Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Nitra, Slovak Republic.
J Trace Elem Med Biol. 2011 Jul;25(3):166-70. doi: 10.1016/j.jtemb.2011.05.002. Epub 2011 Jun 23.
The general objective of this in vitro study was to examine the secretory activity (insulin-like growth factor I, IGF-I) of porcine ovarian granulosa cells after Ag addition and to outline the potential intracellular mediators (cyclin B1 and caspase-3) of its effects. Ovarian granulosa cells were incubated with silver nitrate (AgNO(3)) at the doses 0.09, 0.17, 0.33, 0.5 and 1.0 mg/mL for 18 h and compared to the control group without metal addition. The release of IGF-I by granulosa cells was assessed by RIA and expression of cyclin B1 and caspase-3 immunocytochemistry. Our observations show that IGF-I release by granulosa cells was significantly (P<0.05) stimulated by AgNO(3) addition at the doses (0.09-1.0 mg/mL). Similarly to IGF-I the cyclin B1 and caspase-3 expression in ovarian granulosa cells was stimulated by Ag addition (0.09-1.0 mg/mL). In conclusion, the present results indicate, a direct effect of Ag on (1) secretion of growth factor IGF-I, (2) expression of markers of proliferation (cyclin B1) and apoptosis (caspase-3) of porcine ovarian granulosa cells and (3) that the effect of Ag on ovarian cell proliferation could be mediated by IGF-I and cyclin B1. Obtained data indicate the interference of Ag in the pathways of proliferation and apoptosis of porcine ovarian granulosa cells through hormonal and intracellular peptides such as are cyclin B1 and caspase-3.
本体外研究的总体目标是检测添加银后猪卵巢颗粒细胞的分泌活性（胰岛素样生长因子 I，IGF-I），并概述其作用的潜在细胞内介质（细胞周期蛋白 B1 和半胱天冬酶-3）。将卵巢颗粒细胞与硝酸银（AgNO3）在 0.09、0.17、0.33、0.5 和 1.0 mg/mL 的剂量下孵育 18 小时，并与未添加金属的对照组进行比较。通过 RIA 评估颗粒细胞释放的 IGF-I，并通过免疫细胞化学检测细胞周期蛋白 B1 和半胱天冬酶-3 的表达。我们的观察结果表明，AgNO3 的添加以剂量（0.09-1.0 mg/mL）显著（P<0.05）刺激颗粒细胞释放 IGF-I。与 IGF-I 类似，Ag 的添加（0.09-1.0 mg/mL）也刺激卵巢颗粒细胞中细胞周期蛋白 B1 和半胱天冬酶-3 的表达。总之，目前的结果表明，Ag 对（1）生长因子 IGF-I 的分泌、（2）猪卵巢颗粒细胞增殖标志物（细胞周期蛋白 B1）和凋亡（半胱天冬酶-3）的表达有直接影响，（3）Ag 对卵巢细胞增殖的影响可能通过 IGF-I 和细胞周期蛋白 B1 介导。获得的数据表明，Ag 通过激素和细胞内肽（如细胞周期蛋白 B1 和半胱天冬酶-3）干扰猪卵巢颗粒细胞的增殖和凋亡途径。
