González Raúl, Ferrín Gustavo, Hidalgo Ana B, Ranchal Isidora, López-Cillero Pedro, Santos-Gónzalez Mónica, López-Lluch Guillermo, Briceño Javier, Gómez Miguel A, Poyato Antonio, Villalba José M, Navas Plácido, de la Mata Manuel, Muntané Jordi
Liver Research Unit, Reina Sofía University Hospital, Córdoba, Spain.
Chem Biol Interact. 2009 Sep 14;181(1):95-106. doi: 10.1016/j.cbi.2009.06.003. Epub 2009 Jun 11.
D-Galactosamine (D-GalN) induces reactive oxygen species (ROS) generation and cell death in cultured hepatocytes. The aim of the study was to evaluate the cytoprotective properties of N-acetylcysteine (NAC), coenzyme Q(10) (Q(10)) and the superoxide dismutase (SOD) mimetic against the mitochondrial dysfunction and cell death in D-GalN-treated hepatocytes. Hepatocytes were isolated from liver resections. NAC (0.5 mM), Q(10) (30 microM) or MnTBAP (Mn(III)tetrakis(4-benzoic acid) porphyrin chloride (1mg/mL) were co-administered with D-GalN (40 mM) in hepatocytes. Cell death, oxidative stress, mitochondrial transmembrane potential (MTP), ATP, mitochondrial oxidized/reduced glutathione (GSH) and Q(10) ratios, electronic transport chain (ETC) activity, and nuclear- and mitochondria-encoded expression of complex I subunits were determined in hepatocytes. d-GalN induced a transient increase of mitochondrial hyperpolarization and oxidative stress, followed by an increase of oxidized/reduced GSH and Q(10) ratios, mitochondrial dysfunction and cell death in hepatocytes. The cytoprotective properties of NAC supplementation were related to a reduction of ROS generation and oxidized/reduced GSH and Q(10) ratios, and a recovery of mitochondrial complexes I+III and II+III activities and cellular ATP content. The co-administration of Q(10) or MnTBAP recovered oxidized/reduced GSH ratio, and reduced ROS generation, ETC dysfunction and cell death induced by D-GalN. The cytoprotective properties of studied antioxidants were related to an increase of the protein expression of nuclear- and mitochondrial-encoded subunits of complex I. In conclusion, the co-administration of NAC, Q(10) and MnTBAP enhanced the expression of complex I subunits, and reduced ROS production, oxidized/reduced GSH ratio, mitochondrial dysfunction and cell death induced by D-GalN in cultured hepatocytes.
D-半乳糖胺（D-GalN）可诱导培养的肝细胞产生活性氧（ROS）并导致细胞死亡。本研究旨在评估N-乙酰半胱氨酸（NAC）、辅酶Q10（Q10）和超氧化物歧化酶（SOD）模拟物对D-GalN处理的肝细胞中线粒体功能障碍和细胞死亡的细胞保护特性。肝细胞从肝切除标本中分离获得。将NAC（0.5 mM）、Q10（30 μM）或锰（III）四（4-苯甲酸）卟啉氯化物（MnTBAP，1mg/mL）与D-GalN（40 mM）共同给予肝细胞。测定肝细胞中的细胞死亡、氧化应激、线粒体跨膜电位（MTP）、ATP、线粒体氧化型/还原型谷胱甘肽（GSH）和Q10比值、电子传递链（ETC）活性以及复合物I亚基的核编码和线粒体编码表达。D-GalN诱导肝细胞线粒体超极化和氧化应激短暂增加，随后氧化型/还原型GSH和Q10比值增加、线粒体功能障碍和细胞死亡。补充NAC的细胞保护特性与ROS生成减少、氧化型/还原型GSH和Q10比值降低以及线粒体复合物I+III和II+III活性及细胞ATP含量的恢复有关。Q10或MnTBAP的共同给药可恢复氧化型/还原型GSH比值，并减少D-GalN诱导的ROS生成、ETC功能障碍和细胞死亡。所研究抗氧化剂的细胞保护特性与复合物I核编码和线粒体编码亚基的蛋白质表达增加有关。总之，NAC、Q10和MnTBAP的共同给药增强了复合物I亚基的表达，并减少了D-GalN在培养肝细胞中诱导的ROS产生、氧化型/还原型GSH比值、线粒体功能障碍和细胞死亡。
