红参对卵巢储备功能减退大鼠卵巢储备功能和卵巢组织磷脂酰肌醇 3-激酶/蛋白激酶 B/哺乳动物雷帕霉素靶蛋白信号通路的影响

目的探讨红参对卵巢储备功能减退(DOR)大鼠卵巢功能的改善作用以及潜在机制。方法 2022年 8—11月进行 DOR大鼠模型建立实验。将 48只 SD雌性大鼠分为空白组、模型组、模型+雌二醇组以及模型+红参组，每组 12只。采用腹腔注射去氧乙烯基环己烯(VCD)225 mg/kg建立 DOR大鼠模型。模型组与空白组均给予 0.9%氯化钠溶液灌胃，模型组+雌二醇组给予 0.15 mg/kg戊酸雌二醇溶液灌胃，模型+红参组给予 5 g/kg红参混悬液灌胃，1次/天，共 30 d。 30 d后比较各组大鼠治疗前后体质量变化；观察卵巢组织病理切片变化；检测血清促卵泡生成素(FSH)、促黄体生成素(LH)、雌二醇、促性腺激素释放激素(GnRH)和抗缪勒管激素(AMH)激素水平变化；蛋白质印迹法检测卵巢组织中磷脂酰肌醇 3-激酶 p85α(PI3K p85α)、蛋白激酶 B(Akt)和哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达量和磷酸化水平。结果用药后，模型+雌二醇组大鼠体质量差值为(15.0±0.7)g高于模型组的(10.9±1.3)g，体质量明显增加(P<0.05)。模型+红参组大鼠体质量差值为(14.9±0.4)g，高于模型组的(10.9±1.3)g(P<0.05)，但与模型+雌二醇组的(15.0±0.7)g比较差异无统计学意义(P>0.05)。病理切片显示，模型+雌二醇组和模型+红参组卵泡数量多于模型组，卵巢结构较为清晰，颗粒结构更为紧凑，黄体数量较模型组增多；ELISA实验显示，用药后，模型+红参组和模型+雌二醇组 AMH、雌二醇和 GnRH明显升高，FSH、LH明显降低(P<0.05)其中模型+红参组激素水平恢复更明显(P<0.05)。蛋白质印迹法显示，与模型组比较，用药后，模型+红参组和模型+雌二醇组，p-PI3K p85α、p-Akt和 pmTOR均显著下降(P<0.05)；与雌二醇组比较，模型+红参组 p-PI3K p85α、p-Akt、p-mTOR水平降低(P<0.05)。结论红参可以有效改善 DOR模型大鼠的内分泌，可能通过调控 PI3K/Akt/mTOR信号通路参与改善卵巢功能。

Effect of red ginseng on improving ovarian reserve function and PI3K / Akt / mTOR signaling pathway in ovarian tissue of rats with decreased ovarian reserve

Objective To explore the effect of red ginseng on improving ovarian function in rats with decreased ovarian reserve (DOR).Methods DOR rat model building experiments were performed between August 2022 and November 2022. Forty-eight SD female rats were randomly assigned into blank group, model group, model + estradiol group, and model + red ginseng group with 12 micein each group. The DOR rat model was established by an intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD) 225 mg/kg.Then both model group and blank group were given normal saline gavage, model group + estradiol group received 0.15 mg/kg valeric acid solution, and model + red ginseng group received 5 g/kg red ginseng suspension, once a day, 30 days in total. After 30 days, changesin body mass before and after treatment were compared, changes in ovarian histopathological sections were observed, and serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, gonadotropin releasing hormone (GnRH) and anti-mullerian hormone (AMH) levels were tested. Western blotting detection was performed of phosphatidylinositol 3-kinase p85α (PI3K p85α), protein kinase B (Akt) and mammalian target of rapamycin (mTOR) in ovarian tissue.Results After medication, the difference in body mass in the model + estradiol group was (15.0±0.7) g, which was higher than that [(10.9±1.3) g] in the model group (P <0.05). The difference inbody mass in the model + red ginseng group was (14.9± 0.4) g, which was significantly higher than that [(10.9±1.3) g] in the modelgroup (P <0.05), but statistics were not significant in comparison with the difference in model+estradiol group [(15.0±0.7) g] (P>0.05).The pathological section showed that the number of follicles in the model + estradiol and model + red ginseng groups was more thanthat in the model group, with clear ovarian structure and more compact granular structure, and the number of corpus luteum more thanthat in the model group. ELISA experiment showed that AMH, estradiol and GnRH levels in the model + red ginseng group and model+ estradiol group significantly were increased, while FSH and LH levels were significantly decreased (P<0.05), and the hormone level in the model + red ginseng group recovered more significantly (P<0.05). Western blotting results showed that p-PI3K p85α, p-Akt and p-mTOR levels were significantly decreased in model + red ginseng group and model + estradiol group in comparison with the modelgroup (P<0.05), and p-PI3K p85α, p-Akt and p-mTOR levels were decreased in model + ginseng group in comparison with model + estradiol group (P<0.05).Conclusion Red ginseng can effectively improve the endocrine of DOR model rats, and may participate in improving ovarian function by regulating PI3K / Akt / mTOR signaling pathway.