P-686 Molecular mechanism of Zishen Yutai Pills protecting ovarian reserve in premature ovarian insufficiency: a multi-omics analysis
Authors: Feng Y , Chen Y , Zhong Z , Zhao Y 
Published: June 2022

Abstract

Study question
What is the potential molecular mechanism of Zishen Yutai Pills when compared to Hormone Replacement Therapies in treating premature ovarian insufficiency through protecting ovarian reserve?

Summary answer
Compared to estrogen, the cAMP signaling pathway and steroid hormone biosynthesis may be the potential molecular mechanisms for Zishen Yutai Pills preserving ovarian function.

What is known already
Premature ovarian insufficiency (POI) is a major reproductive disease that greatly affects ovarian function. It is a complex disorder with multiple pathogenic factors and high etiological heterogeneity. The conventional treatment for POI is hormone replacement therapies (HRT), which could alleviate the low estrogen syndromes. Compared to HRT, Zishen Yutai Pills (ZSYTP), a patented formula that is widely used for treating POI, can induce follicle development and stimulate ovulation. The combination of omics, including proteomics, metabolomics and transcriptomics, could promote the comprehensive understanding of the potential molecular mechanism involved in both effective treatments for POI in the rat model.

Study design, size, duration
20 female Sprague-Dawley rats were randomized to 4 groups: the normal control group (NC), POI model group (POI), ZSYTP group and E2 Gavage group(E2). Intraperitoneally injected with cyclophosphamide for 15 days to establish a POI rat model, follow up with intragastric administration with ZSYTP or E2 daily for 3 weeks. The combined analysis of the proteomics, metabolomics and transcriptomics of rat ovarian tissues shows the potential pathways and molecular targets involved in ZSYTP and E2.

Participants/materials, setting, methods
All protocols of animal experiments were approved by the Institutional Animal Care and Ethics Committee of Zhuhai BestTest (IAC (S) 202108005). Rat ovary tissues were weighed and quickly frozen after dissection. Muti-omics analysis was performed based on the results from proteomics, metabolomics and transcriptomics analysis, in which Correlation and KEGG pathway analysis were conducted. Statistical analyses were performed with SPSS; statistical significance was set to P &lt; 0.05.

Main results and the role of chance
Both the E2 and ZSYTP groups show elevated serum estrogen levels compared with the POI group(P&lt;0.05). In the histological assessment, a significantly elevated amount of primary follicles was observed in the ZSYTP group compared to the E2 POI group, indicating that ZSYTP improves ovarian follicle development. Under electron microscopy, fewer necrosis cells were observed in the ZSYTP group, which shows that ZSYTP could improve ovarian repair function and reserve. A combined analysis of ovary proteomics, metabolomics and transcriptomics in ZSYTP vs. E2 shows that both interventions share molecular functions, including steroid binding, protein digestion and absorption, and carbon metabolism. In the KEGG pathway enrichment analysis, the cAMP signaling pathway and steroid hormone biosynthesis were enriched.

Limitations, reasons for caution
A larger sample size of animal experiments is needed for future study. Quantitative expression analyses of specific genes disclosed in this multi-omics analysis are the next stage of the study.

Wider implications of the findings
This is the first study to conjoint analyses multi-omics to explore the two different treatments on POI. Zishen Yutai Pills could alleviate low estrogen syndrome and preserve ovarian function through regulating the cAMP signaling pathway and steroid hormone biosynthesis. Zishen Yutai Pills could serve as an effective treatment for POI.

Trial registration number
Not applicable