP/PS1小鼠肠道中Akkermansia的增殖。
图5为2组在种水平上相对丰度的对比分析图。与APP/PS1_CTRL组相比，APP/PS1_CP组Akkermansiamuciniphila显著增加（P = 0.0002）。其中APP/PS1_CTRL组小鼠肠道中Akkermansiamuciniphila的比例仅为0.061%，辣椒碱灌胃处理后，Akkermansiamuciniphila在APP/PS1小鼠肠道中增至12.03%。即辣椒碱显著促进了APP/PS1小鼠肠道中Akkermansiamuciniphila菌株的增殖。
尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制，本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
Claims (3)
1.辣椒碱在制备药物、食品、保健品或试剂盒中的用途，其特征在于，所述药物、食品、保健品或试剂盒用于促进APP/PS1鼠体内嗜粘蛋白阿克曼氏菌增殖。
2.根据权利要求1所述的用途，其特征在于，所述辣椒碱来源于鲜辣椒、干辣椒或辣椒制品。
3.根据权利要求1所述的用途，其特征在于，所述药物、食品或保健品的给予计量选自20~50mg/kg体重/天；
所述药物、食品或保健品的剂型选自片剂、液体、粉末冲剂、咀嚼片剂或软凝胶。
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