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小肠微创手术后结肠微生物群及肠道细胞因子基因表达的变化

Changes in the colon microbiota and intestinal cytokine gene expression following minimal intestinal surgery.

作者信息

Lapthorne Susan, Bines Julie E, Fouhy Fiona, Dellios Nicole L, Wilson Guineva, Thomas Sarah L, Scurr Michelle, Stanton Catherine, Cotter Paul D, Pereira-Fantini Prue M

机构信息

Susan Lapthorne, Nicole L Dellios, Sarah L Thomas, Michelle Scurr, Prue M Pereira-Fantini, Intestinal Failure and Clinical Nutrition group, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Victoria 3052, Australia.

出版信息

World J Gastroenterol. 2015 Apr 14;21(14):4150-8. doi: 10.3748/wjg.v21.i14.4150.

Abstract

AIM

To investigate the impact of minor abdominal surgery on the caecal microbial population and on markers of gut inflammation.

METHODS

Four week old piglets were randomly allocated to a no-surgery "control" group (n = 6) or a "transection surgery" group (n = 5). During the transection surgery procedure, a conventional midline incision of the lower abdominal wall was made and the small intestine was transected at a site 225 cm proximal to the ileocaecal valve, a 2 cm segment was removed and the intestine was re-anastomosed. Piglets received a polymeric infant formula diet throughout the study period and were sacrificed at two weeks post-surgery. Clinical outcomes including weight, stool consistency and presence of stool fat globules were monitored. High throughput DNA sequencing of colonic content was used to detect surgery-related disturbances in microbial composition at phylum, family and genus level. Diversity and richness estimates were calculated for the control and minor surgery groups. As disturbances in the gut microbial community are linked to inflammation we compared the gene expression of key inflammatory cytokines (TNF, IL1B, IL18, IL12, IL8, IL6 and IL10) in ileum, terminal ileum and colon mucosal extracts obtained from control and abdominal surgery groups at two weeks post-surgery.

RESULTS

Changes in the relative abundance of bacterial species at family and genus level were confined to bacterial members of the Proteobacteria and Bacteroidetes phyla. Family level compositional shifts included a reduction in the relative abundance of Enterobacteriaceae (22.95 ± 5.27 vs 2.07 ± 0.72, P < 0.01), Bacteroidaceae (2.54 ± 0.56 vs 0.86 ± 0.43, P < 0.05) and Rhodospirillaceae (0.40 ± 0.14 vs 0.00 ± 0.00, P < 0.05) following transection surgery. Similarly, at the genus level, changes associated with transection surgery were restricted to members of the Proteobacteria and Bacteroidetes phyla and included decreased relative abundance of Enterobacteriaceae (29.20 ± 6.74 vs 2.88 ± 1.08, P < 0.01), Alistipes (4.82 ± 1.73 vs 0.18 ± 0.13, P < 0.05) and Thalassospira (0.53 ± 0.19 vs 0.00 ± 0.00, P < 0.05). Surgery-associated microbial dysbiosis was accompanied by increased gene expression of markers of inflammation. Within the ileum IL6 expression was decreased (4.46 ± 1.60 vs 0.24 ± 0.06, P < 0.05) following transection surgery. In the terminal ileum, gene expression of TNF was decreased (1.51 ± 0.13 vs 0.80 ± 0.16, P < 0.01) and IL18 (1.21 ± 0.18 vs 2.13 ± 0.24, P < 0.01), IL12 (1.04 ± 0.16 vs 1.82 ± 0.32, P < 0.05) and IL10 (1.04 ± 0.06 vs 1.43 ± 0.09, P < 0.01) gene expression increased following transection surgery. Within the colon, IL12 (0.72 ± 0.13 vs 1.78 ± 0.28, P < 0.01) and IL10 (0.98 ± 0.02 vs 1.95 ± 0.14, P < 0.01) gene expression were increased following transection surgery.

CONCLUSION

This study suggests that minor abdominal surgery in infants, results in long-term alteration of the colonic microbial composition and persistent gastrointestinal inflammation.

摘要

目的

研究小型腹部手术对盲肠微生物群落及肠道炎症标志物的影响。

方法

将四周龄仔猪随机分为非手术“对照组”(n = 6)和“横断手术组”(n = 5)。在横断手术过程中,在下腹部壁做常规中线切口,在距回盲瓣近端225 cm处横断小肠,切除2 cm肠段,然后将肠管重新吻合。在整个研究期间,仔猪接受聚合型婴儿配方奶粉喂养,并在术后两周处死。监测包括体重、粪便稠度和粪便脂肪球存在情况等临床结果。采用高通量DNA测序分析结肠内容物,以检测门、科和属水平上与手术相关的微生物组成紊乱情况。计算对照组和小型手术组的多样性和丰富度估计值。由于肠道微生物群落紊乱与炎症有关,我们比较了术后两周从对照组和腹部手术组获得的回肠、回肠末端和结肠黏膜提取物中关键炎症细胞因子(TNF、IL1B、IL18、IL12、IL8、IL6和IL10)的基因表达。

结果

科和属水平上细菌种类相对丰度的变化仅限于变形菌门和拟杆菌门的细菌成员。科水平的组成变化包括横断手术后肠杆菌科(22.95±5.27 vs 2.07±0.72,P<0.01)、拟杆菌科(2.54±0.56 vs 0.86±0.43,P<0.05)和红螺菌科(0.40±0.14 vs 0.00±0.00,P<0.05)相对丰度降低。同样,在属水平上,与横断手术相关的变化仅限于变形菌门和拟杆菌门的成员,包括肠杆菌科(29.20±6.74 vs 2.88±1.08,P<0.01)、艾氏菌属(4.82±1.73 vs 0.18±0.13,P<0.05)和海螺旋菌属(0.53±0.19 vs 0.00±0.00,P<0.05)相对丰度降低。手术相关的微生物失调伴随着炎症标志物基因表达的增加。在回肠内,横断手术后IL6表达降低(4.46±1.60 vs 0.24±0.06,P<0.05)。在回肠末端,横断手术后TNF基因表达降低(1.51±0.13 vs 0.80±0.16,P<0.01),IL18(1.21±0.18 vs 2.13±0.24,P<0.01)、IL12(1.04±0.16 vs 1.82±0.32,P<0.05)和IL10(1.04±0.06 vs 1.43±0.09,P<0.01)基因表达增加。在结肠内,横断手术后IL12(0.72±0.13 vs 1.78±0.28,P<0.01)和IL10(0.98±0.02 vs 1.95±0.14,P<0.01)基因表达增加。

结论

本研究表明,婴儿小型腹部手术会导致结肠微生物组成的长期改变和持续的胃肠道炎症。

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