基于PI3K/AKT信号通路探讨二仙汤治疗化疗损伤性卵巢早衰的作用机理

Er-xian Decoction is used to treat the premature ovarian failure clinically. In preliminary experiments, we found that Er-xian Decoction could promote the development and maturation of immature rats follicles. On the base of this work,we will explore the mechanism of Er-xian on the primordial follicles onset,the reserve of primordial follicles and apoptosis of granulosa cell from the PI3K/AKT signaling pathway in vivo and in vitro.In vivo,the ovarian pathomorphology will be observed by establishing the animal model of chemotherapy damaged POF. estradiol (E2) and progesterone (P),follicle stimulating hormone (FSH),luteinizing hormone (LH), inhibin B (INH-B) and anti-Mullerian hormone (AMH) will be analysised.We will investigate the function of PTEN gene in the PI3K/AKT pathway.In vitro,primary culture of granulosa cell from the ovaries of rats is provided the cellular research method to further research of POF.Er-xian decodtion serum will effect on the expression of the PI3K/AKT and its phosphorylation and the proliferation and apoptosis of granular cells.The project will clarify that the compound may inhibit the initiation of primordial follicle,increase the reserve of primordial follicle, and inhibit of apoptosis of granulosa cells to improve ovarian function. This study will greatly open up new prospects for the traditional Chinese medicine on the intervention of human POF pathological process.

二仙汤临床上治疗卵巢早衰（POF）疗效确切。在前期实验中我们发现二仙汤能够促进幼年大鼠卵泡发育与成熟。本项目在此基础上通过体内、外实验，从PI3K/AKT信号通路深入研究二仙汤对原始卵泡的储备、启动及对颗粒细胞凋亡的干预机制。用临床疗效确切的二仙汤灌服化疗损伤后POF大鼠模型，观察药物对卵巢病理形态学的影响，分析内分泌指标：雌二醇（E2）、孕激素（P）、卵泡刺激素（FSH）、黄体生成素（LH）、抑制素B（INH-B）和抗苗勒氏管激素（AMH）的变化，及抑癌基因PTEN通过PI3K/AKT通路对原始卵泡储备、启动的调控。体外培养卵巢颗粒细胞，分析药物血清对PI3K/AKT及其磷酸化产物的表达及颗粒细胞增殖与凋亡的变化。阐明该复方可能是通过抑制原始卵泡的启动，增加原始卵泡的储备，抑制颗粒细胞的凋亡，从而达到改善卵巢功能的作用。本研究还将极大地开辟了中医药对人类POF病理过程干预的新前景。

本研究项目通过体内、外实验探索二仙汤治疗化疗性卵巢早衰的作用机制。建立化疗损伤卵巢早衰动物模型，观察二仙汤对卵泡储备和卵巢功能的影响。发现二仙汤能明显改善血清激素水平，增加卵泡膜血管密度，促进颗粒细胞分泌AMH和INH，恢复卵泡正常募集和成熟，减少卵泡闭锁。并且通过抑制卵泡颗粒细胞凋亡改善卵巢储备功能。进一步应用二仙汤含药血清对顺铂干预大鼠卵巢颗粒细胞的影响，明确二仙汤调控凋亡的作用机制。发现二仙汤药物血清能现在改善颗粒细胞活性，降低顺铂导致颗粒细胞凋亡，激活PI3K/AKT信号通路，并使下游作用因子 Bcl-2 上调、 Bax 下调。因此，二仙汤可通过PI3K/AKT信号通路治疗化疗损伤性POF。该工作首次从卵泡储备与颗粒细胞凋亡角度系统研究二仙汤治疗POF的作用机制，为临床科学用药提供理论依据。