# The role and underlying mechanisms of Qi Gong Wan in enhancing the endometrial receptivity of a rat model with polycystic ovary syndrome

## TL;DR

This study explores how Qi Gong Wan improves endometrial receptivity in rats with polycystic ovary syndrome and insulin resistance, potentially through specific genes and compounds.

## Contribution

The study identifies Qi Gong Wan's potential molecular mechanisms and active constituents in improving endometrial receptivity in PCOS-IR rats.

## Key findings

- Qi Gong Wan restores estrous cycles, reduces testosterone and insulin resistance, and improves ovarian morphology in PCOS-IR rats.
- HOXA10, HOXA11, and IGFBP1 are core targets, with wogonin showing strong binding affinity.
- QGW upregulates endometrial receptivity markers and promotes pinopode formation in PCOS rats.

## Abstract

Qi Gong Wan (QGW) is a herbal formula which is used for treating infertility associated with polycystic ovary syndrome (PCOS). However, the mechanism of action remains unclear. This study aimed to investigate whether QGW enhances endometrial receptivity in a PCOS with insulin resistance (PCOS-IR) rat model and to explore the underlying molecular mechanisms and primary active constituents.

A PCOS with insulin resistance (IR) rat model was established using dehydroepiandrosterone (DHEA) and a high-fat diet. Rats were treated with QGW or metformin as a positive control. Network pharmacology and molecular docking were used to identify potential drug-disease targets and active components. Endometrial receptivity was evaluated by assessing key markers—including HOXA10, HOXA11, ITGβ3, LIF, GLUT4, and IGFBP1—using histological examination, scanning electron microscopy (to observe pinopode formation), quantitative real-time PCR, Western blot, and immunohistochemistry. SiRNA-mediated knockdown of Hoxa11 was employed to validate its functional role. Network pharmacology and molecular docking techniques were applied to identify potential drug–disease targets and active constituents.

QGW significantly restored regular estrous cycles, reduced testosterone, fasting insulin, and HOMA-IR levels, and improved ovarian morphology in PCOS-IR rats. Network pharmacological analysis identifies HOXA10, HOXA11, and IGFBP1 as core targets. Molecular docking studies demonstrate that wogonin exhibits strong binding affinity with these targets. QGW upregulates the expression of endometrial receptivity markers (HOXA10, HOXA11, ITGβ3, LIF, GLUT4, IGF1) while downregulating IGFBP1 and IL-6 levels. Additionally, Qigui Wenjing Formula promotes pinopode formation and normalizes estrogen/progesterone receptor expression. When Hoxa11 gene expression is suppressed, this formula can reverse the consequent decline in receptivity-related gene expression.

QGW is capable of enhancing the expression of genes related to endometrial receptivity in PCOS model rats and increasing the number of pinopodes, thereby improving the endometrial receptivity in PCOS rats. The results suggest that QGW may improve endometrial receptivity potentially through upregulation of Hoxa11 accompanied by increased Itgβ3 and decreased Igfbp1. However, whether Hoxa11 directly binds to the promoter regions of relevant genes requires further validation. Network pharmacology and molecular docking suggest that wogonin may be an active constituent with considerable potential, though its specific contribution requires further validation through in vivo and in vitro functional experiments.

The graphical abstract outlines the mechanistic study of Qigong Wan (QGW) for treating polycystic ovary syndrome (PCOS) endometrium.Flowchart detailing the research process on PCOS endometrium. The first panel shows QGW prediction, identifying common targets, hub genes, and enrichment analysis. The second panel presents the effect on endometrial markers in PCOS rats using IHC, ETM, qRT-PCR, and immunoblotting. The third panel involves knocking down endometrial Hoxa11 through qRT-PCR, immunoblotting, and molecular docking. Each step includes diagrams or graphs relevant to the methods used.

The graphical abstract outlines the mechanistic study of Qigong Wan (QGW) for treating polycystic ovary syndrome (PCOS) endometrium.