高通量测序分析鞣花酸对大鼠肠道菌群的影响

2019-11-01 01:21沈起兵李淑娟刘同方张国文陈斌董海胜陈军丽于燕波
安徽农业科学 2019年18期
关键词:高通量测序肠道菌群

沈起兵 李淑娟 刘同方 张国文 陈斌 董海胜 陈军丽 于燕波

摘要[目的]采用高通量测序技术分析鞣花酸对大鼠肠道菌群组成和多样性的影响。[方法]采集持续灌胃鞣花酸SD大鼠的粪便,运用高通量测序技术对粪便肠道菌群进行分析,通过生物信息学方法分析大鼠肠道菌群的多样性和结构组成。[结果]通过高通量测序发现灌胃鞣花酸使SD大鼠肠道菌群的多样性增加(P<0.05),灌胃期间菌群的组成也发生了明显的波动,在门水平菌群变化中,拟杆菌门丰度增高(P<0.05),变形菌门丰度则降低到较低水平(P<0.01);在属水平菌群变化中,Bacteroidales S24-7 group、Lachnospiraceae NK4A136 group和拟杆菌属所占比例显著升高(P<0.05、0.01)。[结论]鞣花酸对大鼠肠道菌群具有调节作用。

关键词鞣花酸;肠道菌群;高通量测序

中图分类号R 965文献标识码A

文章编号0517-6611(2019)18-0171-03

doi:10.3969/j.issn.0517-6611.2019.18.046

开放科学(资源服务)标识码(OSID):

Effects of Ellagic Acid on Gut Microbiota in SD Rats Evaluated by High-throughput Sequencing

SHEN Qi-bing, LI Shu-juan, LIU Tong-fang et al

(SPAC Enter Space Science and Technology Institute, Shenzhen,Guangdong 518117)

Abstract[Objective]To analyze the effect of ellagic acid on the intestinal microbiota of SD rats by using Miseq high-throughput sequencing technology. [Method]SD rats were continuously fed with ellagic acid, the fecal samples during feeding were collected and analyzed by Illumina MiSeq for 16s sequencing. Afterwards, Bioinformatics methods were applied for illuminating diversity and constitution in gut microbita. [Result]The gut microflora had an upward trend in diversity (P<0.05) and remarkable variation in constitution. At the level of the phylum, the abundance of Bacteroides increased to a higher level (P<0.05), while that of Proteobacteria decreased gradually (P<0.01). At the genus level, the proportions of Bacteroidales S24-7 group, Lachnospiraceae NK4A136 group and Bacteroides were significantly elevated (P<0.05 or P<0.01). [Conclusion]Ellagic acid modulates the intestinal flora of SD rats.

Key wordsEllagic acid;Gut microbiota;High-throughput sequencing

鞣花酸,别名并没食子酸,广泛分布于石榴、板栗、草莓、黑莓、紅莓及胡桃等水果或坚果中。鞣花酸是富含羟基的天然多酚类化合物,其具有抗氧化、抗炎、抗肿瘤以及调节肠道菌群等多种生物活性,对预防或治疗慢性疾病如癌症、糖尿病和神经性病变等具有潜在的应用价值[1]。

动物肠道中栖息着数量巨大的微生物,统称为肠道菌群,其跟宿主的生理、代谢和免疫,甚至情绪和认知能力都有关联[2],肠道菌群一旦失调就会对宿主的健康产生影响。鞣花酸的抗氧化、抗炎及抗肿瘤活性已经被国内外学者系统论证,而其对肠道菌群影响的相关研究鲜有报道。笔者采用高通量测序技术考察鞣花酸对大鼠肠道菌群的影响,从改善肠道微生态环境的角度初步揭示鞣花酸调节肠道菌群的作用机制,为富含鞣花酸类食品资源的科学利用提供参考依据。

1材料与方法

1.1材料

SPF级SD大鼠6只,雄性,平均体质量为 (200±10)g,购自广东省医学动物实验中心,许可证号为SCXK(粤)2013-002。鞣花酸提取物,纯度为90%,购自陕西慧科植物开发有限公司。

1.2方法

1.2.1样本的采集与处理。

SD大鼠喂养于独立通风笼具中,每个笼具1只,光照黑暗时间比为12 h∶12 h,期间自由进食饮水,适应性喂养7 d后开始试验。采用自身前后对照,试验开始前收集空白粪便样本(Day 0)于干燥无菌的离心管中,试验开始后给予400 mg/kg的鞣花酸提取物,共28 d,分别在第7天(Day 7)、第14天(Day 14)、第21天(Day 21)、第28天(Day 28)采集粪便样本,采集后迅速密封,于 -80 ℃冰箱中冷冻保存。

參考文献

[1]SEERAM N P,SCHULMAN R N,HEBER D.Pomegranates:Ancient roots to modern medicine[M].Boca Raton:CRC Press,2006:244.

[2]BERCIK P,COLLINS S M,VERDU E F.Microbes and the gut-brain axis[J].Journal of neurogastroenterology and motility,2012,24(5):405-413.

[3]LI R Q,ZHU H M,RUAN J,et al.De novo assembly of human genomes with massively parallel short read sequencing[J].Genome research,2010,20(2):265-272.

[4]YANG F,ZENG X W,NING K,et al.Saliva microbiomes distinguish caries-active from healthy human populations[J].The ISME Journal,2012,6(1):1-10.

[5]赵菲,刘孝余,宫芳婧,等.高通量测序技术研究鱼腥草酵素对小鼠肠道菌群的影响[J].安徽农业科学,2018,46(32):156-159,172.

[6]李晓敏,杨丽杰,霍贵成.Illumina技术研究不同喂养方式婴儿肠道菌群差异[J].食品科技,2012,37(9):319-324.

[7]HATAYAMA H,WASHITA J,KUWAJIMA A,et al.The short chain fatty acid,butyrate,stimulates MUC2 mucin production in the human colon cancer cell line,LS174T[J].Biochemical and biophysical research communications,2007,356:599-603.

[8]LOUIS P,FLINT H J.Diversity,metabolism and microbial ecology of butyrate-producing bacteria from the human large intestine[J].FEMS Microbiology Letters,2009,294:1-8.

[9]WEXLER H M.Bacteroides:The good,the bad,and the nitty-gritty[J].Clinical microbiology reviews,2007,20(4):593-621.

[10]XU J,MAHOWALD M A,LEY R E,et al.Evolution of symbiotic bacteria in the distal human intestine[J].PLoS Biology,2007,5(7):1574-1586.

[11]MARTENS E C,CHIANG H C,GORDON J L.Mucosal glycan foraging enhances fitness and transmission of a saccharolytic human gut bacterial symbiont[J].Cell host microbe,2008,4(5):447-457.

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