Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Fertil Steril. 2012 Oct;98(4):1001-8.e1. doi: 10.1016/j.fertnstert.2012.06.008. Epub 2012 Jul 3.
To investigate the protective effect of sphingosine-1-phosphate (S1P) against H(2)O(2)-induced apoptosis in human granulosa cell cultures with freshly harvested granulosa cells.
Experimental study.
Academic medical center for reproductive medicine.
PATIENT(S): Cultures of primary granulosa cells isolated from women undergoing in vitro fertilization (IVF).
INTERVENTION(S): None.
MAIN OUTCOME MEASURE(S): Cell apoptosis and Western blot analysis of signaling pathway proteins.
RESULT(S): We found that S1P (1 and 10 mM) statistically significantly decreased granulosa cell apoptosis after H(2)O(2) treatment. The decreased cell apoptosis induced by S1P was abolished after treatment with VPC23019, an inhibitor of S1P1 and S1P3 receptors, W146, an inhibitor of S1P1 receptors, and CAY10444, an inhibitor of S1P3 receptors. A Western blot analysis revealed that the level of phospho-Akt increased and peaked at 10 minutes after 10 mM S1P exposure.
CONCLUSION(S): Treatment with S1P can inhibit the apoptosis of granulosa cells in response to oxidative stress induced by H(2)O(2). The protective effect of S1P is mediated by activating the PI3K/Akt pathway, and the antiapoptotic effect of S1P is mainly mediated through the S1P1 and S1P3 receptor.
用新鲜采集的人颗粒细胞研究鞘氨醇-1-磷酸（S1P）对人颗粒细胞培养物中 H2O2 诱导凋亡的保护作用。
实验研究。
学术医学中心生殖医学。
接受体外受精（IVF）的女性的原代颗粒细胞培养物。
无。
细胞凋亡和信号通路蛋白的 Western blot 分析。
我们发现 S1P（1 和 10 mM）可显著降低 H2O2 处理后颗粒细胞的凋亡。S1P1 和 S1P3 受体抑制剂 VPC23019、S1P1 受体抑制剂 W146 和 S1P3 受体抑制剂 CAY10444 处理后，S1P 诱导的细胞凋亡减少被消除。Western blot 分析显示，10 mM S1P 暴露 10 分钟后磷酸化 Akt 水平增加并达到峰值。
S1P 处理可抑制 H2O2 诱导的氧化应激对颗粒细胞凋亡的作用。S1P 的保护作用是通过激活 PI3K/Akt 途径介导的，S1P 的抗凋亡作用主要通过 S1P1 和 S1P3 受体介导。
