Dermatologic Unit, Dept. of Health of Sciences, University of Eastern Piedmont, Via Solaroli 17, 28100 Novara, Italy.
Lab. of Physiology, Dept. of Translational Medicine, University of Eastern Piedmont, Via Solaroli 17, 28100 Novara, Italy.
J Dermatol Sci. 2018 Oct;92(1):62-77. doi: 10.1016/j.jdermsci.2018.07.007. Epub 2018 Aug 6.
Estrogens and phytoestrogens can hinder the aging process through mechanisms related to estrogen receptors (ERs), guanine nucleotide-binding protein-coupled receptor (GPER30), mitochondria function and nitric oxide (NO) release. Up to date, however, the above issues are a matter of debate.
To examine the effects elicited by 17 β-estradiol and genistein against peroxidation in human keratinocytes/fibroblasts and evaluate the role played by ERs, GPER30, mitochondria and NO.
Human fibroblasts/keratinocytes, either subjected to peroxidation or not, were exposed to 17 β-estradiol/genistein in the absence or presence of the NO synthase (NOS) inhibitor, the ERs and GPER30 blockers, fulvestrant and G15, the phosphatidyl-inositol-3-kinase (PI3K-Akt), the p38 mitogen-activated protein (MAP) kinase and the extracellular signal-regulated kinases (ERK) 1/2 inhibitors. Specific kits were used for cell viability, NO, ROS and glutathione (GSH) detection and mitochondrial membrane potential measurement. Western Blot analysis was performed for kinases expression/activation detection.
In physiological and peroxidative conditions, 17 β-estradiol/genistein respectively increased and reduced NO release by fibroblasts/keratinocytes. Moreover, both agents prevented the ROS release and the fall of cell viability and mitochondrial membrane potential, while increasing GSH levels and the proliferation rate. Fulvestrant and G15 counteracted all above responses. Also, the NOS, and the kinases blockers reduced the protection exerted by 17 β-estradiol/genistein on cell viability/mitochondria function. The involvement of PI3K-Akt and p38-MAPK was confirmed by Western blot.
17 β-estradiol/genistein protected fibroblasts/keratinocytes against peroxidation by modulating oxidant/antioxidant system and mitochondria membrane potential, through mechanisms related to ERs and GPER30 and kinases activation.
雌激素和植物雌激素可以通过与雌激素受体（ERs）、鸟嘌呤核苷酸结合蛋白偶联受体（GPER30）、线粒体功能和一氧化氮（NO）释放相关的机制来延缓衰老过程。然而，迄今为止，上述问题仍存在争议。
研究 17β-雌二醇和金雀异黄素对人角质形成细胞/成纤维细胞过氧化的影响，并评估 ERs、GPER30、线粒体和 NO 的作用。
将人成纤维细胞/角质形成细胞进行或不进行过氧化处理，然后用 17β-雌二醇/金雀异黄素处理，同时使用一氧化氮合酶（NOS）抑制剂、ERs 和 GPER30 阻滞剂氟维司群和 G15、磷脂酰肌醇-3-激酶（PI3K-Akt）、p38 丝裂原激活蛋白（MAP）激酶和细胞外信号调节激酶（ERK）1/2 抑制剂。使用特定试剂盒检测细胞活力、NO、ROS 和谷胱甘肽（GSH）的释放以及线粒体膜电位。通过 Western Blot 分析检测激酶的表达/激活。
在生理和过氧化条件下，17β-雌二醇/金雀异黄素分别增加和减少了成纤维细胞/角质形成细胞的 NO 释放。此外，这两种药物都能防止 ROS 的释放和细胞活力以及线粒体膜电位的下降，同时增加 GSH 水平和增殖率。氟维司群和 G15 拮抗了所有上述反应。此外，NOS 和激酶抑制剂降低了 17β-雌二醇/金雀异黄素对细胞活力/线粒体功能的保护作用。Western blot 证实了 PI3K-Akt 和 p38-MAPK 的参与。
17β-雌二醇/金雀异黄素通过调节氧化应激/抗氧化系统和线粒体膜电位，保护成纤维细胞/角质形成细胞免受过氧化损伤，其机制与 ERs 和 GPER30 以及激酶的激活有关。
