Shi Shang, Liu Yang, Wang Zhiyue, Jin Xiangren, Yan Wei, Guo Xiao, Lin Baiqiang, Wang Haoran, Li Bowen, Zheng Jianjun, Wei Yunwei
Oncology and Laparoscopy Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Pancreatic and Gastrointestinal Surgery Division, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo, China.
Front Microbiol. 2022 Dec 14;13:1031882. doi: 10.3389/fmicb.2022.1031882. eCollection 2022.
Despite advances in anastomotic techniques and perioperative care, the incidence of anastomotic leak (AL) has not substantially decreased over time. Although it is known that AL etiology is multifactorial and the mechanisms involved remain unclear, there is accumulating evidence pointing at AL related to gut microbiota.
We firstly performed a clinical study to analyze the gut microbiota between colorectal cancer patients who developed AL and those who did not (nAL) using 16S-rRNA sequencing and quantitative real-time PCR to identify AL risk bacterial taxa. Then we built a rat anastomosis model and performed a bacteria transplantation to ensure the cause-effect relationship. The anastomotic healing score was used to evaluate the healing of anastomosis. In addition, we assessed the adhesion ability of bacteria by staining with fluorescein isothiocyanate and attachment assay. The expression of matrix metalloproteinase 9 (MMP9) was detected by western blot, and the activity was detected by gelatin zymography.
We found that the abundance and positive rate of () were higher in the AL patients. Exposure of the rat's colon anastomosis to contributes to the loss of submucosa collagen I and III, leading to AL's pathogenesis. can attach to the gut epithelial cells and stimulate intestinal MMP9 expression and . We further confirmed that these effects of depended on the E-cadherin/β-catenin signaling pathway.
This work demonstrates that attaches and then stimulates the expression of epithelial cells MMP9 by the E-cadherin/β-catenin signaling pathway. These effects contribute to collagen break down in the intestinal tissue, finally leading to AL.
尽管吻合技术和围手术期护理取得了进展，但吻合口漏（AL）的发生率并未随时间显著降低。虽然已知AL的病因是多因素的，且相关机制仍不清楚，但越来越多的证据指向与肠道微生物群相关的AL。
我们首先进行了一项临床研究，使用16S-rRNA测序和定量实时PCR分析发生AL的结直肠癌患者与未发生AL的患者（nAL）之间的肠道微生物群，以确定AL风险细菌分类群。然后我们建立了大鼠吻合模型并进行细菌移植以确保因果关系。吻合口愈合评分用于评估吻合口的愈合情况。此外，我们通过异硫氰酸荧光素染色和附着试验评估细菌的黏附能力。通过蛋白质印迹法检测基质金属蛋白酶9（MMP9）的表达，并通过明胶酶谱法检测其活性。
我们发现，（此处原文缺失具体细菌名称）在AL患者中的丰度和阳性率更高。大鼠结肠吻合口暴露于（此处原文缺失具体细菌名称）会导致黏膜下层I型和III型胶原蛋白丢失，从而导致AL的发病机制。（此处原文缺失具体细菌名称）可以附着于肠道上皮细胞并刺激肠道MMP9表达（此处原文缺失具体内容）。我们进一步证实，（此处原文缺失具体细菌名称）的这些作用依赖于E-钙黏蛋白/β-连环蛋白信号通路。
这项工作表明，（此处原文缺失具体细菌名称）附着后通过E-钙黏蛋白/β-连环蛋白信号通路刺激上皮细胞MMP9的表达。这些作用导致肠道组织中的胶原蛋白分解，最终导致AL。
