Abstract：
This research explored the pathological and molecular mechanisms of 4-vinylcyclohexene diepoxide (VCD)-induced POI model. QRT-PCR was exploited to detect miR-144 expression in the peripheral blood of POI patients. Rat and KGN cells were treated with VCD to construct POI rat or cell model, respectively. After miR-144 agomir or MK-2206 treatment, miR-144 level, follicle damage, autophagy level and expressions of key pathway-related proteins in rats were detected, and cell viability and autophagy in KGN cells were detected. MiR-144 was apparently down-regulated in the peripheral blood of POI patients. Decreased miR-144 was viewed in both the serum and ovary of rats, yet this trend was apparently reversed by miR-144 agomir. The increased concentration of Follicle-stimulating hormone (FSH) and Luteinizing hormone (LH), along with decreased concentration of E2 and AMH, was observed in the serum of model rats, which was conspicuously negated by control agomir or miR-144 agomir. Increased number of autophagosomes, up-regulated PTEN, and inactivated AKT/m-TOR pathway induced by VCD in ovary tissues were strikingly offset by miR-144 agomir. Results of cytotoxicity assay revealed that 2 mM VCD prominently repressed KGN cell viability. In vitro experiments confirmed that miR-144 interfered with the effect of VCD on autophagy in KGN cells through the AKT/mTOR pathway. Taken together, VCD triggers autophagy to induce POI after targeting the AKT pathway by inhibiting miR-144, it suggest that up-regulation the expression of miR-144 may have the potential to treat POI.
摘要：
本研究探讨了4-乙烯基环己烯二环氧化物（VCD）诱导POI模型的病理和分子机制。采用QRT-PCR检测POI患者外周血miR-144的表达。用VCD处理大鼠和KGN细胞构建POI大鼠或细胞模型,分别。miR-144阿戈米尔或MK-2206治疗后，miR-144水平,卵泡损伤,检测大鼠自噬水平及关键通路相关蛋白的表达,并检测KGN细胞的细胞活力和自噬。在POI患者的外周血中MiR-144明显下调。在大鼠血清和卵巢中观察到miR-144降低,然而这一趋势显然被miR-144agomir逆转.卵泡刺激素（FSH）和黄体生成素（LH）的浓度增加，随着E2和AMH浓度的降低，在模型大鼠的血清中观察到，对照agomir或miR-144agomir明显否定。自噬体数量增加，上调的PTEN,VCD诱导的卵巢组织中失活的AKT/m-TOR通路被miR-144agomir显著抵消。细胞毒性测定的结果表明，2mMVCD显着抑制了KGN细胞的活力。体外实验证实miR-144通过AKT/mTOR通路干扰VCD对KGN细胞自噬的影响。一起来看,VCD通过抑制miR-144靶向AKT通路后引发自噬诱导POI,提示上调miR-144表达可能具有治疗POI的潜力。
