GRIM-19在多囊卵巢综合征颗粒细胞中对VEGF的调控作用及机制研究

Gene associated with retinoid interferon induced mortality 19 (GRIM-19) inhibits signal transducer and activator of transcription-3 (STAT3) mediated signal transduction pathways，which participate in regulating apoptosis and inflammation related pathophysiological process. Vascular endothelial growth factor (VEGF) is closely related to the PCOS by influencing the precess of inflammation and angiogenesis. Our preliminary experiment suggests that GRIM-19 can reduce the expression of VEGF in granulosa cells of PCOS patients, but the mechanism remains to be further explored. Therefore, we propose the hypothesis: GRIM-19 possibly can attenuate VEGF production which cause by hyperandrogenism, or block ROS-HIF1α-VEGF pathway, inhibit the activation of specific transcription factors down-regulating the expression of VEGF, therefore it can prevent or delay the onset of PCOS. To verify this hypothesis, our study will use primary granulosa cells of PCOS patients, PCOS mice model, and techniques such as RT-qPCR, Western blot, and RNA interference. To investigate the molecular mechanisms of how GRIM-19 down-regulate the expression of VEGF. we experiment with the molecular, cellular, tissue and animal field. From the new perspective of GRIM-19, we may lay the foundation for the pathogenesis of PCOS, and further provide new ideas for the prevention and treatment of this disorder.

维甲酸/干扰素诱导细胞凋亡相关基因-19（GRIM-19）通过抑制STAT3介导的信号转导通路参与调节细胞凋亡和炎症相关的病理生理过程。血管内皮生长因子(VEGF)与PCOS密切相关，通过影响炎症反应及血管生成参与PCOS的发生发展。我们预实验提示:GRIM-19能够降低PCOS患者颗粒细胞VEGF表达，但机制尚待进一步探讨。因此我们提出假说：GRIM-19可能通过抑制因高雄激素血症而导致的VEGF增高；或者通过阻断ROS-HIF1α-VEGF通路，抑制特异性转录因子活化，下调VEGF的表达，阻止延缓PCOS发生发展。为验证这一假说，本课题将通过人PCOS原代颗粒细胞、小鼠PCOS模型，采用相关分子生物学技术，在分子、细胞、组织及动物整体水平探讨GRIM-19下调VEGF表达的机制，从GRIM-19新视角为揭示PCOS发生发展机制奠定基础，为治疗PCOS提供新的思路。

维甲酸/干扰素诱导细胞凋亡相关基因-19（GRIM-19）通过抑制STAT3介导的信号转导通路参与调节细胞凋亡和炎症相关的病理生理过程。血管内皮生长因子(VEGF)与PCOS密切相关，通过影响炎症反应及血管生成参与PCOS的发生发展。我们选取PCOS患者、卵巢功能正常患者的卵巢颗粒细胞及人卵巢颗粒细胞癌细胞KGN细胞株作为研究对象，研究PCOS患者卵巢颗粒细胞中GRIM-19对HIF-1α/VEGF通路的影响。结果显示GRIM-19在PCOS卵巢颗粒细胞中表达降低，而HIF-1α、VEGF均表达升高。利用小干扰RNA（Small interfering RNA，siRNA）技术分别下调KGN细胞中GRIM-19 、HIF-1α基因的表达，与阴性对照（Negative control, NC）组对比GRIM-19、HIF-1α、VEGF的信使RNA（mRNA）及蛋白相对表达量的差异。结果显示，在下调GRIM-19组中，GRIM-19的表达量显著降低，HIF-1α、VEGF的表达量显著升高；在下调HIF-1α组中，HIF-1α、VEGF的表达量显著降低，但GRIM-19的表达量与NC对照组差异不明显。表明在KGN细胞中，下调GRIM-19可能通过作用于HIF-1α使VEGF表达增加。同时，我们利用DHEA对小鼠进行PCOS造模，卵巢组织HE染色提示造模成功。与对照组小鼠相比，PCOS小鼠的卵巢组织中，GRIM-19的表达量降低、HIF-1α及VEGF的表达量增加。上述研究结果均表明，卵巢颗粒细胞中GRIM-19可能通过作用于HIF-1α来调控VEGF的表达，GRIM-19作为保护性因子参与PCOS的发生发展，这些研究结果为深入揭示PCOS的发生发展机制，为预防和治疗PCOS提供了新的视角。