补肾健脾方介导ER/c-Myc/TERT调控端粒酶活性改善卵巢早衰作用研究

Premature ovarian failure (POF) plays a key role for decreasing telomerase activity, and promoting the expression of TERT gene. It is also regulated by ER/C-Myc/TERT signal pathway, so it is a new way to study antiovarian premature failure drugs. The drugs for Bushen Jianpi recipe were invited by Dr. Xiaguicheng, the master of chinese medicine, show the less side effects compaired with estrogen replacement therapy. We found that these drugs improved the symptoms of POF and raised E2 level. Its mechanism of action related to regulating the hypothalamic-pituitary-gonadal axis ER function. We also found that bcl-2 and p53 were involved in transcription regulation of TERT gene and related to ovarian function. However, the mechanism of the regulation of ER/C-Myc/TERT signal path should be further clarified. In this study, the animal model of ZP3 induced premature ovarian failure and the gene silencing technique in vitro were used to study whether drugs for Bushen Jianpi recipe could promoting the proliferation and development of ovarian granulocytes and the maturation of oocytes through modulationg estrogen and its receptor transduction, stimulating transcription factor C-Myc expression, inducing TERT transcription and increasing telomerase activity. Our study may provides scientific basis for clinical treatment of POF.

卵巢早衰与端粒酶活性相关，TERT是调控端粒酶活性的关键基因，TERT又受ER/c-Myc/TERT信号通路调控，从这一信号通路着手研究药物对它的影响，是抗卵巢早衰药物研究的新方法。补肾健脾方是“国医大师”夏桂成教授防治POF的经验方，与雌激素替代疗法相比副作用明显要小，我们发现它能够显著改善POF患者的症状，提高E2水平，其作用机制与调节下丘脑-垂体-性腺轴的ER功能相关。我们还发现其对bcl-2、P53有明显的调节作用，而bcl-2、P53参与TERT的转录并与卵巢功能有关。但此方对整个ER/c-Myc/TERT信号通路的调控机制有待进一步阐明。本项目用ZP3致卵巢早衰的动物模型及体外基因沉默技术，研究补肾健脾方是否通过调节雌激素及其受体的转导，刺激转录因子c-Myc的表达，诱导TERT转录，增加端粒酶活性，从而促进卵巢颗粒细胞的增殖发育及卵母细胞的成熟，为临床用药提供科学依据。

卵巢功能早衰（POF）是一种常见的妇科疾病，其特征是 40 岁以前出现卵巢功能衰退，伴随着闭经、血清高促性腺激素和低雌激素的症状。患有卵巢功能早衰的女性往往伴随生育力下降或丧失，认知力下降，骨质疏松，心脑血管疾病以及泌尿生殖道症状及性功能减退等症状。卵巢早衰与端粒酶活性相关，TERT是调控端粒酶活性的关键基因，TERT又受ER/c-Myc/TERT信号通路调控，因此，本项目立足这个科学问题以“国医大师”夏桂成教授防治POF的经验方“补肾健脾方”干预自身免疫性卵巢早衰的模型动物以及利用体外基因沉默技术作用于卵巢颗粒细胞，探讨补肾健脾方对ER/c-Myc/TERT信号通路调控的影响，进一步阐明补肾健脾方防治卵巢早衰的作用机制。研究结果：①补肾健脾方可以提高卵巢早衰模型动物以及体外卵巢颗粒细胞的雌激素水平，降低卵巢早衰模型动物的促卵泡刺激素水平；②补肾健脾方提高了卵巢早衰模型动物生长卵泡和成熟卵泡的比例，减少了闭锁卵泡和黄体的数量，改善了卵巢颗粒细胞的水肿；③补肾健脾方可以抑制卵巢颗粒细胞的凋亡；④补肾健脾方明显改善ER/c-Myc/TERT信号通路中关键效应分子ER、c-Myc、TERT蛋白及mRNA的表达，促进卵巢端粒酶活性增加，增加端粒的相对长度。本项目研究结果证实补肾健脾方是通过调节雌激素及其受体的转导，刺激转录因子c-Myc的表达，诱导TERT转录，增加端粒酶活性和端粒相对长度，从而促进卵巢颗粒细胞的增殖发育及卵母细胞的成熟，为扩大名医验方的临床应用，挖掘中药有效物质调节雌激素及其受体的生成提供了科学依据。