Surico Daniela, Ercoli Alfredo, Farruggio Serena, Raina Giulia, Filippini Davide, Mary David, Minisini Rosalba, Surico Nicola, Pirisi Mario, Grossini Elena
Gynecologic Unit, Novara, Italy.
Lab. Physiology/Experimental Surgery, Novara, Italy.
Cell Physiol Biochem. 2017;42(3):1051-1062. doi: 10.1159/000478752. Epub 2017 Jun 28.
BACKGROUND/AIMS: estrogens and phytoestrogens exert hepatoprotection through mechanisms not clearly examined yet. Here, we investigated the protective effects exerted by 17β-estradiol and genistein against oxidative stress in hepatocytes and hepatic stellate cells (HSCs) and the involvement of specific receptors and the intracellular signalling.
Huh7.5 and LX-2, alone or in co-culture with Huh7.5, were treated with 17β-estradiol and genistein alone or in the presence of menadione and of estrogen receptors (ERs) and G protein-coupled-estrogenic-receptors (GPER) blockers. Cell viability, mitochondrial membrane potential and oxidant/antioxidant system were measured by specific kits. Western Blot was used for the analysis of Akt and p38-mitogen-activated-protein kinases (MAPK) activation and α-smooth-muscle actin expression.
In Huh7.5, 17β-estradiol and genistein prevented the effects of peroxidation by modulating Akt and p38MAPK activation. Similar antioxidant and protective findings were obtained in LX-2 of co-culture experiments, only. ERs and GPER blockers were able to prevent the effects of 17β-estradiol and genistein.
In Huh7.5 and LX-2, 17β-estradiol and genistein counteract the effects of peroxidation through the involvement of ERs and GPER and by an intracellular signalling related to Akt and p38MAPK. As concerning LX-2, paracrine factors released by Huh7.5 play a key role in protection against oxidative stress.
背景/目的：雌激素和植物雌激素通过尚未明确研究的机制发挥肝脏保护作用。在此，我们研究了17β-雌二醇和染料木黄酮对肝细胞和肝星状细胞（HSCs）氧化应激的保护作用以及特定受体和细胞内信号传导的参与情况。
将Huh7.5细胞和LX-2细胞单独培养或与Huh7.5细胞共培养，分别用17β-雌二醇和染料木黄酮单独处理，或在甲萘醌以及雌激素受体（ERs）和G蛋白偶联雌激素受体（GPER）阻滞剂存在的情况下处理。使用特定试剂盒测量细胞活力、线粒体膜电位和氧化/抗氧化系统。采用蛋白质免疫印迹法分析Akt和p38丝裂原活化蛋白激酶（MAPK）的激活情况以及α-平滑肌肌动蛋白的表达。
在Huh7.5细胞中，17β-雌二醇和染料木黄酮通过调节Akt和p38MAPK的激活来预防过氧化作用。仅在共培养实验的LX-2细胞中获得了类似的抗氧化和保护结果。ERs和GPER阻滞剂能够预防17β-雌二醇和染料木黄酮的作用。
在Huh7.5细胞和LX-2细胞中，17β-雌二醇和染料木黄酮通过ERs和GPER的参与以及与Akt和p38MAPK相关的细胞内信号传导来抵消过氧化作用。关于LX-2细胞，Huh7.5细胞释放的旁分泌因子在抵抗氧化应激中起关键作用。
