子宫腺肌症中自噬的调控分子机制及作用的研究

Autophagy is implicated in a variety of gynecological diseases. However, its regulatory mechanism and its role in adenomyosis (ADM) are not fully understood. Our preliminary experiments suggested that autophagy might be activated in the eutopic endometrium of ADM and Hedgehog (Hh) pathway was significantly inhibited. Meanwhile, we also found that Hh pathway inhibition might decrease EGFR phosphorylation in vitro. Previous reported works have suggested that active EGFR-mediated Beclin 1 tyrosine phosphorylation suppresses autophagy activation. Thus, we hypothesized that the suppression of Hh pathway in ADM inhibits EGFR phosphorylation, thereby decreasing tyrosine phosphorylation of Beclin 1 and then cause autophagy activation. In this project, a human primary endometrial stromal cell culture system will be established, and various techniques such as transmission electron microscopy and Duolink in situ PLA will be employed to investigate the molecular mechanism of autophagy regulation. Furthermore, the effect of autophagy on intracellular oxidative stress, and cell apoptosis, migration and invasion will be also studied. Additionally, the role of autophagy in the pathogenesis of ADM will be investigated with a tamoxifen-induced ADM mouse model. The outcome of this project will expand our understanding of Hh-mediated autophagy in ADM. And it will provide us a theoretical basis for the development of potential therapeutic drugs and novel therapeutic strategies for ADM.

自噬在众多妇科疾病的发生与发展中扮演了重要的角色，但其在子宫腺肌症（ADM）中的调控机制及作用尚不明确。我们的预实验发现ADM中自噬可能被激活，且Hedgehog（Hh）通路被显著抑制；在体外实验中我们发现抑制Hh通路可能会干扰EGFR的磷酸化。文献表明活化的EGFR能够通过蛋白相互作用介导Beclin 1酪氨酸磷酸化而抑制自噬。因此，我们推测ADM中Hh通路的抑制会减弱EGFR的磷酸化，从而降低后者对Beclin 1的酪氨酸磷酸化，激活自噬。基于该假说，本课题将利用人原代子宫内膜间质细胞，借助分子生物学、电镜及Duolink® PLA等技术，阐明ADM中自噬的调控机制；同时，我们将研究自噬对子宫内膜细胞氧化应激、凋亡及迁移侵袭能力等的影响，并利用ADM小鼠模型，阐明其在ADM进展中的作用。本课题研究结果有助于加深我们对ADM中自噬的理解，并将为进一步深入探讨ADM的发病机制提供新的思路。

细胞自噬在众多的妇科疾病的发生与发展中扮演了重要的角色，但其在子宫腺肌症（ADM）和子宫内膜异位症（EDM）中的调控机制及作用尚未阐明。本研究围绕这一问题，从组织、细胞与分子层面进行了研究，取得了如下结果：（1）我们发现ADM和EDM的内膜组织中Indian hedgehog（Ihh）信号通路被显著抑制，而它的抑制可显著激活细胞保护性自噬；（2）ADM和EDM异位病灶均存在铁过载现象。我们发现铁过载能显著抑制子宫内膜间质细胞增殖，而在异位病灶处的组织中，我们观察到细胞增殖活力显著降低，提示铁过载在异位病灶细胞增殖活力抑制中起到重要的作用；（3）铁过载能显著激活子宫内膜间质细胞自噬，而自噬的活化能够缓解铁过载诱导的细胞增殖抑制；（4）通过诱导芬顿反应，激活剧烈的细胞内氧化应激，我们发现自噬能通过抑制氧化应激，以减少细胞凋亡。（5）铁过载能够抑制内膜中PARP1的表达，并促进SIRT1的表达，从而调控自噬的活化，并参与对细胞增殖的影响。综上所述，我们的结果表明，子宫内膜细胞自噬能被Ihh信号通路的下调和铁过载所激活，自噬能增强细胞增殖能力，加强细胞抵御氧化应激的能力，从而促进异位病灶处的细胞的存活，促进病灶的持续存在和发展。我们的研究将为更好理解Ihh信号通路及铁过载在内膜细胞自噬中的调控作用及其在ADM和EDM发病机制中的作用提供实验依据，将为利用自噬作为ADM及EDM的治疗靶点提供理论支撑。