目的

构建消化道术后吻合口漏小鼠动物模型。

方法

2023年6月至2023年12月将30只C57BL/6小鼠随机分为两组：对照组6只，仅做剖腹探查；实验组24只，从结肠无血管区剪破肠管并结扎供应此段的血管，然后进行吻合，记录手术时间。于术后2d行剖腹探查小鼠腹腔内部形态学变化，并采集吻合口周围组织，进一步观察吻合口漏组织学的变化。应用Image J和Graph Pad Prism 9.5软件进行数据统计分析。两组间比较采用独立样本 t检验，以 P＜0.05为差异有统计学意义。

结果

该模型构建平均时间为25min，小鼠的存活率为75%。亚甲蓝灌注实验显示肠管存在漏口。剖腹探查，肉眼可见吻合口存在轻微粘连，粘连部分可分离，肠管有漏口形成。与对照组比较，实验组苏木精-伊红染色可见吻合口有肌层不连续现象。Western Blot技术显示同一部位组织炎症因子在蛋白水平的表达差异明显。F4/80免疫组化显示在肌层断裂位置可见大量巨噬细胞浸润。

结论

通过该方法可成功构建消化道术后吻合口漏小鼠动物模型。

Objective

The mouse model of postoperative anastomotic leakage of digestive tract was established.

Methods

From June 2023 to December 2023, 30 C57BL/6 mice were randomly divided into two groups: 6 mice in the control group were performed laparotomy only; In the experimental group of 24, the intestinal tube was cut from the vascularized area of the colon and the blood vessels supplying this section were ligation, then anastomosis was performed, and the operation time was recorded.Two days after operation,a laparotomy was performed to investigate the morphological changes in the abdominal cavity of the mice, and the tissues around the anastomosis were collected to further observe the histological changes of the anastomotic leakage.Image J and Graph Pad Prism 9.5 software were used for statistical analysis.Independent sample t test was used for comparison between the two groups, and P＜0.05 was considered statistically significant.

Results

The average construction time of the model was 25min, and the survival rate of the mice was 75%.Methylene blue perfusion test showed leakage of intestinal tube.Laparotomy exploration showed that there was slight adhesion in the anastomosis, the adhesion part could be separated, and there was leakage in the intestinal tube.Compared with the control group, hematoxylin-eosin staining showed muscle discontinuity at the anastomosis in the experimental group.Western Blot showed that the expression of inflammatory factors in the same part of the tissue was significantly different at the protein level.F4/80 immunohistochemistry showed a large number of macrophage infiltrates at the site of muscle break.

Conclusion

Through this method, a mouse model of postoperative anastomotic leakage of digestive tract can be successfully constructed.