Yao Xiaolei, Ei-Samahy M A, Fan Lijie, Zheng Linfeng, Jin Yuyue, Pang Jing, Zhang Guomin, Liu Zifei, Wang Feng
Jiangsu Engineering Technology Research Center of Mutton Sheep and Goat Industry, Nanjing Agricultural University, Nanjing, 210095, PR China.
Jiangsu Engineering Technology Research Center of Mutton Sheep and Goat Industry, Nanjing Agricultural University, Nanjing, 210095, PR China.
Theriogenology. 2018 Jul 1;114:70-80. doi: 10.1016/j.theriogenology.2018.03.014. Epub 2018 Mar 20.
In this study, we investigated the effects of Selenium (Se) on the proliferation of and steroidogenesis in goat luteinized granulosa cells (LGCs) and elucidated the mechanisms underlying these effects. Our results showed that proliferating cell nuclear antigen (PCNA), Akt, and phosphoinositide 3-kinase (PI3K) were expressed mainly in ovarian oocytes and granulosa cells (GCs). We observed that 5 ng/mL Se significantly stimulated LGC proliferation, which could be attributed to increases in PCNA, cyclin-dependent kinase 1 (CDK1), phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK; Thr172), and phosphorylated Akt (p-Akt; Ser473) and decreases in p21 (P < 0.05). Se treatment also significantly increased estradiol (E) production, which could be, at least partially, due to increased levels of 3β-hydroxysteroid dehydrogenase(3β-HSD), steroidogenic acute regulatory protein (StAR), p-Akt (Ser473), and cyclic adenosine monophosphate (cAMP) (P < 0.05); however, follicle-stimulating hormone (FSH) significantly enhanced the production of E, progesterone (P) and cAMP (P < 0.05). Moreover, Se treatment stimulated proliferation and the synthesis of E and cAMP in the presence of FSH (P < 0.05). Additionally, the expression of antioxidant-related genes [glutathione peroxidase (GSH-Px) and superoxide dismutase 2 (SOD2)] and the activity of GSH-Px and SOD were progressively elevated by Se treatment (P < 0.05). These data suggested that Se plays an important role in the proliferation of and steroidogenesis in LGC by activating the PI3K/Akt and AMPK pathways, thereby increasing the expression of its downstream cell-cycle- and steroid-synthesis-related genes, as well as regulating cellular oxidative stress.
在本研究中，我们调查了硒（Se）对山羊黄体化颗粒细胞（LGCs）增殖和类固醇生成的影响，并阐明了这些影响背后的机制。我们的结果表明，增殖细胞核抗原（PCNA）、Akt和磷脂酰肌醇3激酶（PI3K）主要在卵巢卵母细胞和颗粒细胞（GCs）中表达。我们观察到，5 ng/mL的硒显著刺激LGC增殖，这可能归因于PCNA、细胞周期蛋白依赖性激酶1（CDK1）、磷酸化的腺苷酸活化蛋白激酶（p-AMPK；Thr172）和磷酸化的Akt（p-Akt；Ser473）增加以及p21减少（P<0.05）。硒处理还显著增加了雌二醇（E）的产生，这至少部分是由于3β-羟基类固醇脱氢酶（3β-HSD）、类固醇生成急性调节蛋白（StAR）、p-Akt（Ser473）和环磷酸腺苷（cAMP）水平升高（P<0.05）；然而，促卵泡激素（FSH）显著增强了E、孕酮（P）和cAMP的产生（P<0.05）。此外，在FSH存在的情况下，硒处理刺激了增殖以及E和cAMP的合成（P<0.05）。另外，通过硒处理，抗氧化相关基因[谷胱甘肽过氧化物酶（GSH-Px）和超氧化物歧化酶2（SOD2）]的表达以及GSH-Px和SOD的活性逐渐升高（P<0.05）。这些数据表明，硒通过激活PI3K/Akt和AMPK途径在LGC的增殖和类固醇生成中发挥重要作用，从而增加其下游细胞周期和类固醇合成相关基因的表达，并调节细胞氧化应激。
