Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

Yan Liang; Huimin Wang; Jin Chen; Lingyan Chen; Xiaoyong Chen

doi:10.4196/kjpp.2023.27.2.167

Indexed in SCIE, Scopus, PubMed & PMC

pISSN 1226-4512 eISSN 2093-3827

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Korean J Physiol Pharmacol 2023; 27(2): 167-176

Published online March 1, 2023 https://doi.org/10.4196/kjpp.2023.27.2.167

Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

Yan Liang, Huimin Wang, Jin Chen, Lingyan Chen, and Xiaoyong Chen*

Department of Traditional Chinese Medicine, Maternal and Child Health Hospital of Jiangxi Province, Nanchang, Jiangxi 330006, China

Correspondence to:Xiaoyong Chen
E-mail: chenxiaoyong668899@163.com

Author contributions: Y.L. and X.C. conceived the ideas. Y.L., X.C., and H.W. designed the experiments. Y.L., X.C., H.W., and J.C. performed the experiments. Y.L., X.C., and H.W. analyzed the data. Y.L., X.C., H.W., J.C., and L.C. provided critical materials. Y.L., H.W., and J.C. wrote the manuscript. Y.L. and X.C. supervised the study. All the authors have read and approved the final version for publication.

Received: July 11, 2022; Revised: August 22, 2022; Accepted: September 19, 2022

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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Abstract

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 (E₂) 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 E₂ 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.

Keywords: Forkhead Box O1, KLOTHO, Ovarian reserve, Rehmannioside D