CN113974159A - 齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用 - Google Patents
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Abstract
本发明涉及微生物技术领域,具体公开了齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用。本申请通过体外模拟肠道环境,并以健康人体粪便中收集的肠道菌群作为样本,将齿叶乳香树树脂应用到肠道有益菌增殖发酵中,发现齿叶乳香树树脂可以显著促进肠道有益菌的增殖,尤其针对乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌等肠道有益菌的增殖促进效果极其显著,此外,本申请还提供了齿叶乳香树树脂在制备用于治疗或改善肠道炎症性疾病的药物中的应用。本申请为临床应用提供科学依据,扩大了齿叶乳香树树脂的应用领域。
Description
技术领域
本发明涉及微生物技术领域,具体而言,涉及齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用。
背景技术
齿叶乳香树树脂是一种从齿叶乳香树上提取的树胶树脂,主产于红海沿岸,具有抗炎、抗肿瘤、抗溃疡、抗哮喘、抗氧化、镇痛、免疫调节和调控脂质代谢等药理作用。齿叶乳香树树脂在化妆品、护肤品里有着多种功效。对过氧化酶激活受体有很好的活化作用,在皮肤损伤愈合过程中起着重要作用,具有抗炎作用。可增强脂肪酶活性,助于脂肪分解,具有控油抗脂溢作用。可以延长细胞生命,具有抗衰作用。此外,齿叶乳香树树脂可以有效地抑制胰腺、结肠、直肠中的肿瘤生长,是一种非常有前景的抗癌草药。然而其对于肠道微生物的调节作用,目前鲜有相关报道。
人体肠道中存在的数量庞大的微生物,它们发挥的功能多种多样,包括物质代谢、生物屏障、免疫调控及宿主防御等。肠道微生物不仅帮助人体从食物中吸收营养,还能够合成氨基酸、有机酸、维生素、抗生素等供我们利用,并可以将产生的毒素加以代谢,减少对人体的毒害。肠道微生物和人体存在着互利共生的关系,对于维持人的健康发挥着重要的作用。
鉴于此,特提出本发明。
发明内容
本发明的目的在于提供齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用,经研究发现,齿叶乳香树树脂可以有效促进肠道有益菌的增殖,从而调节肠道菌群,起到改善或治疗肠道炎症性疾病的作用。
本发明是这样实现的:
第一方面,本发明提供齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用。
在可选的实施方式中,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、布劳特氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌和长栖粪杆菌;
优选地,所述肠道有益菌为乳杆菌。
在可选的实施方式中,所述齿叶乳香树树脂的用量为1.49-1.52g/15ml。
第二方面,本发明提供一种促进肠道有益菌增殖的产品,其包括作为唯一活性成分的齿叶乳香树树脂。
在可选的实施方式中,所述促进肠道有益菌增殖的产品包括改善肠道菌群的食品或保健品。
第三方面,本发明提供齿叶乳香树树脂在制备用于治疗或改善肠道炎症性疾病的药物中的应用。
在可选的实施方式中,所述肠道炎症性疾病包括肝硬化和便秘中的至少一种。
第四方面,本发明提供一种促进肠道有益菌增殖的方法,其包括在培养肠道有益菌的培养基中加入齿叶乳香树树脂。
在可选的实施方式中,所述齿叶乳香树树脂的加入量为1.49-1.52g/15ml。
本发明具有以下有益效果:
本申请通过体外模拟肠道环境,并以健康人体粪便中收集的肠道菌群作为样本,将齿叶乳香树树脂应用到肠道有益菌增殖发酵中,发现齿叶乳香树树脂可以显著促进肠道有益菌的增殖,尤其针对乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌等肠道有益菌具有较佳的促进效果,本申请为临床应用提供科学依据,达到高效制备促进肠道有益菌增殖的产品,同时还可以通过向常规培养基中加入齿叶乳香树树脂以促进肠道有益菌增殖。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,应当理解,以下附图仅示出了本发明的某些实施例,因此不应被看作是对范围的限定,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他相关的附图。
图1为本申请实施例提供的肠道微生物样品经齿叶乳香树树脂干预前后的丰度变化示意图。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
本申请提供了齿叶乳香树树脂的新用途。齿叶乳香树树脂是从齿叶乳香树中提取获得的,其在化妆品、护肤品里主要作用是抗氧化剂,控油抗脂溢,抗炎剂。对过氧化酶激活受体有很好的活化作用,在皮肤损伤愈合过程中起着重要作用,说明有很好的抗炎性;可增强脂肪酶活性,助于脂肪分解,可在减肥产品或油性皮肤的洗涤用品中使用;对半胱天冬蛋白酶的活化有促进作用,可以延长细胞生命,具有抗衰作用;同时有可以促进毛发的生长,是不错的毛发生长剂。
本申请创新性的提出了齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用。
肠道有益菌有多种,经研究发现,齿叶乳香树树脂对乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、布劳特氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种具有促进其增殖的作用。
其中,上述肠道有益菌对应的英文名分别为:乳杆菌(Lactobacillus)、长栖粪杆菌(Faecalibacterium)、粪厌氧棒杆菌(Anaerostipes)、副普氏菌(Paraprevotella)、布劳特氏菌(Blautia)、梭菌XVIII(Clostridium XVIII)和梭状芽孢杆菌(Fusicatenibacter)。
齿叶乳香树树脂尤其对乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种的增殖促进效果显著。而对布劳特氏菌具有一定的促进增殖作用,但是效果无明显变化。
进一步地,根据促进增殖的效果,本申请中,肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌中的至少一种;
优选地,肠道有益菌包括乳杆菌和长栖粪杆菌,齿叶乳香树树脂可以同时对乳杆菌和长栖粪杆菌具有较佳的促进增殖效果;更优选地,肠道有益菌为乳杆菌;齿叶乳香树树脂对乳杆菌的增殖促进率大于200%。
本申请的齿叶乳香树树脂购于西安奥赛生物科技有限公司,纯度60%。经发明人研究发现,当齿叶乳香树树脂的用量达到1.49-1.52g/15ml时其具有较佳的促进肠道有益菌增殖的作用。
众所周知,肠道有益菌的失调会导致众多疾病,例如,在文献“Strati,F.,Cavalieri,D.,Albanese,D.,De Felice,C.,Donati,C.,Hayek,J.,…De Filippo,C.(2016).Altered gut microbiota in Rett syndrome.Microbiome,4(1)”中公开了上述肠道益生菌的失调可能导致便秘。文献“Fujimoto,T.,Imaeda,H.,Takahashi,K.,Kasumi,E.,Bamba,S.,Fujiyama,Y.,&Andoh,A.(2013).Decreased abundance of Faecalibacteriumprausnitziiin the gut microbiota of Crohn’s disease.Journal ofGastroenterology and Hepatology,28(4),613–619.”公开了肠道益生菌的失调可能导致克罗恩疾病。
由于本申请研究发现,齿叶乳香树树脂具有促进肠道有益菌(例如:乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、布劳特氏菌、梭菌XVIII和梭状芽孢杆菌等)增殖的效果,因此,本申请还可以将齿叶乳香树树脂应用于制备肠道炎症性疾病的药物中,具体来说,肠道炎症性疾病包括结肠癌、克罗恩和便秘中的至少一种。
进一步地,本发明提供一种促进肠道有益菌增殖的产品,其包括作为唯一活性成分的齿叶乳香树树脂。本申请将齿叶乳香树树脂作为唯一活性成分,其可以有效促进肠道有益菌的增殖,有利于调节肠道菌群,为临床应用提供科学依据,达到高效制备促进肠道有益菌增殖的产品。上述促进肠道有益菌增殖的产品还可以添加在食品或保健品中以改善肠道菌群。
此外,本发明还提供了一种促进肠道有益菌增殖的方法,其包括在培养肠道有益菌的培养基中加入齿叶乳香树树脂,南非醉茄提取物的加入量为1.49-1.52g/15ml。通过在特定的肠道有益菌培养发酵时,加入齿叶乳香树树脂,可以有效提升肠道有益菌的活菌数。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
实施例
在体外进行体内模拟实验,取人体粪便作为肠道菌群样品,观察齿叶乳香树树脂干预对于肠道菌群的作用,具体操作过程如下:
1、粪便样品处理
(1)PBS缓冲液配制
以1000ml为例,用电子分析天平分别称取下列药品,倒入1000ml烧杯中,包括:KH2PO4 0.24g,Na2HPO4·12H2O 2.90g,NaCl 8.00g,KCl 0.20g。
用量筒量取800ml超纯水倒入烧杯中,放入搅拌转子,并放到磁力加热搅拌器上搅拌至完全溶解,然后用校准后的PH计测量上述溶液的PH,用0.1M的HCl或NaOH调整PH=7.4±0.05。
将上述完全溶解并调节pH后的溶液用玻璃棒引流至1000ml容量瓶中,用少量超纯水冲洗烧杯后倒入容量瓶,重复3次。将容量瓶中的液体用超纯水定容至刻度线,塞上盖子后上下颠倒10次,确保溶液充分混匀。
将定容后的溶液倒入洁净黄色盖子的玻璃瓶,每1000ml的瓶子内放500ml溶液,防止高压灭菌时喷出。将黄色玻璃瓶盖子拧松后,放入高压灭菌锅,121℃,15分钟。取出后迅速拧紧盖子。待降至室温后,将溶液需存于4℃冰箱中,可保存6个月。
(2)粪便样品处理
以10g为例,最终浓度为10%。
用百分之一的天平称量出10g粪便样品,用自动移液枪取适量上述PBS缓冲液加至离心管中,在振荡器上充分混匀。然后将完全混匀的样品平均分装至新50ml离心管中,每管再加适量PBS缓冲液。
混匀后在生物安全柜内进行过滤,把稀释好的样品依次通过20目、50目、100目、200目的滤网。收集滤液至离心管中,将离心管放入离心机,配平,6000G,4℃离心15min,弃上清。将沉淀称量之后,以终浓度为5%用PBS溶液定容,得到肠道微生物样品。
2、齿叶乳香树树脂干预实验
(1)基础培养基配制
配制用于培养肠道微生物的基础培养基:GAM培养基,以1000ml为例。用电子分析天平称取60g改良GAM肉汤药品,倒入1000ml烧杯中。用量筒量取800ml超纯水倒入烧杯中,放入搅拌转子,并放到磁力加热搅拌器上搅拌至完全溶解。将上述完全溶解的溶液用玻璃棒引流至1000ml容量瓶,并用少量超纯水冲洗烧杯后倒入容量瓶,重复3次。将容量瓶中的液体用超纯水定容至刻度线,塞上盖子后上下颠倒10次,确保溶液充分混匀。
将定容后的溶液倒入洁净黄色盖子的玻璃瓶,每1000ml的瓶子内放500ml培养基,防止高压灭菌时喷出。将装有培养基的黄色盖子的玻璃瓶,瓶盖拧松,放入脉冲高压灭菌锅,使用液体程序,121℃,15分钟。灭菌完后,立即把瓶盖拧紧,冷却至室温。
(2)齿叶乳香树树脂-GAM培养基配制
于无菌操作台上,用1ml的移液枪取上述GAM培养基分装至玻璃管中,每管2ml。将黑色盖子放在瓶口上微掩。接着用1ml移液器取齿叶乳香树树脂溶液(齿叶乳香树树脂购于西安奥赛生物科技有限公司,纯度60%,用PBS缓冲液溶解至质量百分浓度为:5%),加入上述装有GAM培养基的玻璃管中,每管2ml,作为试验组。同时还设置有空白对照组,每组3个重复,分组具体如下:
试验组:2mlGAM培养基+2ml齿叶乳香树树脂溶液+1mLPBS缓冲液;
对照组:2mlGAM培养基+3mLPBS缓冲液;
确认好每个玻璃管盖拧紧后,转移至厌氧箱传递箱中,放置之前需用84消毒液消毒处理。放入厌氧箱后,拧松瓶盖,置换氧气,需置换12个小时。
将上述处理后得到的肠道微生物样品分别接种至试验组(齿叶乳香树树脂-GAM培养基-PBS缓冲液)以及空白对照组(GAM培养基-PBS缓冲液)中,每管1ml。
于厌氧箱中培养72小时后,观察菌群的生长状况,拍照留存。
接着分别将试验组样品和对照组样品,10000rpm,3min离心,弃上清,将沉淀用液氮处理,并送至武汉艾康健生物科技有限公司,利用16S rDNA二代测序法分别测定试验组和对照组中的肠道菌群丰度,即不同肠道微生物在检测到的所有菌种中所占的百分比,其中部分肠道菌群的丰度测定结果如表1所示。
表1不同处理组的肠道菌群丰度
本申请对部分肠道菌群经齿叶乳香树树脂干预前后的丰度变化进行显著性分析。分析结果如图1所示,其中**表示P<0.01,即丰度变化有极显著差异;*表示P<0.05,即丰度变化有显著差异。Lactobacillus、Faecalibacterium、Anaerostipes、Paraprevotella升高极其显著,Clostridium XVIII、Fusicatenibacter有明显增长的趋势,其丰度水平有明显的升高,达到促进肠道有益菌增值的作用,而Blautia无明显变化。
综上所述,本申请通过体外模拟肠道环境,并以健康人体粪便中收集的肠道菌群作为样本,将齿叶乳香树树脂应用到肠道有益菌增殖发酵中,发现齿叶乳香树树脂可以显著促进肠道有益菌的增殖,尤其针对乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌等肠道有益菌的增殖促进效果极其显著,本申请为临床应用提供科学依据,达到高效制备肠道有益菌剂。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (9)
1.齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用。
2.根据权利要求1所述的齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用,其特征在于,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、布劳特氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌、副普氏菌、梭菌XVIII和梭状芽孢杆菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌、长栖粪杆菌、粪厌氧棒杆菌和副普氏菌中的至少一种;
优选地,所述肠道有益菌包括乳杆菌和长栖粪杆菌;
优选地,所述肠道有益菌为乳杆菌。
3.根据权利要求1所述的齿叶乳香树树脂在制备促进肠道有益菌增殖的产品中的应用,其特征在于,所述促进肠道有益菌增殖的产品中所述齿叶乳香树树脂的用量为1.49-1.52g/15ml。
4.一种促进肠道有益菌增殖的产品,其特征在于,其包括作为唯一活性成分的齿叶乳香树树脂。
5.根据权利要求4所述的促进肠道有益菌增殖的产品,其特征在于,所述促进肠道有益菌增殖的产品包括改善肠道菌群的食品或保健品。
6.齿叶乳香树树脂在制备用于治疗或改善肠道炎症性疾病的药物中的应用。
7.根据权利要求6所述的齿叶乳香树树脂在制备用于治疗或改善肠道炎症性疾病的药物中的应用,其特征在于,所述肠道炎症性疾病包括肝硬化和便秘中的至少一种。
8.一种促进肠道有益菌增殖的方法,其特征在于,其包括在培养肠道有益菌的培养基中加入齿叶乳香树树脂。
9.根据权利要求8所述的促进肠道有益菌增殖的方法,其特征在于,所述齿叶乳香树树脂的加入量为1.49-1.52g/15ml。
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