Lin Yuanbang, Sheng Mingwei, Weng Yiqi, Xu Rubin, Lu Ning, Du Hongyin, Yu Wenli
The Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road, Tianjin 300052, PR China.
The Department of Anesthesiology, Tianjin First Center Hospital, 24 Fukang Road, Tianjin 300192, PR China.
Biochem Biophys Res Commun. 2017 Feb 5;483(2):885-891. doi: 10.1016/j.bbrc.2017.01.028. Epub 2017 Jan 8.
The effects and mechanism of berberine (BBR) on hepatic injury after orthotopic liver transplantation (OLT) have not been well characterized. We examined the role of Sirt1/FoxO3α axis in the protective effect of BBR on ischemia/reperfusion injury after OLT. Adult male Wistar rats were randomly allocated into four groups: Sham, OLT, OLT with BBR pretreatment (BBR), OLT with BBR and Sirt1 inhibitor (EX527) pretreatment group (EX527). The liver function and oxidative stress level were measured by biochemical and histopathologic examinations. The formation of autophagosome was observed by transmission electron microscopy. The apoptotic rate was determined by TUNEL analysis and the apoptotic mRNA expression. The expression of Sirt1, FoxO3α, Beclin-1, LC3-II/LC3-I, p62 and the acetylation of FoxO3α were assayed by western blot assay and immunoprecipitation. Compared with the OLT group, BBR dramatically attenuated the histopathologic damage, restored the liver function, and decreased the oxidative stress level. Simultaneously, BBR significantly ameliorated apoptosis by decreasing the apoptotic rate and the expression of apoptotic mRNA in rats subjected to OLT. The level of Beclin-1 and LC3-II/LC3-I were upregulated with the inhibition of p62. The deacetylation of FoxO3α by Sirt1 was enhanced in the nuclear of liver after pretreated with BBR. However, the inhibition of Sirt1 by EX527 counteracted the protective effects of BBR. Thus, BBR preconditioning promotes liver transplant ischemia/reperfusion injury partly via activating Sirt1/FoxO3α mediated autophagy.
小檗碱（BBR）对原位肝移植（OLT）后肝损伤的影响及机制尚未完全明确。我们研究了Sirt1/FoxO3α轴在BBR对OLT后缺血/再灌注损伤保护作用中的作用。成年雄性Wistar大鼠随机分为四组：假手术组、OLT组、BBR预处理的OLT组（BBR组）、BBR与Sirt1抑制剂（EX527）预处理的OLT组（EX527组）。通过生化和组织病理学检查测量肝功能和氧化应激水平。通过透射电子显微镜观察自噬体的形成。通过TUNEL分析和凋亡mRNA表达测定凋亡率。通过蛋白质印迹分析和免疫沉淀法检测Sirt1、FoxO3α、Beclin-1、LC3-II/LC3-I、p62的表达以及FoxO3α的乙酰化水平。与OLT组相比，BBR显著减轻了组织病理学损伤，恢复了肝功能，并降低了氧化应激水平。同时，BBR通过降低OLT大鼠的凋亡率和凋亡mRNA表达，显著改善了细胞凋亡。Beclin-1和LC3-II/LC3-I的水平上调，同时p62受到抑制。BBR预处理后，肝脏细胞核中Sirt1介导的FoxO3α去乙酰化增强。然而，EX527对Sirt1的抑制作用抵消了BBR的保护作用。因此，BBR预处理部分通过激活Sirt1/FoxO3α介导的自噬促进肝移植缺血/再灌注损伤。
