| Objective:To reduce surgical stress and improve experimental repeatability,using minimally invasive surgery establish an animal mode.At present,the method of establishing canine left liver lobectomy model with minimally invasive technique is complicated,expensive and hard to popularize.In this study,we created a new method for the left hepatectomy in Beagles,as well as monitored the safety and liver injury of the model and analyzed the gut microbiota changes of canines after operation.Methods:A purely laparoscopic left liver lobectomy(left medial lobe and left lateral lobe)was performed by four-port in 6 beagles.First,the left triangular ligament was removed and the hepatic pedicle of the left inner lobe was ligated with needle suture.Then,the left medial liver lobe was removed using LigaSure about 0.5 cm above the ligation site,and the left lateral liver lobe was removed in the same way.Finally,a cannula hole is enlarged to retrieve the liver lobe in the bag.To evaluate the safety of the model,the changes of animal physiological indexes,complete blood count and serum organics or enzyme analyses were monitored.And the degree of liver injury was verified by the ratio of CD3+-CD4+/CD3+-CD8+ in peripheral blood,inflammatory factors and liver histopathology.Beagles' feces were collected during perioperative period.After extracting the total DNA,PCR amplification was performed on the V3-V4 region of 16s rDNA of microorganisms,and the amplified products were sequenced.Results:Six Beagles were successfully removed from the left medial lobe and left lateral lobe,with perioperative stable physiological indexes and rapid recovery after operation without complications.WBC and NEUT were increased 1 day after surgery,and the changes were extremely significant differences compared with those before surgery(p<0.01).ALT,AST and ALP were increased 1 d and 3 d after surgery,and were significant differences compared with those before surgery(p<0.05),and then gradually recovered.Proinflammatory factors increased(IL-??IL-6?TNF-?)and anti-inflammatory factors decreased(IL-4?IL-10)3 d after the operation.The changes of inflammatory factors 1 d and 3 d after the operation were extremely significantly differences compared with those before the operation(p<0.01),and then gradually recovered.Histopathology showed severe liver injury and immune function was not significantly changes.After high-throughput sequencing of 16s rDNA,the diversity of gut microbiota was significantly difference.At the level of phylum,Firmicutes were always the dominant flora during perioperative period and Proteobacteria increased 7 d after the operation,with extremely significantly changes compared with those before the operation(p<0.01).The relative abundance of Peptoclostridium,Allobaculum,Turicibacter,Escherichia Shigella,and Erysipelotrichaceae_norank were among the top ten populations at 7 days after surgery on the genus level,and were significantly differences from those before surgery(p<0.05).At the genus level,LEfse analysis and correlation analysis showed that Allobaculum,Turicibacter and Escherichia Shigella were closely related to the change of inflammation and liver function after hepatectomy.PICRUSt analysis showed that the relative abundance of 10 KEGG pathways was significant difference after hepatectomy(p<0.05).Conclusions:(1)In this study,the beagle model of liver lobectomy was successfully established by laparoscopic hepatopedicle ligation combined with parenchymal resection by LigaSure.The method is safe and effective,and can be applied to liver regeneration,liver surgery,liver protective drugs and other studies.(2)Hepatectomy had a serious impact on gut microbiota.Allobaculum,Turicibacter and Escherichia Shigella had significant changes as well as closely related to inflammatory response and liver injury after surgery,which could be used as potential intervention targets after liver surgery. |
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