Two-step Hepatectomy Combined With Liver Resection And Portal Vein Ligation And Clinical And Basic Research On Liver Fibrosis And AQP9 In Liver Regeneratio

Background:In China,associating liver partition and portal vein ligation for staged hepatectomy(ALPPS)is mainly used in primary liver cancer.Patients with primary liver cancer often have liver fibrosis,and the incidence of complications and mortality after ALPPS is high.Therefore,the application of ALPPS is controversial.Methods:This study reviewed the clinical data of patients with primary liver cancer who underwent ALPPS in Peking Union Medical College Hospital from May 2014 to October 2022.The degree of liver fibrosis was graded by hematoxylin-eosin staining and Sirius red staining.The effects of liver fibrosis on liver regeneration rate,postoperative complications and prognosis after ALPPS were analyzed.Results:Thirty patients with primary liver cancer who underwent ALPPS were included,including 23 patients with hepatocellular carcinoma,5 patients with cholangiocarcinoma,and 2 patients with combined hepatocellular-cholangiocarcinoma.There were 14 patients in the severe liver fibrosis group and 16 patients in the non-severe liver fibrosis group.All patients underwent two-step hepatectomy.The median interval between the two steps of ALPPS was 12.35 days(interquartile range IQR 8.50,20.29).The residual liver volume increased by 202.5 ml(IQR 104,269).The effect of severe liver fibrosis on liver regeneration rate was not statistically significant(P=0.892).The incidence of severe complications after the first step of ALPPS was 6.67%,mainly transient renal failure,while that after the second step was 13.33%,mainly liver failure.Patients with severe complications all belonged to severe liver fibrosis group.The mortality was 6.67%in the 30 days and 16.67%in 90 days after the operation.Severe liver fibrosis was significantly associated with 90-day mortality(P=0.014),and overall survival(P=0.012).Conclusion:Severe liver fibrosis has little effect on the rate of liver regeneration after the first step of ALPPS,and severe liver fibrosis is an important risk factor for liver failure and perioperative death after the second step of ALPPS.Preoperative liver function impairment is an important predictive factor for postoperative liver failure.Background:It is controversial whether associating liver partition and portal vein ligation for staged hepatectomy(ALPPS)can promote liver regeneration in rats with liver fibrosis.In this study,the first step of ALPPS model with liver fibrosis was established by using carbon tetrachloride(CCl4)and ethanol to further clarify the effect of liver fibrosis on liver regeneration after the first step of ALPPS in rats.Methods:Totally 40 male SD rats were randomly divided into two groups with 20 rats in each group.The control group was subcutaneously injected with olive oil and routinely fed for 4 weeks,and the liver fibrosis group was subcutaneously injected with CCl4 and forcedly fed with 10%ethanol for 4 weeks.Sirius red staining was used to judge the degree of liver fibrosis.The regenerated liver was weighed immediately,3 days and 6 days after the first step of ALPPS in rats,and the regeneration of hepatocytes was further judged by Ki67 immunohistochemical staining.Results:After 4 weeks of CCl4 and ethanol modeling,the weight of rats increased slowly,their life was basically unaffected,and liver fibrosis was uniform.One rat died due to the injection of CCl4,and the success rate of liver fibrosis modeling was 95.0%.The rats in the liver fibrosis group and the control group all survived after the first step of ALPPS,and the success rate of ALPPS modeling was 100%.Both groups achieved rapid growth of future liver volume,and there was no significant difference between the two groups(P>0.05).Ki67 positive cells in the hepatic fibrosis group and the control group increased significantly on the 3rd day after the operation and decreased on the 6th day after the operation.Conclusion:Using the method of CCl4 back subcutaneous injection and drinking ethanol,the liver fibrosis rat model can be established quickly.The first step of ALPPS operation is safe and feasible in this fibrosis model.The liver of rats with liver fibrosis can still regenerate rapidly after the first step of ALPPS.Background:Previous studies have shown that lipid metabolism plays an important role in liver regeneration.Aquaporin 9(AQP9),as the main glycerol channel in the liver,is mainly involved in liver lipid metabolism,but its mechanism of action in liver regeneration has not been elucidated.In this study,transcriptome sequencing was used to reveal the changes in gene expression during liver regeneration after PVL,and the effect and mechanism of Aqp9 gene knockout on lipid metabolism in liver regeneration were analyzed.Methods:After PVL was done in normal mice and Aqp9 knockout mice,a liver regeneration model was established to observe liver regeneration on the unligated side of mice.Next-generation sequencing was used to reveal the changes in gene expression during liver regeneration after PVL,and the effect of Aqp9 knockout on gene expression of liver regeneration was analyzed.Kyoto encyclopedia of genes and genomes(KEGG),gene oncology(GO)and gene set enrichment analysis(GSEA)were used to further study the changes in signal pathway expression during liver regeneration and the effect of Aqp9 knockout on the liver regeneration pathway.Hematoxylin-eosin staining and Ki67 immunohistochemical staining were performed on liver tissue to observe the morphological changes after regeneration.Results:The liver volume and weight of the unligated side of wild type mice and Aqp9 knockout mice significantly increased after PVL for 3 days.Knockout of Aqp9 had little effect on the increase of liver volume after PVL.The transcriptome was significantly different after PVL,with a total of 1061 genes up-regulated and 296 genes down-regulated.KEGG and GO analysis showed that the expression genes related to cell proliferation and cycle were up-regulated.GSEA analysis showed that the genes related to lipid metabolism were up-regulated after PVL.Knockout of Aqp9 inhibited the up-regulation of some lipid metabolism genes.Both HE staining and Ki67 staining showed that the hepatocytes of the two groups of mice were in a regenerative state 3 days after PVL.Conclusion:This study demonstrated that although knockout of Aqp9 inhibited the upregulation of lipid metabolism-related gene expression during liver regeneration,it did not inhibit liver regeneration after PVL.Background:Previous studies have shown that Aquaporin 9(AQP9)plays an important role in lipid metabolism in nonalcoholic fatty liver disease(NAFLD),but the specific mechanism is unknown.In this study,untargeted metabolomics was used to analyze the role and mechanism of AQP9 in NAFLD.Methods:High-fat diet(HFD)was used to establish NAFLD models in wild type mice and Aqp9 knockout mice.The effects of knockout of Aqp9 on NAFLD were evaluated by measuring body weight,serological indicators,inflammatory factors.Untargeted metabolomics was used to analyze the role and mechanism of AQP9 in NAFLD.Hematoxylin-eosin staining and oil red O staining were used to further verify the morphological changes of the liver after NAFLD modeling and the effect of knockout of Aqp9 on liver fat infiltration.Results:After knockout ofAqp9,the weight gain of mice after HFD was slow,the increase of blood lipid was not obvious,the inflammatory response was lighter.Untargeted metabolomics showed that 220 different metabolites in the liver of mice changed significantly after HFD.Kyoto encyclopedia of genes and genomes(KEGG)analysis showed that glycerophospholipid metabolism and cholesterol metabolism pathways related to the occurrence and development of NAFLD changed significantly.There were 209 different metabolites between Aqp9 knockout mice and wild type mice after HFD.KEGG pathway analysis showed that the knockout of Aqp9 significantly changed the pathway of fatty acid biosynthesis.In the biosynthetic pathway of fatty acid,palmitic acid,stearic acid,capric acid,and myristic acid were significantly reduced,and one lipid precursor acetyl-CoA was increased.Hematoxylin eosin staining and oil red O staining showed that the liver of the two groups of mice showed fat infiltration after feeding HFD,and the liver fat infiltration of Aqp9 knockout mice was lighter.Conclusion:This study demonstrates that knockout of Aqp9 interferes with oxidative stress and lipid metabolism in the development of NAFLD,and can inhibit the development of NAFLD.AQP9 may be a new target for the treatment of NAFLD.