Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China.
Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, Hangzhou 310003, Zhejiang Province, China.
World J Gastroenterol. 2017 Oct 14;23(38):6995-7008. doi: 10.3748/wjg.v23.i38.6995.
To investigate the protective mechanism of mitofusin-2 (Mfn2) in rat remote ischemic perconditioning (RIC) models and revalidate it in alpha mouse liver-12 (AML-12) hypoxia cell lines.
Sprague-Dawley rats were divided into three groups ( = 6 each): sham, orthotopic liver transplantation and RIC. After operation, blood samples were collected to test alanine aminotransferase and aspartate aminotransferase. The liver lobes were harvested for histopathological examination, western blotting (WB) and quantitative real-time (qRT)-PCR. AML-12 cell lines were then subjected to normal culture, anoxic incubator tank culture (hypoxia) and anoxic incubator tank culture with Mfn2 knockdown (hypoxia + Si), and data of qRT-PCR, WB, mitochondrial membrane potential (ΔΨm), apoptosis, endoplasmic reticulum Ca concentrations and mitochondrial Ca concentrations were collected.
Both sham and normal culture groups showed no injury during the experiment. The RIC group showed amelioration of liver function compared with the orthotopic liver transplantation group ( < 0.05). qRT-PCR and WB confirmed that Mfn2-mitochondrial Ca uptake 1/2 (MICUs) axis was changed ( < 0.005). In AML-12 cell lines, compared with the hypoxia group, the hypoxia + Si group attenuated the collapse of ΔΨm and apoptosis ( < 0.005). The endoplasmic reticulum Ca decrease and mitochondrial Ca overloading observed in the hypoxia group were also attenuated in the hypoxia + Si group ( < 0.005). Finally, qRT-PCR and WB confirmed the Mfn2-MICUs axis change in all the groups ( < 0.005).
Mfn2 participates in liver injury in rat RIC models and AML-12 hypoxia cell lines by regulating the MICUs pathway.
探讨线粒体融合蛋白 2（Mfn2）在大鼠远程缺血预处理（RIC）模型中的保护机制，并在α肝-12（AML-12）细胞缺氧模型中进行验证。
将 Sprague-Dawley 大鼠分为三组（每组 6 只）：假手术组、原位肝移植组和 RIC 组。手术后采集血样检测丙氨酸转氨酶和天冬氨酸转氨酶。采集肝组织进行组织病理学检查、Western blot（WB）和实时定量（qRT）-PCR。然后将 AML-12 细胞系置于正常培养、缺氧孵箱培养（缺氧）和缺氧孵箱培养加 Mfn2 敲低（缺氧+Si），收集 qRT-PCR、WB、线粒体膜电位（ΔΨm）、凋亡、内质网 Ca 浓度和线粒体 Ca 浓度的数据。
假手术组和正常培养组在实验过程中均无损伤。与原位肝移植组相比，RIC 组肝功能改善（ < 0.05）。qRT-PCR 和 WB 证实 Mfn2-线粒体 Ca 摄取 1/2（MICUs）轴发生改变（ < 0.005）。在 AML-12 细胞系中，与缺氧组相比，缺氧+Si 组 ΔΨm 崩溃和凋亡减轻（ < 0.005）。缺氧组内质网 Ca 减少和线粒体 Ca 超载也在缺氧+Si 组得到缓解（ < 0.005）。最后，qRT-PCR 和 WB 证实所有组的 Mfn2-MICUs 轴均发生改变（ < 0.005）。
Mfn2 通过调节 MICUs 通路参与大鼠 RIC 模型和 AML-12 细胞缺氧模型中的肝损伤。
