OBJECTIVE: This study aimed to determine the role of DEX in hepatocyte autophagic flux and liver regeneration after PHx.
METHODS: In mice, DEX was intraperitoneally injected 5 min before and 6 h after PHx. In vitro, DEX was co-incubated with culture medium for 24 h. Autophagic flux was detected by LC3-II and SQSTM1 expression levels in primary mouse hepatocytes and the proportion of red puncta in AML-12 cells transfected with FUGW-PK-hLC3 plasmid. Liver regeneration was assessed by cyclinD1 expression, Edu incorporation, H&E staining, ki67 immunostaining and liver/body ratios. Bafilomycin A1, si-GSK3β and Flag-tagged GSK3β, α2-ADR antagonist, GSK3β inhibitor, AKT inhibitor were used to identify the role of GSK3β in DEX-mediated autophagic flux and hepatocyte proliferation.
RESULTS: Pre- and post-operative DEX treatment promoted liver regeneration after PHx, showing 12 h earlier than in DEX-untreated mice, accompanied by facilitated autophagic flux, which was completely abolished by bafilomycin A1 or α2-ADR antagonist. The suppression of GSK3β activity by SB216763 and si-GSK3β enhanced the effect of DEX on autophagic flux and liver regeneration, which was abolished by AKT inhibitor.
CONCLUSIONS: Pre- and post-operative administration of DEX facilitates autophagic flux, leading to enhanced liver regeneration after partial hepatectomy through suppression of GSK3β activity in an α2-ADR-dependent manner.
目的:本研究旨在确定DEX在PHx后肝细胞自噬通量和肝再生中的作用。
方法:在小鼠中,在PHx之前5分钟和之后6小时腹膜内注射DEX。体外,DEX与培养基共孵育24小时。通过原代小鼠肝细胞中的LC3-II和SQSTM1表达水平以及用FUGW-PK-hLC3质粒转染的AML-12细胞中红色斑点的比例检测自噬通量。通过cyclinD1表达评估肝再生,埃杜公司成立,H&E染色,ki67免疫染色和肝脏/身体比率。巴弗洛霉素A1,si-GSK3β和Flag标记的GSK3β,α2-ADR拮抗剂,GSK3β抑制剂,AKT抑制剂用于鉴定GSK3β在DEX介导的自噬通量和肝细胞增殖中的作用。
结果:术前和术后DEX治疗促进PHx后肝再生,显示比未经DEX治疗的小鼠早12小时,伴随着促进的自噬通量,被巴菲霉素A1或α2-ADR拮抗剂完全废除。SB216763和si-GSK3β抑制GSK3β活性增强DEX对自噬通量和肝再生的影响,被AKT抑制剂废除。
结论:术前和术后给予DEX促进自噬通量,通过以α2-ADR依赖性方式抑制GSK3β活性,导致部分肝切除术后肝再生增强。