Škiljić Dragana, Petersen Anne, Karlsson Jan-Olof, Behndig Anders, Nilsson Staffan, Zetterberg Madeleine
a Department of Clinical Neuroscience/Ophthalmology, Institute of Neuroscience and Physiology , The Sahlgrenska Academy at University of Gothenburg , Gothenburg , Sweden.
b Department of Ophthalmology , Sahlgrenska University Hospital , Mölndal , Sweden.
Curr Eye Res. 2018 May;43(5):639-646. doi: 10.1080/02713683.2018.1437923. Epub 2018 Feb 12.
Protective effects of estradiol against HO-induced oxidative stress have been demonstrated in lens epithelial cells. The purpose of this study was to investigate the effects of 17β-estradiol (E2) on the different superoxide dismutase (SOD) isoenzymes, SOD-1, SOD-2, and SOD-3, as well as estrogen receptors (ERs), ERα and ERβ, in primary cultured human lens epithelial cells (HLECs).
HLECs were exposed to 0.1 µM or 1 µM E2 for 1.5 h and 24 h after which the effects were studied. Protein expression and immunolocalization of SOD-1, SOD-2, ERα, and ERβ were studied with Western blot and immunocytochemistry. Total SOD activity was measured, and gene expression analyses were performed for SOD1, SOD2, and SOD3.
Increased SOD activity was seen after 1.5 h exposure to both 0.1 µM and 1 µM E2. There were no significant changes in protein or gene expression of the different SODs. Immunolabeling of SOD-1 was evident in the cytosol and nucleus; whereas, SOD-2 was localized in the mitochondria. Both ERα and ERβ were immunolocalized to the nucleus, and mitochondrial localization of ERβ was evident by colocalization with MitoTracker. Both ERα and ERβ showed altered protein expression levels after exposure to E2.
The observed increase in SOD activity after exposure to E2 without accompanying increase in gene or protein expression supports a role for E2 in protection against oxidative stress mediated through non-genomic mechanisms.
已在晶状体上皮细胞中证实雌二醇对过氧化氢诱导的氧化应激具有保护作用。本研究的目的是探讨17β-雌二醇（E2）对原代培养的人晶状体上皮细胞（HLECs）中不同超氧化物歧化酶（SOD）同工酶SOD-1、SOD-2和SOD-3以及雌激素受体（ERs）ERα和ERβ的影响。
将HLECs分别暴露于0.1μM或1μM E2中1.5小时和24小时，之后研究其影响。采用蛋白质印迹法和免疫细胞化学法研究SOD-1、SOD-2、ERα和ERβ的蛋白表达及免疫定位。测定总SOD活性，并对SOD1、SOD2和SOD3进行基因表达分析。
暴露于0.1μM和1μM E2 1.5小时后，SOD活性均增加。不同SOD的蛋白或基因表达无显著变化。SOD-1的免疫标记在细胞质和细胞核中明显可见；而SOD-2定位于线粒体。ERα和ERβ均免疫定位于细胞核，并且通过与线粒体追踪染料共定位可见ERβ在线粒体中的定位。暴露于E2后，ERα和ERβ的蛋白表达水平均发生改变。
暴露于E2后观察到SOD活性增加，但基因或蛋白表达未随之增加，这支持E2在通过非基因组机制介导的抗氧化应激中发挥作用。
