基于高通量测序技术分析加速康复外科管理流程对结直肠癌患者肠道菌群的影响

Influence of enhanced recovery after surgery on intestinal flora of patients with colorectal cancer based on high-throughput sequencing technology

目的探讨加速康复外科(ERAS)管理流程对结直肠癌患者肠道菌群的影响。方法采用便利抽样方法纳入2018年8月至2019年12月期间四川大学华西医院胃肠外科收治的结直肠癌患者为研究对象,被随机分为ERAS组和传统组,其中ERAS组患者围术期管理依据ERAS流程进行,传统组按照传统流程进行围术期临床管理。于入院24 h内、术后第1次自然排便时分别采集研究对象的新鲜粪便标本,提取其总DNA,并对16 Sr DNA V3~V4区进行扩增和高通量测序,基于OTU聚类结果进行生物信息学分析。结果本研究共纳入60例结直肠癌患者,传统组30例,ERAS组27例(3人临时退出研究),2组患者的基本情况比较差异无统计学意义(P>0.05)。生物信息学分析结果:①alpha多样性的Shannon指数和Simpson指数在术前或术后的ERAS组与传统组间比较差异均无统计学意义(P>0.05),其在同一组内的术前与术后比较差异亦均无统计学意义(P>0.05)。②beta多样性分析结果显示,术前传统组和ERAS组的群落构成比较差异不明显,术后传统组和ERAS组的群落构成出现明显分界。③进一步对群落结构分布结果显示,门水平下,与术前丰度比较,厚壁菌门在传统组术后下降26.5%、ERAS组仅下降5.5%,拟杆菌门在传统组术后提升21.6%、ERAS组仅提升4.7%,变形菌门在传统组术后提升7.2%、ERAS组仅提升2.2%;属水平下,与术前丰度比较,拟杆菌属在传统组术后提升17.6%、ERAS下降1.6%,而双歧杆菌属在传统组术后下降1.8%、ERAS组下降1.3%。结论ERAS流程并不会对肠道菌群的物种多样性造成影响,其对于肠道菌群结构有一定的损伤,但较传统流程的损伤程度更弱,更有利于肠道微生态环境的重建和恢复。

Objective To explore the influence of enhanced recovery after surgery(ERAS)on intestinal flora in patients with colorectal cancer.Methods By convenient sampling method,60 patients with colorectal cancer were selected from August 2018 to December 2019 in the Department of Gastrointestinal Surgery of West China Hospital of Sichuan University and randomly divided into ERAS group and traditional treatment group(traditional group).Among them,the perioperative clinical management was carried out according to the ERAS management and traditional treatment process in the the ERAS group and in the traditional group,respectively.The fresh fecal samples were collected within 24 h after admission and the first natural defecation after operation.The bacterial 16 Sr DNA V3-V4 region was sequenced by Illumina MiSeq sequencer,and the results were analyzed by bioinformatics.Results A total of 60 patients with colorectal cancer were included,30 cases in the traditional group and 27 cases in the ERAS group(3 people temporarily withdrew from the study).There were no significant differences in the basic informations between the two groups(P>0.05).①Before or after operation,there were no significant differences in Shannon index and Simpson index between the two groups.The difference between preoperative and postoperative comparison in the same group was also not statistically significant(P>0.05).②Beta diversity analysis showed that there was no significant difference in community composition between the traditional group and the ERAS group before operation,and there was a clear boundary between the traditional group and the ERAS group after operation.③At the phylum level,compared with the preoperative abundance,the postoperative abundance Firmicutes decreased by 26.5%and 5.5%in the traditional and ERAS group,respectively;Bacteroidetes increased by 21.6%and 4.7%in the traditional and ERAS group,respectively;Proteobacteria increased by 7.2%and 2.2%in the traditional and ERAS group,respectively.At the genus level,compared with the preoperative abundance,the postoperative abundance of Bacteroides in the traditional group increased by 17.6%and in the ERAS group decreased by 1.6%;Bifidobacterium decreased by 1.8%and 1.3%in the traditional group and in the ERAS group,respectively.Conclusions ERAS does not affect species diversity of intestinal flora.Although ERAS has some damage to structure of intestinal flora,it is weaker than traditional process,so it is more conducive to reconstruction and restoration of intestinal microecological environment.