Mechanisms Of Liver Regeneration Regulatory Of ω-3 PUFA Through AMP/LKB1/AMPK Signaling Pathway

Background:Safety limit of the extent of hepatectomy is an effective method for the treatment of large liver tumor. Clinical studies have revealed that precise liver resection technology could increase the resection rate of large liver tumor and tumor at hepatic hilar region. However, weather the patients will survival dependents on restoration and regeneration of the remanent liver. To reveal the mechanism of liver function restoration and liver regeneration, look for more targeted therapeutic targets are of great importance for the safety and effectiveness of complex and difficult liver resection. Postoperative promotion of liver regeneration and restoration of liver function are essential. Large studies reported that ω-3 PUFA protected liver function by suppressing inflammation response. TNF-α、IL-1、IL-6、NO level decreased after liver section when ω-3 PUFA was managed. Recently studies have found that ω-3 PUFA contributed to liver regeneration and enhanced the function of residual liver through regulation of LKB1/AMPK signaling pathway.Objective:Three-dimensional cultured model of primary mouse hepatocytes was established to provide a tool in the research of effects of ω-3 PUFA on bile network formation and to explored the mechanism.Methods:we use the method of three-dimensional culture of primary mouse hepatocytes to observe bile network formation. ω-3 PUFA was used to treat primary hepatocytes and cellular AMP and ATP levels were detected by High Performance Liquid Chromatography at various time-points; Protein levels of LKB1 (liver kinase B1)、p-LKB1、AMPK (AMP-Activated Protein Kinase)、p-AMPK were detected by western blot. Morever, dominant-negative LKB1 and AMPK adenovirus were used to block LKB1 and AMPK pathway respectively and bile network formation was observed.Results:We found that ω-3 PUFA accelerated the formation of bile canaliculi.6 hours after seeding, the hepatocytes attached. After overnight incubation, the hepatocytes extended completely and touched each other. When the hepatocytes were treated withω-3 PUFA DHA, AMP level was increased and ATP level was decreased. AMP/ATP ratio was increased with statistical difference (P<0.05). Moreover, phosphorylation levels of LKB1 and AMPK were elevated (P<0.05). When dominant-negative adenovirus was used to block LKB1 and AMPK pathway, the formation of bile canaliculi were inhibited.Conclusions:In conclusion, ω-3 PUFA promotes bile network formation through AMP/LKB1/AMPK signaling pathway and the result provides theoretical basis for clinical application.