PGAM5/Drp1通路促子痫前期滋养细胞坏死样凋亡机制及药物干预研究

Placental trophoblast cells in patients with preeclampsia (PE) have pathological features such as increased necroptosis. Our pilot study found that the expression of phosphoglycerin mutase 5 (PGAM5) was up-regulated and the levels of p-DRP1-Ser637 were down-regulated in PE placenta. The expression of PGAM5 in trophoblast cells was up-regulated under hypoxia, which promoted dephosphorylation of Drp1 at Ser637 and trophoblast cells necroptosis, and these phenomena were reversed by PGAM5 siRNA. Molecular docking found that there is a strong binding between ligustroflavone and PGAM5. Therefore, we raise the point of view for the first time as follow: ligustroflavone reduces trophoblast cells necroptosis in PE through contribution to phosphorylation of Drp1 at Ser637 by inhibiting of PGAM5. We will explore the following contents at clinical, cellular and molecular levels: (1) By using clinical samples, hypoxia-induced trophoblast cells and pregnant rat model of PE with PGAM5 inhibitor and Drp1 inhibitor, we will verify the correlation between PGAM5/Drp1 pathway and PE and clarify the underlying mechanisms for PGAM5/Drp1 pathway in promoting trophoblast cells necroptosis in PE; (2) By using hypoxia-induced trophoblast cells and pregnant rat model of PE, we will clarify the role of ligustroflavonein reducing trophoblast cells necroptosis in PE by inhibiting of PGAM5. This project will contribute to the understanding of the pathogenesis of PE and provide theoretical basis for seeking new targets for drug research and development.

子痫前期（PE）患者胎盘滋养细胞存在坏死样凋亡的病理特征。本项目预实验首次发现PE胎盘中磷酸甘油变位酶5（PGAM5）蛋白水平显著上升、动力相关蛋白1（Drp1）Ser637磷酸化水平显著降低；低氧条件下滋养细胞中PGAM5表达上调，促进了Drp1-Ser637去磷酸化和细胞坏死，沉默PGAM5可取消上述现象。分子对接发现，女贞苷与PGAM5有较强的结合。因此，申请人率先提出女贞苷通过抑制PGAM5，升高Drp1-Ser637磷酸化水平，从而减少PE滋养细胞坏死样凋亡。本项目拟从临床、动物和细胞分子水平重点探讨以下内容：①明确PGAM5/Drp1通路与PE的内在联系，阐明PGAM5/Drp1通路促PE滋养细胞坏死样凋亡机制；②阐明女贞苷通过抑制PGAM5发挥抗PE滋养细胞坏死样凋亡的机制。本项目将有助于阐明PE发生发展的分子机制，为寻找防治PE新靶点提供理论基础。

子痫前期（PE）患者胎盘滋养细胞存在坏死样凋亡的病理特征。我们发现PE胎盘中磷酸甘油变位酶5（PGAM5）蛋白水平显著上升、动力相关蛋白1（Drp1）Ser637磷酸化水平显著降低，并伴随坏死样凋亡相关蛋白的表达升高；为了进一步阐明PGAM5在滋养细胞坏死样凋亡中的作用，我们用PGAM5 siRNA沉默PGAM5。低氧条件下滋养细胞中PGAM5、Drp1表达上调，p-Drp1-S637水平下调；线粒体膜电位和ATP含量降低、活性氧生成增加；细胞坏死增加伴随坏死相关蛋白表达升高。沉默PGAM5可取消上述现象。我们首次证明PGAM5通过Drp1- s637位点去磷酸化激活Drp1，促进缺氧诱导的JEG3细胞坏死，从而导致线粒体功能障碍和坏死相关蛋白表达升高。分子对接分析药物女贞苷与PGAM5有较强的结合，我们在低氧环境中培养人滋养细胞 JEG-3，进行不同浓度女贞苷干预实验，进行各项指标检测，结果发现女贞苷可通过抑制 PGAM5、Drp1表达，缓解线粒体功能障碍，从而延缓低氧诱导的滋养细胞坏死样凋亡。我们的结果为女贞苷作为预防治疗PE新药奠定理论基础和提供新思路。阿司匹林在临床上用于子痫前期的预防和治疗，因此，基于网络药理学和生物信息学分析，突出了阿司匹林预防子痫前期的关键靶点和分子机制。