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首页> 外文期刊>Reviews of Reproduction >Resistin is a survival factor for porcine ovarian follicular cells
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Resistin is a survival factor for porcine ovarian follicular cells

机译：抵抗素是猪卵巢卵泡细胞的生存因子

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Previously, we demonstrated the expression of resistin in the porcine ovary, the regulation of its expression and its direct effect on ovarian steroidogenesis. The objective of this study was to examine the effect of resistin on cell proliferation and apoptosis in a co-culture model of porcine granulosa and theca cells. First, we analysed the effect of resistin at 1 and 10?ng/ml alone or in combination with FSH- and IGF1 on ovarian cell proliferation with an alamarBlue assay and protein expression of cyclins A and B using western blot. Next, the mRNA and protein expression of selected pro-apoptotic and pro-survival regulators of cell apoptosis, caspase-9, -8 and -3 activity and DNA fragmentation using real time PCR, western blot, fluorescent assay and an ELISA kit, respectively, were analysed after resistin treatment. Furthermore, we determined the effect of resistin on the protein expression of ERK1/2, Stat and Akt kinase. Using specific inhibitors of these kinases, we also checked caspase-3 activity and protein expression. We found that resistin, at both doses, has no effect on cell proliferation. The results showed that resistin decreased pro-apoptotic genes, which was confirmed on protein expression of selected factors. We demonstrate an inhibitory effect of resistin on caspase activity and DNA fragmentation. Finally, resistin stimulated phosphorylation of the ERK1/2, Stat and Akt and kinases inhibitors reversed resistin action on caspase-3 activity and protein expression to control. All of these results showed that resistin has an inhibitory effect on porcine ovarian cell apoptosis by activation of the MAPK/ERK, JAK/Stat and Akt/PI3 kinase signalling pathways.
机译：以前，我们证明了抵抗素在猪卵巢中的表达，其表达的调控及其对卵巢类固醇生成的直接作用。这项研究的目的是在猪颗粒和卵泡膜细胞共培养模型中研究抵抗素对细胞增殖和凋亡的影响。首先，我们使用alamarBlue分析法和western blot分析了单独或与FSH-和IGF1联合使用的抵抗素1和10？ng / ml对卵巢细胞增殖的影响以及细胞周期蛋白A和B的蛋白表达。接下来，分别使用实时PCR，蛋白质印迹，荧光测定和ELISA试剂盒分别选择细胞凋亡，caspase-9，-8和-3活性和DNA片段化的促凋亡和促存活调节剂的mRNA和蛋白表达。抵抗素治疗后进行分析。此外，我们确定了抵抗素对ERK1 / 2，Stat和Akt激酶蛋白表达的影响。使用这些激酶的特异性抑制剂，我们还检查了caspase-3活性和蛋白质表达。我们发现两种剂量的抵抗素都对细胞增殖没有影响。结果表明，抵抗素降低了促凋亡基因，这在所选因子的蛋白表达上得到了证实。我们证明了抵抗素对胱天蛋白酶活性和DNA片段化的抑制作用。最后，抵抗素刺激的ERK1 / 2，Stat和Akt和激酶抑制剂的磷酸化逆转了抵抗素对caspase-3活性和蛋白表达的控制作用。所有这些结果表明，抵抗素通过激活MAPK / ERK，JAK / Stat和Akt / PI3激酶信号传导途径对猪卵巢细胞凋亡具有抑制作用。

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