| Objective: Liver receives innervation from sympathetic and vagus nervous system.However,whether autonomic innervation changes in hepatic alveolar echinococcosis(AE)or donor liver tissue after liver transplantation have not been reported,and whether autonomic nerves are involved in post-hepatectomy liver regeneration remains unclear.The purposes of this study are as follows:(1)The pathological specimens of AE patients were used to explore the distribution characteristics of autonomic nerve in the lesion,peripheral lesion and normal liver tissue,and its relationship with clinical local stage,liver fibrosis and liver regeneration,to observe the autonomic nerve innervation change in donor liver after liver transplantation.(2)To explore the effects of hepatic vagus nerve repair and regeneration on liver regeneration after hepatectomy in mice and its specific mechanism.(3)To explore the effect and possible mechanism of sympathetic nerve injury on liver regeneration after hepatectomy in rat.Methods: Frist part,(1)The basic clinical information of 74 cases of liver AE was analyzed,and the distribution and characteristics of hepatic sympathetic nerve and vagus nerve in the lesion,adjacent to the lesion and normal liver tissue were detected by pathological specimens.(2)The indexes of pathological changes,fibrosis and cell proliferation in liver lesion,peripheral lesion and normal liver tissue were detected.(3)The correlation between sympathetic nerve and clinical stage,sympathetic nerve and the degree of fibrosis,and the number of cell proliferation was analyzed.(4)Donor liver samples were collected from 2 patients after liver transplantation to detect the characteristics of autonomic nerve regeneration and distribution.Second part,(1)70% hepatectomy and hepatic vagus nerve injury model in mice were established,and liver regeneration,liver index,liver tissue regeneration related cytokines(HGF,VEGF)and liver enzymes(ALT,AST)were detected at different time points after hepatectomy.(2)After supplementation of exogenous netrin-1 protein,as a neuronal growth axon inducer,liver regeneration related indexes and liver enzymes were re-tested.(3)The expression of Ntn1 gene in liver was knocked down by adeno-associated virus(AAV)to detect its effect on liver regeneration.(4)Netrin-1 was added to the mice model of vagus nerve injury and hepatectomy to detect liver regeneration and related indexes of vagus nerve repair and regeneration.(5)Human hepatocyte cell line HL7702 and primary human Schwann cell(HSC)and their co-culture system were induced by netrin-1 to detect the changes of liver cell proliferation.Third part,(1)The 70% hepatectomy and modified hepatic sympathetic nerve injury model by6-hydroxydopamine(6-HODA)in rats was established to detect the indexes of hilar sympathetic nerve injury and liver regeneration.(2)To detect the influence of sympathetic nerve injury model on liver function in rats.(3)To identify the sympathetic nerve in liver tissue and detect the related indexes of nerve regeneration after hepatectomy and sympathetic nerve injury.Results: First part,(1)In 74 patients with hepatic AE,the hepatic sympathetic nerve hyperplasia was obvious in the lesion and adjacent lesion,but there was no change in normal liver tissue,while the vagus nerve did not hyperplasia in the above sites.(2)Sympathetic nerve hyperplasia was active in hepatic AE,which was positively correlated with clinical stage.(3)In terms of pathological changes,multiple rounded larvae were found in the lesions,surrounded by a large amount of cellulose,and a large number of inflammatory cells with active proliferation were observed in peripheral lesions.The degree of sympathetic hyperplasia was positively correlated with the fibrosis of the lesion and peripheral lesions,respectively.And sympathetic hyperplasia was positively correlated with cell proliferation in peripheral lesions.(4)Immunohistochemical staining of PGP9.5,GAP43,TH and VACh T in donor liver after living donor liver transplantation was positive,which was identified as sympathetic nerve by immunofluorescence co-location,while staining of above indexes in remnant liver after auto-transplantation was negative.Second part,(1)hepatic vagal injury significantly inhibited cell proliferation(PCNA,Ki67),liver index,transcription and translation of HGF and VEGF after hepatectomy,and delayed the recovery of ALT and AST.(2)Netrin-1 supplementation promoted liver regeneration after hepatectomy,and promoted the recovery of ALT and AST after hepatectomy.(3)Knockdown of Ntn1 gene significantly inhibited the expression of netrin-1 in liver tissue and delayed liver regeneration after hepatectomy.(4)Netrin-1 supplementation promoted liver regeneration and expression of proteins(NGF,M-ACHR,GAP43,Ch AT,VACh T)in hepatic resection and vagus nerve injury models.Netrin-1 alone did not promote HL7702 cell proliferation,but promoted the proliferation and migration of HSC cells in the co-culture system,and promoted the proliferation of HL7702 cells in co-culture system.Third part,(1)Local hilar ablation with 6-HODA significantly reduced sympathetic nerve distribution,and significantly delayed cell proliferation,liver index and cytokines related to liver regeneration in rats after hepatectomy.(2)6-HODA local sympathetic nerve ablation had no significant effect on liver function.(3)After sympathetic nerve injury,the levels of catecholamine,NGF and netrin-1 in liver tissue were increased in late remnant liver after hepatectomy.Conclusion:(1)In patients with hepatic AE,the hepatic sympathetic nerve showed obvious hyperplasia in and around the lesion,and the degree of hyperplasia was positively correlated with the clinical local stage and the degree of liver fibrosis.Sympathetic hyperplasia around the lesion was positively correlated with the inflammatory cell proliferation.The vagus nerve showed no obvious changes in the lesion,peripheral lesion and normal liver tissue.Sympathetic nerve regeneration and a small amount of vagus nerve regeneration were observed in donor liver after living donor liver transplantation,while no autonomic innervation was detected in remnant liver with hepatic failure after liver auto-transplantation.(2)Hepatic vagus nerve injury significantly inhibits liver regeneration after hepatectomy,and nerve axon-inducer netrin-1 can promote liver regeneration after hepatectomy,possibly by promoting the proliferation and migration of Schwann cells to promote vagus nerve repair and regeneration.(3)Sympathetic nerve injury model can be established by local ablation of hepatic sympathetic nerve,which is a safe and feasible modified sympathetic nerve injury model.Sympathetic nerve injury can significantly delay liver regeneration after hepatectomy and regeneration after sympathetic nerve injury may be involved in liver regeneration after hepatectomy. |
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