Liang Yan, Wang Huimin, Chen Jin, Chen Lingyan, Chen Xiaoyong
Department of Traditional Chinese Medicine, Maternal and Child Health Hospital of Jiangxi Province, Nanchang, Jiangxi 330006, China.
Korean J Physiol Pharmacol. 2023 Mar 1;27(2):167-176. doi: 10.4196/kjpp.2023.27.2.167.
This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.
本研究旨在探讨地黄苷D（RD）对卵巢储备功能减退（DOR）大鼠卵巢功能的影响及其潜在作用机制。单次注射环磷酰胺建立DOR大鼠模型，注射后14天，大鼠灌胃给予RD两周。使用阴道涂片检测大鼠发情周期。对卵巢组织进行组织学评估，计算原始卵泡、成熟卵泡和闭锁卵泡的数量，以及颗粒细胞的凋亡率。通过酶联免疫吸附测定法测定促卵泡生成素（FSH）、促黄体生成素（LH）和雌二醇（E）水平。研究DOR大鼠卵巢组织中叉头框蛋白O1（FOXO1）、Klotho蛋白、Bcl-2和Bax的蛋白水平。通过染色质免疫沉淀法验证FOXO1与Klotho之间的结合。对DOR大鼠高剂量给予RD可改善其发情周期，增加卵巢指数，增加原始卵泡和成熟卵泡数量，减少闭锁卵泡数量，抑制卵巢颗粒细胞凋亡，此外还可抑制FSH和LH水平并上调E的表达。DOR大鼠中FOXO1和Klotho被显著抑制。敲低FOXO1可部分抑制RD对DOR大鼠的保护作用，尽管敲低了FOXO1，但过表达Klotho可恢复RD诱导的对DOR发展的阻断作用。FOXO1抗体富集Klotho启动子，与假手术组相比，DOR组中它们之间的结合减少。RD通过FOXO1/Klotho轴改善DOR大鼠的卵巢功能并减少颗粒细胞凋亡。
