CN113018319A - 一种用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的复合菌剂及其应用 - Google Patents
一种用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的复合菌剂及其应用 Download PDFInfo
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- CN113018319A CN113018319A CN202110080742.XA CN202110080742A CN113018319A CN 113018319 A CN113018319 A CN 113018319A CN 202110080742 A CN202110080742 A CN 202110080742A CN 113018319 A CN113018319 A CN 113018319A
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Abstract
本发明提供一种用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的复合菌剂及其应用,所述复合菌剂的活性组分包括益生菌组合物和功能性物质,所述益生菌组合物包括鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌,所述功能性物质包括瓜氨酸、谷胱甘肽、辅酶Q10和沙棘提取物中的至少一种,在所述益生菌组合物中,总活菌数≥5.0×108CFU/g,鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥8.0×107CFU/g,动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥3.5×107CFU/g。本发明提供的复合菌剂可缓解因长期低剂量辐射暴露所导致的动物胰岛素抵抗。
Description
技术领域
本发明涉及生物医药技术领域,特别地,涉及一种用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的复合菌剂,以及该复合菌剂的具体应用。
背景技术
国际原子能组织规定,不超过100mSv的电离辐射为低剂量辐射。而根据辐射流行病学以及临床研究表明,低剂量辐射可诱发多种疾病,比如癌症、先天异常、心脑血管疾病、认知及其他神经系统紊乱、白内障、内分泌紊乱、支气管肺损伤、消化系统损伤等其他功能损伤。相比于高剂量辐射,人类更容易被暴露在低剂量辐射环境下,例如核工业工人、放射性研究工作者、航空飞行员等,目前已有关于这些职业因辐射暴露致癌的报道。
此外,近期研究报道发现辐射对胰岛素抵抗有一定诱发作用。胰岛素抵抗是指各种原因使胰岛素促进葡萄糖摄取和利用的效率下降,机体代偿性的分泌过多胰岛素产生高胰岛素血症,以维持血糖的稳定。胰岛素抵抗容易引发代谢综合征和2型糖尿病。在一个小鼠试验中发现,在对小鼠进行全身照射后的第三天,辐射模型组的小鼠空腹血糖显著增加,且辐射小鼠胰腺氧化还原失衡、血清胰岛素水平降低、HOMA-β值降低,胰腺组织形态学观察结果都直接和间接显示辐射诱导氧化应激所致的胰腺细胞结构和功能的氧化损伤。同时,其他研究还表明,辐射能通过改变细胞周期调节因子,造成胰腺β细胞增殖分化失调;能通过调节胰岛素途径调控糖异生;还可以通过造成肝细胞内质网的氧化损伤等途径来影响糖异生,从而导致以持续性血糖升高为主要特点的糖代谢异常。
目前,对于长期低剂量照射导致机体出现胰岛素抵抗的问题尚无有效的干预办法。
发明内容
本发明的目的在于提供一种可缓解由长期慢性低剂量辐射所致胰岛素抵抗的技术方案,以解决上述问题。本发明技术方案如下。
一种复合菌剂在制备用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的产品过程中的应用,所述复合菌剂的活性组分包括益生菌组合物,所述益生菌组合物包括鼠李糖乳杆菌和嗜酸乳杆菌中的至少一种。
优选地,所述益生菌组合物还包括动物双歧杆菌和长双歧杆菌中的至少一种。
优选地,所述益生菌组合物包括鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌。
优选地,所述益生菌组合物中的总活菌数≥5.0×108CFU/g,其中,鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥8.0×107CFU/g,且动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥3.5×107CFU/g。
优选地,所述复合菌剂还包括功能性物质,所述功能性物质包括瓜氨酸、谷胱甘肽、辅酶Q10和沙棘提取物中的至少一种。
优选地,在所述复合菌剂中,益生菌组合物的含量为70-80%。
优选地,所述嗜酸乳杆菌具体为罗伊氏乳杆菌。
优选地,所述产品中包含所述复合菌剂。
优选地,所述产品为食品、保健品、特医食品和药品中的任意一种。
优选地,所述食品为固体饮料、酸奶、益生菌饮料和酸奶中的任意一种。
本发明提供的技术方案至少具有如下有益效果:
本发明通过提供一种包含益生菌组合物的复合菌剂,实现对肠道菌群生态的调节,该益生菌组合物可以显著提升经长期低剂量辐射后机体肠道内有益菌群的数量,改善肠道粘附液粘附能力,降低血清中炎症因子的表达水平,从而缓解胰岛素抵抗程度。
本发明提供的复合菌剂在缓解低剂量长期慢性辐射对胰岛素抵抗的影响方面具有很好的应用前景,且应用方式多样,治疗成本低,是一种安全、无毒副作用的提升亚健康人群的健康状况的方式
保藏说明
鼠李糖乳杆菌(Lactobacillus rhamnosus),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址:北京市朝阳区北辰西路1号院3号,保藏机构:CGMCC,保藏日期:2020年04月23日,生物保藏编号:CGMCC No.19711,菌株编号:KLrh-10。
动物双歧杆菌(Bifidobacterium animalis),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址:北京市朝阳区北辰西路1号院3号,保藏机构简称:CGMCC,保藏日期:2020年04月23日,生物保藏编号:CGMCC No.19712,菌株编号:KBla-22。
附图说明
为了更清楚地说明本发明实施例中的技术方案,下面将对实施例描述中所需要使用的附图作简单地介绍,显而易见地,下面描述的附图仅仅是本发明的一些实施例,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它的附图,其中:
图1是小鼠肠道菌群种水平受低剂量辐射及益生菌干预后的对比图;
图2是小鼠胰腺细胞受低剂量辐射及益生菌干预后氧化损伤的对比图;
图3是小鼠体内总胆固醇含量受低剂量辐射及益生菌干预后的对比图;
图4是小鼠体内低密度脂蛋白含量受低剂量辐射及益生菌干预后的对比图;
图5是小鼠体内高密度脂蛋白含量受低剂量辐射及益生菌干预后的对比图;
图6是小鼠体内空腹胰岛素抵抗水平受低剂量辐射及益生菌干预后的对比图。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅是本发明的一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其它实施例,都属于本发明保护的范围。
实施例1复合菌剂的制备
一种用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的复合菌剂,包括益生菌组合物和功能性物质。所述益生菌组合物包括鼠李糖乳杆菌、罗伊氏乳杆菌、动物双歧杆菌和长双歧杆菌。所述功能性物质包括瓜氨酸、谷胱甘肽、辅酶Q10和沙棘提取物。
所述益生菌组合物中的总活菌数≥5.0×108CFU/g,其中,鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥8.0×107CFU/g,且动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥3.5×107CFU/g。
所述益生菌组合物的制备过程为:
1)制作发酵培养基
每升发酵培养基按质量份包括如下组分:葡萄糖1-10份、蔗糖1-8份、麸皮2-15份、蛋白胨1-5份、椰子粉3-7份、脱脂大豆粉5-10份,纯化水100-400份;
2)制作各益生菌对应的液体菌种
将活化好的鼠李糖乳杆菌接种于装有100ml发酵培养基的500ml三角瓶中,在37℃环境下振荡培养48h,振荡速率为1000rpm且旋转半径为20mm,收集三角瓶内的所有培养物,即得到鼠李糖乳杆菌的液体菌种;
将活化好的罗伊氏乳杆菌接种于装有100ml发酵培养基的500ml三角瓶中,在37℃环境下振荡培养48h,振荡速率为1000rpm且旋转半径为20mm,收集三角瓶内的所有培养物,即得到罗伊氏乳杆菌的液体菌种;
将活化好的动物双歧杆菌接种于装有100ml发酵培养基的500ml三角瓶中,在37℃环境下振荡培养48h,振荡速率为1000rpm且旋转半径为20mm,收集三角瓶内的所有培养物,即得到动物双歧杆菌的液体菌种;
将活化好的长双歧杆菌接种于装有100ml发酵培养基的500ml三角瓶中,在37℃环境下振荡培养48h,振荡速率为1000rpm且旋转半径为20mm,收集三角瓶内的所有培养物,即得到长双歧杆菌的液体菌种;
3)将制得的鼠李糖乳杆菌、罗伊氏乳杆菌、动物双歧杆菌和长双歧杆菌的液体菌种分别稀释至8.0×107CFU/g,并依次按照1:4:3:2的体积比混合,即得到目标益生菌组合物。
将上述益生菌组合物与功能性物质混合得到目标复合菌剂,其中,益生菌组合物的含量为75%,功能性物质的含量为25%。
功效性物质的添加量参照了相应的国家标准,如GB 2760-2014《食品添加剂使用标准》和GB 14880-2012《食品营养强化剂使用标准》,相应的行业标准以及由国家卫生部等主管部门颁布的最新新食品原料使用规范,配料添加量均符合相应的法规规定。
实施例2小鼠辐射实验
本实施例中的动物实验均按照国家《实验动物管理条例》相关规定进行,并得到军事医学院实验动物管理与动物福利伦理委员会允许。
实验对象为6-8周龄的C57BL/6J雄性小鼠,每只小鼠的体重为20-25g,共计30只,每5只小鼠一笼。将上述30只小鼠分为对照组、低剂量辐射组和益生菌干预组,每组各10只。
对于对照组,小鼠采用正常饲养,动物房昼夜交替12h/12h,室温20-24℃,湿度(40%-60%)的负压环境中,自由饮食。
对于低剂量辐射组,在正常饲养的基础上,以单次0.05Gy钴-60射线辐照,一星期照射两次,共计20次。
对于益生菌干预组,在正常饲养的基础上,以单次0.05Gy钴-60射线辐照,一星期照射两次,共计20次,且在饲喂小鼠时往饲料中添加复合菌剂发酵粉,所述复合菌剂发酵粉是通过将复合菌剂在发酵培养物中培养48小时,然后在发酵液中加入多孔淀粉,混合均匀后冷冻干燥制成。
在照射结束后30天,分别收集3组小鼠肛门口的新鲜粪便,样品置于预冷的无菌EP管中,随后提取基因组,进行肠道菌群16rS测定分析。此外,还收集小鼠的血液样本,进行相应的检测分析。具体检测结果参见附图1-6和表1。
参见图1,CTR表示对照组,Treat A表示低剂量辐射组,Treat B表示益生菌干预组。其中:
Firmicutes:厚壁菌,是一大类细菌,多数为革兰氏阳性,肠内厚壁菌门多于拟杆菌门导致更有效吸收食物中的热量从而导致肥胖。
Bacteroidetes:拟杆菌门,包括拟杆菌纲(Bacteroidia)、黄杆菌纲(Flavobacteriia)和鞘脂杆菌纲(Sphingobacteriia),拟杆菌门为产甲烷过程的优势群落。
Others:包括Proteobacteria、unidentified_Bacteria、Tenericutes、Melainabacteria、Actinobacteria、Verrucomicrobia、Cyanobacteria、Euryarchaeota等其他肠道菌种。
表1小鼠肠道菌群种水平受低剂量辐射及益生菌干预后的对比
如图1和表1所示,在种水平上,与对照组相比,低剂量照射组中小鼠肠道内有害菌属Firmicutes和Proteobacteria等的表达丰度发生增高,而Bacteroidetes和Tenericutes的表达丰度发生降低;益生菌干预后,与低剂量辐射组相比,小鼠肠道内的Bacteroidetes和Melainabacteria的表达丰度发生增高,Firmicutes、unidentified_Bacteria和Proteobacteria等的表达丰度发生降低。
参见图2,其中,图2中的(a)是对照组的胰腺细胞照片,图2中的(b)是低剂量辐射组的胰腺细胞照片,图2中的(c)是益生菌干预组的胰腺细胞照片。
胰腺是分泌胰岛素的器官,胰腺损伤会造成胰岛素分泌障碍,使得机体胰岛素功能发生紊乱,引起胰岛素抵抗和糖尿病发生发展。如图2所示,与对照组相比,低剂量辐射组中小鼠的胰腺细胞发生明显的氧化损伤,而采用益生菌干预后,与低剂量辐射组相比,小鼠胰腺细胞的氧化损伤程度明显降低。
总胆固醇含量是一种可反映胰岛素抵抗严重程度的血脂指标,因为胰岛素抵抗会引起血脂代谢紊乱,其具体表现为总胆固醇含量升高。如图3所示,与对照组相比,低剂量辐射组中小鼠体内的总胆固醇含量明显增多,而用益生菌干预后,与低剂量辐射组相比,小鼠体内的总胆固醇含量显著降低。
低密度脂蛋白是一种可反映胰岛素抵抗严重程度的血脂指标,研究表明低密度脂蛋白是胰岛素抵抗的重要危险因子,能促进胰岛素抵抗的发生和发展。如图4所示,与对照组相比,低剂量辐射组中小鼠体内的低密度脂蛋白含量明显增多,而采用益生菌干预后,与低剂量辐射组相比,小鼠体内的低密度脂蛋白含量显著降低。
高密度脂蛋白是心脑血管的一种保护性蛋白,其含量降低是胰岛素抵抗的重要危险因子,胰岛素抵抗会引起血脂代谢紊乱,其具体表现为高密度脂蛋白含量降低。如图5所示,与对照组相比,低剂量辐射组中小鼠体内的高密度脂蛋白含量明显减少,而采用益生菌干预后,与低剂量辐射组相比,小鼠体内的高密度脂蛋白含量显著增多。
如图6所示,与对照组相比,低剂量辐射组中小鼠体内的空腹胰岛素抵抗水平明显升高,而采用益生菌干预后,与低剂量辐射组相比,小鼠体内的空腹胰岛素抵抗水平显著降低。
以上研究证明,通过本发明提供的复合菌剂调整菌群,重建肠道微生态,可显著提升肠道内有益菌群的数量,降低血清中炎症因子的表达水平,进而缓解长期低剂量辐射暴露所致胰岛素抵抗带来的影响。
以上所述仅为本发明的优选实施例,并非因此限制本发明的专利保护范围,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。在本发明的精神和原则之内,凡是利用本发明说明书及附图内容所作的任何改进或等同替换,直接或间接运用在其它相关的技术领域,均应包括在本发明的专利保护范围内。
Claims (9)
1.一种复合菌剂在制备用于缓解因长期低剂量辐射暴露所致胰岛素抵抗的产品过程中的应用,其特征在于,所述复合菌剂的活性组分包括益生菌组合物,所述益生菌组合物包括鼠李糖乳杆菌和嗜酸乳杆菌中的至少一种。
2.根据权利要求1所述的应用,其特征在于,所述益生菌组合物还包括动物双歧杆菌和长双歧杆菌中的至少一种。
3.根据权利要求2所述的应用,其特征在于,所述益生菌组合物包括鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌。
4.根据权利要求3所述的应用,其特征在于,所述益生菌组合物中的总活菌数≥5.0×108CFU/g,其中,鼠李糖乳杆菌、嗜酸乳杆菌、动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥8.0×107CFU/g,且动物双歧杆菌和长双歧杆菌中的单一益生菌的活菌数≥3.5×107CFU/g。
5.根据权利要求1-4中任意一项所述的应用,其特征在于,所述复合菌剂还包括功能性物质,所述功能性物质包括瓜氨酸、谷胱甘肽、辅酶Q10和沙棘提取物中的至少一种。
6.根据权利要求5所述的应用,其特征在于,在所述复合菌剂中,益生菌组合物的含量为70-80%。
7.根据权利要求6所述的应用,其特征在于,所述嗜酸乳杆菌具体为罗伊氏乳杆菌。
8.一种根据权利要求1-7中任意一项所述应用制备得到的产品,其特征在于,所述产品中包含所述复合菌剂。
9.根据权利要求8所述的产品,其特征在于,所述产品为食品、保健品、特医食品和药品中的任意一种。
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