CN112190600A - 一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用 - Google Patents
一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用 Download PDFInfo
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- CN112190600A CN112190600A CN202010954814.4A CN202010954814A CN112190600A CN 112190600 A CN112190600 A CN 112190600A CN 202010954814 A CN202010954814 A CN 202010954814A CN 112190600 A CN112190600 A CN 112190600A
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- lactobacillus plantarum
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- inflammation
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Abstract
本发明涉及微生物领域,公开了一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用。该植物乳杆菌命名为1701,已在2019年10月23日保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号为CGMCC No.18728,微生物分类命名为植物乳杆菌Lactobacillus plantarum。本发明所采用的植物乳杆菌能同时调节细胞因子IL‑12、抑炎因子IL‑10和促炎因子(包括IL‑1β、IL‑6、TNF‑α、MCP‑1和NO)的水平,因而能适应不同原因引起的慢性炎症,并能更全面地防止由慢性炎症引发的疾病。
Description
技术领域
本发明涉及微生物领域,尤其涉及一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用。
背景技术
近年来,随着人们生活水平的不断提高,也涌现出一系列健康问题,亚健康问题日益凸显。导致亚健康的主要原因有:饮食不合理、作息不规律、缺乏运动、睡眠不足、心理压力大、精神紧张、长期不良情绪等。研究发现,不同亚健康患者,体内多多少少会伴随着慢性炎症的发生,并伴随着MCP-1的产生,从而促进巨噬细胞浸润和随后的炎症因子分泌(如IL-10、IL-6、TNF-α和NO等)。一旦巨噬细胞被激活,激活的巨噬细胞会释放炎症细胞因子,从而触发其他免疫细胞(如中性粒细胞,CD8+和CD4+T细胞等)的浸润。在缺乏分解力的情况下,这些细胞因子可能会进入全身循环,促进新陈代谢紊乱,引发长期慢性炎症,影响健康。此外,高血压、动脉粥样硬化、2型糖尿病等疾病也会引发慢性炎症。
一些临床研究已证明,益生菌在预防、改善炎症和代谢紊乱方面具有有益作用,其能够增加乳酸菌和双歧杆菌等有益菌群丰度,降低革兰氏阴性菌丰度,改善肠道微生态环境。 Bifidobacterium animalis ssp.lactis 420能够显著降低血清脂多糖水平和肝脏炎症,以及肠道内大肠杆菌的黏附,从而改善炎症状态。Lactobacillus reuteri GMNL-263活菌和灭活菌株能够显著恢复肠道微生物中有益细菌的降低,提高肠道屏障,下调脂肪和肝脏组织促炎相关基因表达,有效抑制脂肪组织巨噬细胞浸润,改善代谢相关慢性炎症。Lactobacillus rhamnosus Lb102和Bifidobacterium animalis ssp.lactis Bf141可减轻不合理饮食诱导的相关炎症,同时增强肠道完整性标志物,改善肠道微生物菌群组成。但这些菌株具有剂量和菌株特异性,并且个体肠道菌群及生理状态差异明显,不同菌株作用效果存在差异,因此,仍然需要开发有具有缓解慢性炎症的其它益生菌新菌株。此外,现有的植物乳杆菌所能调节的炎症因子种类有限,但由于在不同原因引发的慢性炎症中,发生变化的炎症因子种类不同,而不同的炎症因子对机体的影响存在差异,故现有的植物乳杆菌所能适应的慢性炎症有限。
申请人在前期的研究中从西藏采集的酸奶粉样样品中分离得到一株植物乳杆菌(Lactobacillus plantarum)新菌株1701,并就其在减肥中的应用进行了研究,但是对于该菌株是否具有缓解慢性炎症的能力以及能否具有更多的生理活性,则有待进一步研究。
发明内容
为了解决上述技术问题,本发明提供了一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用。本发明所采用的植物乳杆菌能同时调节细胞因子IL-12、抑炎因子IL-10和促炎因子(包括IL-1β、IL-6、TNF-α、MCP-1和NO)的水平,因而能适应不同原因引起的慢性炎症,并能更全面地防止由慢性炎症引发的疾病。
本发明的具体技术方案为:
一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用,所述组合物含有植物乳杆菌 1701和/或其突变体;所述植物乳杆菌1701已在2019年10月23日保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号为CGMCC No.18728,保藏单位地址为北京市朝阳区北辰西路1号院3号,微生物分类命名为植物乳杆菌Lactobacillus plantarum;所述突变体为对所述植物乳杆菌进行诱变、驯化、基因重组或者经自然突变而获得的突变体。
上述植物乳杆菌1701分离自我国西藏自治区日喀则市乡村采集的酸奶粉样,其16S rRNA基因序列如SEQ ID NO:1所示。
经体外细胞实验证明,上述植物乳杆菌1701及其组合物具有以下功能:
(1)能有效促进脾淋巴细胞增殖,使其提高80.18~125.00%;
(2)能有效促进脾淋巴细胞分泌免疫调节因子IL-12和抑炎因子(即具有抑制炎症作用的细胞因子)IL-10,分别是对照商业菌株鼠李糖乳杆菌GG(LGG)的1.59倍和1.54倍;
(3)能有效促进巨噬细胞分泌抑炎因子IL-10,并抑制巨噬细胞分泌促炎因子(即具有促进炎症作用的细胞因子)IL-6、TNF-α和NO。
经大鼠体内实验证明,上述植物乳杆菌1701的活菌株和灭活菌株在服用浓度 1×109CFU/d时均具有以下功能:
(1)能有效提高血清中抑炎因子IL-10的水平,并降低血清中促炎因子IL-1β、IL-6、TNF-α、 MCP-1和NO的水平;
(2)能有效降低肝脏组织中促炎因子IL-6、IL-1β和NO的水平。
综上所述,植物乳杆菌1701的活菌株和灭活菌株均能同时调节体内多种炎症因子的水平,包括抑炎因子IL-10,以及促炎因子IL-1β、IL-6、TNF-α、MCP-1和NO,并能促进脾淋巴细胞分泌免疫调节因子IL-12,相较于现有的植物乳杆菌而言,其作用更加全面(现有的植物乳杆菌只对其中一种或几种炎症因子有调节作用,无法同时调节上述所有炎症因子),由于在不同原因引起的慢性炎症中,发生变化的炎症因子种类不同,不同炎症因子对机体的影响也存在差异,故植物乳杆菌1701能缓解更多类型的慢性炎症,并能更全面地防止由慢性炎症引发的疾病。
作为优选,所述植物乳杆菌1701为活菌或死菌。
本发明采用的植物乳杆菌1701的灭活菌株也具有缓解慢性炎症的功能,因而组合物中的菌株可为活菌或死菌,组合物的稳定性更好,保质期更长。
作为优选,所述组合物还含有生理上可接受的赋形剂和/或稀释剂。
作为优选,所述组合物为食品、药品、保健品或发酵剂。
作为优选,所述食品为发酵乳、乳酪、含乳饮料、乳粉、固体饮料或发酵果蔬。
作为优选,所述药品的制剂形式为胶囊、粉剂或片剂。
作为优选,所述保健品的制剂形式为胶囊、粉剂或片剂。
作为优选,所述组合物为口服形式。
与现有技术相比,本发明具有以下优点:
(1)本发明所使用的植物乳杆菌1701能促进脾淋巴细胞增殖和分泌免疫调节因子IL-12和抑炎因子IL-10;
(2)本发明所使用的植物乳杆菌1701能促进巨噬细胞分泌抑炎因子IL-10,抑制巨噬细胞分泌促炎因子IL-6、TNF-α和NO;
(3)本发明所使用的植物乳杆菌1701能提高血清中抑炎因子IL-10的水平,降低促炎因子 IL-1β、IL-6、TNF-α、MCP-1和NO的水平;
(4)本发明所使用的植物乳杆菌1701能有效降低肝脏组织中促炎因子IL-6、IL-1β和NO的水平;
(5)本发明所使用的植物乳杆菌1701对免疫调节因子IL-12和抑炎因子IL-10以及促炎因子 IL-1β、IL-6、TNF-α、MCP-1和NO均具有调节作用,因而能适应不同原因引起的慢性炎症,并能更全面地防止由慢性炎症引发的疾病;
(6)本发明所使用的植物乳杆菌1701的灭活菌株也具有缓解慢性炎症的功能,因此采用该菌株制得的组合物具有更好的稳定性和更长的保质期。
附图说明
图1为植物乳杆菌1701发酵乳产酸曲线。
具体实施方式
下面结合实施例对本发明作进一步的描述。
实施例1
本发明所提供的菌株经鉴定属于植物乳杆菌(Lactobacillus plantarum),命名为1701菌株,于2019年10月23日在中国微生物菌种保藏管理委员会普通微生物菌种保藏中心保藏,微生物保藏编号为CGMCC NO.18728。
本发明所提供的菌株是发明人从我国西藏自治区日喀则市乡村采集的酸奶粉样中分离得到。
本发明菌株植物乳杆菌1701的16S rRNA基因序列如SEQ ID NO:1所示。
实施例2
昆明小鼠饲养温度为21±2℃,湿度为30-70%,12h光照交替,自由摄入小鼠维持饲料和饮水。昆明小鼠饲养1周后,脱颈椎处死,无菌取小鼠脾脏,用无菌的玻璃注射器芯将其碾碎, 200目金属筛网过滤后,经ACK细胞裂解液裂解5min,加入含10%胎牛血清的无菌Hank’s 液终止裂解,1000rpm,4℃离心5min,沉淀重悬于5mL含10%胎牛血清的RPMI-1640培养基。用台盼蓝染色,血细胞计数板计数,计算活细胞数和活细胞率。调整细胞浓度为5×106cells/mL。
本发明菌株植物乳杆菌1701和对照商业菌株鼠李糖乳杆菌GG(LGG)经二代活化后,调整菌浓度到1×107CFU/mL。
将细胞悬液加入96孔细胞培养板,每个处理5个重复,分为调零组(细胞培养基),空白对照组(细胞培养基+细胞悬液),诱导剂组(细胞培养基+细胞悬液+诱导剂10μg/mLCon A或2.5μg/mL LPS),菌处理组(细胞培养基+细胞悬液+诱导剂10μg/mL Con A或2.5μg/mL LPS+菌悬液1×107CFU/mL),37℃5%CO2培养箱中培养72h。培养结束后,加入MTT溶液(2.5mg/mL),37℃显色4h后,吸出上清,再加入DMSO。用酶标仪于490nm下测定吸光值。
结果如表1所示,植物乳杆菌1701能够显著促进脾淋巴细胞增殖。在没有诱导剂的条件下,提高125.00%,与对照商业菌株相当,是对照商业菌株的1.05倍;在Con A诱导的条件下,提高114.07%,是对照商业菌株的1.30倍;在LPS诱导的条件下,提高80.18%,与对照商业菌株相当,是对照商业菌株的1.05倍。说明植物乳杆菌1701具有免疫调节活性,且优于对照商业菌株。
表1植物乳杆菌1701促脾淋巴细胞增殖结果
诱导剂 | CK | LGG | 植物乳杆菌1701 |
不加诱导剂 | 0.072±0.001 | 0.154±0.008** | 0.162±0.007** |
ConA(10μg/mL) | 0.135±0.014 | 0.223±0.023** | 0.289±0.029** |
LPS(2.5μg/mL) | 0.111±0.007 | 0.190±0.012** | 0.200±0.012** |
与对照相比,*:p<0.05;**:p<0.01。
实施例3
按照实施例2中小鼠脾淋巴细胞的制备方法制备脾淋巴细胞悬液,调整细胞浓度为5×106 cells/mL。
本发明菌株植物乳杆菌1701和对照商业菌株鼠李糖乳杆菌GG(LGG)经二代活化后,调整菌浓度到1×107 CFU/mL。
将细胞悬液加入96孔细胞培养板,每个处理5个重复,分为调零组(细胞培养基),空白对照组(细胞培养基+细胞悬液),菌处理组(细胞培养基+细胞悬液+菌悬液1×107CFU/mL),37℃5%CO2培养箱中培养48 h。培养结束后,1500 rpm离心10 min,吸取上清,0.22μm滤膜过滤,采用ELISA试剂盒测定IL-10和IL-12含量。
结果如表2所示,植物乳杆菌1701能够显著促进脾淋巴细胞分泌细胞因子IL-10,到达662.28 pg/mL,显著高于商业对照菌株(431.19 pg/mL),是对照商业菌株的1.54倍;同时,植物乳杆菌1701能够显著促进脾淋巴细胞分泌细胞因子IL-12,到达32.64 pg/mL,显著高于商业对照菌株(18.28 pg/mL),是对照商业菌株的1.59倍。说明植物乳杆菌1701能够显著促进脾淋巴细胞分泌细胞因子,且分泌IL-10的能力更强,具有抗炎活性。
表2植物乳杆菌1701促脾淋巴细胞分泌细胞因子结果
处理 | CK | LGG | 植物乳杆菌1701 |
IL-10浓度(pg/mL) | 37.86±2.51<sup>a</sup> | 431.19±30.50<sup>b</sup> | 662.28±43.06<sup>c</sup> |
IL-12浓度(pg/mL) | 4.88±0.35<sup>a</sup> | 18.28±1.67<sup>b</sup> | 32.64±2.08<sup>c</sup> |
IL-10/IL-12 | 7.76 | 23.59 | 20.29 |
a,b,c:p<0.05。
实施例4
建立RAW264.7细胞培养体系,细胞生长于含10%胎牛血清的DMEM培养基中(含青霉素100U/mL,链霉素100mg/mL)。待细胞传至第三代,采用0.25%的胰酶(含EDTA)消化,获得单细胞悬液,细胞以1×106细胞/孔的密度接种于24孔细胞培养板中,于37℃,5%CO2培养箱中培养2 d。
本发明菌株植物乳杆菌1701和对照商业菌株(鼠李糖乳杆菌GG,LGG)经二代活化后,取对数生长末期菌液,4000 rpm离心10 min,弃上清,获得菌泥,重悬于含10%胎牛血清的DMEM完全培养基(不加双抗)中,制成1×109 CFU/mL的菌液。非炎症模型组每孔加入100μL菌悬液,炎症模型组每孔加入100μL菌悬液和10μg/mL LPS,于37℃,5%CO2培养箱中共孵育24 h。然后,1500 rpm离心10 min,吸取上清,0.22μm滤膜过滤,采用ELISA 试剂盒测定IL-10、IL-6、TNF-α和NO含量。
结果如表3所示,在没有炎症诱导剂LPS的条件下,与商业菌株相比,植物乳杆菌1701 促进RAW264.7细胞分泌IL-10的量显著高于商业菌株(p<0.05),分泌NO的量显著低于商业菌株(p<0.05),IL-6和TNF-α分泌量与商业菌株相当。在LPS诱导的炎症条件下,与商业菌株相比,植物乳杆菌1701促进RAW264.7细胞分泌IL-10的量显著高于商业菌株 (p<0.05),分泌IL-6的量显著低于商业菌株(p<0.05),NO和TNF-α分泌量与商业菌株相当。表明在炎症条件下,本发明菌株植物乳杆菌1701能够促进IL-10分泌,抑制IL-6、TNF-α和NO分泌,具有改善炎症的功能。
表3细胞因子含量
a,b,c:p<0.05。
实施例5
将健康的SPF级雄性大鼠(6-8周龄,200±20g),适应7天后,随机分为4组,每组10只。动物饲养保持环境温度为21±2℃,湿度为30-70%,12h光照交替,自由饮水,自由摄入饲料。基础饲料主要由鱼粉、小麦、玉米、豆粕、麸皮等组成;高脂高糖饲料是在基础饲料中添加15%蔗糖、15%猪油、10%酪蛋白。动物实验分组如下:
对照组:基础饲料喂养;
模型组:高脂高糖饲料喂养造模,使大鼠发生炎症;
实验组1:高脂高糖饲料喂养造模,同时灌胃本发明菌株悬液,灌胃剂量为1×109CFU/d;
实验组2:高脂高糖饲料喂养造模,同时灌胃本发明灭活菌株悬液,灌胃剂量为1×109CFU/d。
试验周期为10周,试验结束后,用1%戊巴比妥钠(0.5ml/100g BW)麻醉,采用心脏穿刺取血的方法获取大鼠血液样本,将血液样本取出后,静置30min,4000rpm,4℃离心15min,取上清,用ELISA试剂盒检测血清中IL-10、IL-1β、IL-6、TNF-α、MCP和NO含量。解剖取肝脏,用ELISA试剂盒检测肝脏中IL-1β、IL-6和NO含量。
由表4可知,与对照组相比,模型组大鼠血清中促炎因子IL-1β、IL-6、TNF-α和NO显著高于模型组(p<0.05,p<0.01),说明大鼠体内已经发生炎症。
由表4可知,与模型组相比,实验组1和实验组2大鼠血清中抑炎因子IL-10水平显著高于模型组(p<0.01);实验组1和实验组2大鼠血清中促炎因子IL-1β、IL-6、TNF-α、 MCP-1和NO显著低于模型组(p<0.01),结果与体外细胞实验结果一致。说明植物乳杆菌 1701活菌株或灭活菌株服用浓度在1×109CFU/d能够显著降低大鼠体内炎症水平,具有缓解慢性炎症的功能。
表4血清炎症相关细胞因子含量
处理 | 对照组 | 模型组 | 实验组1 | 实验组2 |
IL-10(ng/L) | 141.69±5.03** | 81.48±9.31 | 138.60±6.15** | 131.14±8.01** |
IL-1β(ng/L) | 71.02±3.58* | 125.49±4.41 | 73.29±4.84** | 70.86±5.46** |
IL-6(ng/L) | 91.50±3.14** | 163.36±5.52 | 54.79±4.05** | 68.47±4.29** |
TNF-α(ng/L) | 362.71±46.76** | 797.99±29.10 | 427.41±48.22** | 482.37±39.57** |
MCP-1(ng/L) | 102.40±4.55** | 127.08±8.92 | 77.18±1.55** | 93.82±2.92** |
NO(ng/μL) | 5.39±0.93* | 7.48±1.53 | 4.22±1.26** | 4.51±1.65** |
与模型组相比,*:p<0.05;**:p<0.01。
由表5可知,与对照组相比,模型组肝脏组织中促炎因子IL-6、IL-1β和NO显著高于对照组(p<0.05),说明肝脏已经发生炎症。
由表5可知,与模型组相比,实验组1和实验组2大鼠肝脏组织中促炎因子IL-6、IL-1β和NO显著低于模型组(p<0.01),与对照组相当。说明植物乳杆菌1701活菌株或灭活菌株服用浓度在1×109CFU/d能够显著降低大鼠肝脏组织炎症水平,具有缓解炎症的功能。
表5肝脏炎症相关细胞因子含量
处理 | IL-6(ng/L) | NO(ng/μL) | IL-1β(ng/L) |
对照组 | 231.63±10.51** | 0.51±0.06** | 16.90±1.78** |
模型组 | 353.71±12.69 | 0.95±0.08 | 29.19±1.51 |
实验组1 | 236.83±16.61** | 0.54±0.04** | 16.50±1.78** |
实验组2 | 235.43±11.24** | 0.64±0.05** | 16.29±1.20** |
与模型组相比,*:p<0.05;**:p<0.01。
综上所述,植物乳杆菌1701活菌株或灭活菌株服用浓度在1×109CFU/d能够综合调节体内多种炎症因子水平,使抑炎因子水平提高、促炎因子水平降低,从而缓解机体慢性炎症。
应用实施例
实施例1:植物乳杆菌1701冻干菌粉的制作
本发明菌株植物乳杆菌1701以1%的接种量接种于10mL液体MRS培养基中,在37℃恒温培养箱中培养16h(一代种子液)。一代种子液以1%的接种量接种于100mL液体MRS培养基中,在37℃恒温培养箱中培养16h(二代种子液)。二代种子1%的接种量接种于10L含有液体MRS培养基的发酵罐中,150rpm,pH 6.0,37℃培养16h,收集菌液,8000rpm离心10min收集菌体,用0.9%生理盐水洗涤一次,加入四倍菌泥量的含有脱脂乳粉、葡萄糖、甘油的保护剂中重悬,真空冷冻干燥,菌粉真空包装。制得的菌粉活菌数可达2×1011CFU/g,可用于含有植物乳杆菌1701的具有缓解慢性炎症功能相关药品、保健品、食品、饮料或发酵剂产品的制作和生产。
实施例2:植物乳杆菌1701发酵乳的制作
准确称取脱脂乳125g,45-50℃纯净水875g,50℃温水溶解,剪切20min,50℃水化30min,均质,95℃杀菌5min,降温后,按1%接种量接入应用实施例1中的植物乳杆菌1701发酵剂,37℃发酵12h,4℃后熟8-12h。观察其凝乳状态,用打蛋器破乳后,检测其产酸曲线、拉丝长度、pH、酸度、黏度和活菌数,设置三次重复。
结果如表6和图1所示,发酵乳凝乳状态紧实,有少量清洗出,破乳后,经较长时间搅拌丝滑粘稠,无拉丝,活菌数达1.56×108CFU/g,感官和风味良好,奶香浓郁,口感细腻丝滑,该发酵乳即为含有植物乳杆菌1701的具有缓解慢性炎症功能的发酵乳。
表6植物乳杆菌1701发酵乳特性
pH | 酸度(°T) | 黏度(cP) | 活菌数lg值(CFU/g) |
4.02±0.05 | 78.39±1.15 | 2127±13.20 | 8.19±0.13 |
本发明中所用原料、设备,若无特别说明,均为本领域的常用原料、设备;本发明中所用方法,若无特别说明,均为本领域的常规方法。
以上所述,仅是本发明的较佳实施例,并非对本发明作任何限制,凡是根据本发明技术实质对以上实施例所作的任何简单修改、变更以及等效变换,均仍属于本发明技术方案的保护范围。
序列表
<110> 杭州娃哈哈科技有限公司
<120> 一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用
<130> 2020
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1451
<212> DNA
<213> 植物乳杆菌1701(Lactobacillus plantarum 1701)
<400> 1
atacatgcag tcgaacgaac tctggtattg attggtgctt gcatcatgat ttacatttga 60
gtgagtggcg aactggtgag taacacgtgg gaaacctgcc cagaagcggg ggataacacc 120
tggaaacaga tgctaatacc gcataacaac ttggaccgca tggtccgagt ttgaaagatg 180
gcttcggcta tcacttttgg atggtcccgc ggcgtattag ctagatggtg aggtaacggc 240
tcaccatggc aatgatacgt agccgacctg agagggtaat cggccacatt gggactgaga 300
cacggcccaa actcctacgg gaggcagcag tagggaatct tccacaatgg acgaaagtct 360
gatggagcaa cgccgcgtga gtgaagaagg gtttcggctc gtaaaactct gttgttgaag 420
aagaacatat ctgagagtaa ctgttcaggt attgacggta tttaaccaga aagccacggc 480
taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gatttattgg 540
gcgtaaagcg agcgcaggcg gttttttaag tctgatgtga aagccttcgg ctcaaccgaa 600
gaagtgcatc ggaaactggg aaacttgagt gcagaagagg acagtggaac tccatgtgta 660
gcggtgaaat gcgtagatat atggaagaac accagtggcg aaggcggctg tctggtctgt 720
aactgacgct gaggctcgaa agtatgggta gcaaacagga ttagataccc tggtagtcca 780
taccgtaaac gatgaatgct aagtgttgga gggtttccgc ccttcagtgc tgcagctaac 840
gcattaagca ttccgcctgg ggagtacggc cgcaaggctg aaactcaaag gaattgacgg 900
gggcccgcac aagcggtgga gcatgtggtt taattcgaag ctacgcgaag aaccttacca 960
ggtcttgaca tactatgcaa atctaagaga ttagacgttc ccttcgggga catggataca 1020
ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080
cgcaaccctt attatcagtt gccagcatta agttgggcac tctggtgaga ctgccggtga 1140
caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200
cacgtgctac aatggatggt acaacgagtt gcgaactcgc gagagtaagc taatctctta 1260
aagccattct cagttcggat tgtaggctgc aactcgccta catgaagtcg gaatcgctag 1320
taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380
acaccatgag agtttgtaac acccaaagtc ggtggggtaa ccttttagga accagccgcc 1440
taagtggaca g 1451
Claims (8)
1.一株植物乳杆菌在制备缓解机体慢性炎症的组合物中的应用,其特征在于,所述组合物含有植物乳杆菌1701和/或其突变体;所述植物乳杆菌1701已在2019年10月23日保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号为CGMCC No.18728,微生物分类命名为植物乳杆菌Lactobacillus plantarum;所述突变体为对所述植物乳杆菌进行诱变、驯化、基因重组或者经自然突变而获得的突变体。
2.如权利要求1所述的应用,其特征在于,所述植物乳杆菌1701为活菌或死菌。
3.如权利要求1所述的应用,其特征在于,所述组合物还含有生理上可接受的赋形剂和/或稀释剂。
4.如权利要求3所述的应用,其特征在于,所述组合物为食品、药品、保健品或发酵剂。
5.如权利要求4所述的应用,其特征在于,所述食品为发酵乳、乳酪、含乳饮料、乳粉、固体饮料或发酵果蔬。
6.如权利要求4所述的应用,其特征在于,所述药品的制剂形式为胶囊、粉剂或片剂。
7.如权利要求4所述的应用,其特征在于,所述保健品的制剂形式为胶囊、粉剂或片剂。
8.如权利要求3所述的应用,其特征在于,所述组合物为口服形式。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113881604A (zh) * | 2021-11-10 | 2022-01-04 | 深圳大学 | 一株植物乳杆菌mm89及其多糖和应用 |
CN116042428A (zh) * | 2021-10-28 | 2023-05-02 | 南京盛德生物科技研究院有限公司 | 一种植物乳杆菌wsh044及其筛选方法及其发酵产物的制备方法 |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008110569A1 (en) * | 2007-03-12 | 2008-09-18 | Bioneer A/S | Method for determination of immunomodulatory effect |
WO2011118060A1 (ja) * | 2010-03-26 | 2011-09-29 | 株式会社明治 | 腸管免疫の抑制剤のスクリーニング方法 |
WO2012014971A1 (ja) * | 2010-07-30 | 2012-02-02 | 株式会社明治 | メタボリックシンドローム改善効果を有する乳酸菌 |
KR20130046897A (ko) * | 2011-10-28 | 2013-05-08 | 대상에프앤에프 주식회사 | 락토바실러스 플란타륨 dsr ck10또는 락토바실러스 플란타륨 dsr m2를 유효성분으로 함유하는 염증질환 치료용 조성물 |
CN107075455A (zh) * | 2014-08-22 | 2017-08-18 | 株式会社Atogen | 具有抗炎症及代谢疾病改善功效的植物乳杆菌hac01菌株及其用途 |
CN110055199A (zh) * | 2019-05-24 | 2019-07-26 | 吉林省命之元生物科技有限公司 | 一株植物乳杆菌ua149株及其应用 |
CN111019857A (zh) * | 2019-12-16 | 2020-04-17 | 吉林省命之元生物科技有限公司 | 一株植物乳杆菌hg20株及其应用 |
CN111254089A (zh) * | 2019-12-30 | 2020-06-09 | 杭州娃哈哈科技有限公司 | 一株具有减肥功能的植物乳杆菌及其应用 |
-
2020
- 2020-09-11 CN CN202010954814.4A patent/CN112190600B/zh active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008110569A1 (en) * | 2007-03-12 | 2008-09-18 | Bioneer A/S | Method for determination of immunomodulatory effect |
WO2011118060A1 (ja) * | 2010-03-26 | 2011-09-29 | 株式会社明治 | 腸管免疫の抑制剤のスクリーニング方法 |
WO2012014971A1 (ja) * | 2010-07-30 | 2012-02-02 | 株式会社明治 | メタボリックシンドローム改善効果を有する乳酸菌 |
KR20130046897A (ko) * | 2011-10-28 | 2013-05-08 | 대상에프앤에프 주식회사 | 락토바실러스 플란타륨 dsr ck10또는 락토바실러스 플란타륨 dsr m2를 유효성분으로 함유하는 염증질환 치료용 조성물 |
CN107075455A (zh) * | 2014-08-22 | 2017-08-18 | 株式会社Atogen | 具有抗炎症及代谢疾病改善功效的植物乳杆菌hac01菌株及其用途 |
CN110055199A (zh) * | 2019-05-24 | 2019-07-26 | 吉林省命之元生物科技有限公司 | 一株植物乳杆菌ua149株及其应用 |
CN111019857A (zh) * | 2019-12-16 | 2020-04-17 | 吉林省命之元生物科技有限公司 | 一株植物乳杆菌hg20株及其应用 |
CN111254089A (zh) * | 2019-12-30 | 2020-06-09 | 杭州娃哈哈科技有限公司 | 一株具有减肥功能的植物乳杆菌及其应用 |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116042428A (zh) * | 2021-10-28 | 2023-05-02 | 南京盛德生物科技研究院有限公司 | 一种植物乳杆菌wsh044及其筛选方法及其发酵产物的制备方法 |
CN113881604A (zh) * | 2021-11-10 | 2022-01-04 | 深圳大学 | 一株植物乳杆菌mm89及其多糖和应用 |
CN113881604B (zh) * | 2021-11-10 | 2023-02-28 | 深圳大学 | 一株植物乳杆菌mm89及其多糖和应用 |
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