Network Pharmacology and Rat Model Analysis of Yougui Pill in Carrageenan-Induced Prostatitis

Summary

This study integrates network pharmacology and in vivo experiments to investigate the potential effects of Yougui Pill in treating prostatitis. Network pharmacology analysis and YGP treatment in a rat prostatitis model suggested that YGP's therapeutic effects may be associated with the regulation of inflammatory responses, indicating that YGP exerts anti-inflammatory effects in prostatitis.

Abstract

Prostatitis is a common urological disorder with limited treatment options. Yougui Pill (YGP) has shown potential therapeutic value, particularly in inflammation-related conditions. This study aimed to evaluate its effects and explore possible underlying mechanisms. Active components of YGP were screened from TCMSP using oral bioavailability (OB ≥ 30%) and drug-likeness (DL ≥ 0.18) criteria, and corresponding targets were collected. Prostatitis-related targets were obtained from GeneCards, and overlapping targets were analyzed using protein–protein interaction (PPI) networks and Kyoto encyclopedia of genes and genomes (KEGG) enrichment. A total of 126 active compounds and 208 potential targets were identified, with 180 overlapping targets associated with prostatitis. PPI analysis identified TP53, IL6, AKT1, TNF, and IL1B as key hub genes. KEGG enrichment suggested involvement of inflammation-related pathways, including IL-17, TNF, MAPK, and PI3K-Akt signaling pathways. A rat model of prostatitis was established by intraprostatic injection of carrageenan. Histological examination and measurement of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were performed to assess therapeutic effects. YGP treatment improved prostatic histopathology and reduced TNF-α and IL-6 levels at both protein and mRNA levels. These findings suggest that YGP exerts anti-inflammatory effects in prostatitis, while the specific molecular mechanisms remain to be further validated.

Introduction

Prostatitis is a condition where the prostate gland becomes inflamed with swelling and irritation¹. It accounts for approximately 25% of global urological clinical encounters². The National Institutes of Health (NIH) stratified prostatitis into four classifications: acute bacterial, chronic bacterial, chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), and asymptomatic prostatitis³. Among these, CP/CPPS is the most prevalent form and is closely associated with persistent inflammation and immune dysregulation. Despite advances in understanding the inflammatory nature of prostatitis, current....

Protocol

All animal experiments were approved by the Institutional Animal Care and Use Committee of Beijing University of Chinese Medicine (BUCM-2024110107-4103).

Collection of active constituents and targets
YGP consists of Rehmannia glutinosa, Aconitum carmichaelii, Cinnamomum cassia, Dioscorea opposita, Cornus officinalis, Cuscuta chinensis, Angelica sinensis, Eucommia ulmoides, Cervi Cornus Colla, and Lycium barbarum. These herb names were input into the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) t....

Results

Identification of bioactive components and target genes
A total of 126 bioactive components in YGP meeting the criteria (OB ≥ 30% and DL ≥ 0.18) were identified. corresponding to 208 predicted target genes. The GeneCards database yielded 9,845 genes associated with prostatitis. Venn analysis revealed 180 overlapping genes between YGP's predicted targets and prostatitis-related genes, suggesting these shared targets may mediate the therapeutic effects of YGP against prostatitis (F.......

Discussion

Prostatitis is pathologically defined by an increased presence of inflammatory cells within the prostatic parenchyma or mesenchyme^28,29. In a study of over 5 million Korean men followed for nine years, Kim et al. confirmed through multivariate analysis that prostatitis significantly increases the risk of developing prostate cancer³⁰. However, current clinical management remains limited, particularly for non-bacterial prostatitis, where tre.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
4% paraformaldehyde	Beyotime	P0099-3L
AlphaFold Protein Structure Database	AlphaFold Protein Structure Database	https://alphafold.ebi.ac.uk/
BeyoFast SYBR Green qPCR Mix (2X)	Beyotime	D7260
Bioinformatics online platform	Bioinformatics online platform	http://www.bioinformatics.com.cn/
Carrageenan	OriLeaf	S30559
CB-DOCK2	CB-DOCK2	http://183.56.231.194:8001/cb-dock2/php/blinddock.php
Cytoscape	Cytoscape	https://cytoscape.org/
GeneCards	GeneCards	https://www.genecards.org/
Hematoxylin and Eosin (H&E) Staining Kit	Beyotime	C0105S
Metascape	Metascape	https://metascape.org/gp/index.html#/main/step1
Neutral Balsam	Servicebio	WG10004160
Paraffin Wax (for embedding)	Servicebio	RP-15
Protease Inhibitor Cocktail (100X)	Epizyme	GRF101
Rat IL-6 ELISA Kit	Beyotime	PI328
Rat TNF-α Kit	Beijing 4A Biotech Co., Ltd	CRE0003
RNAeasy Animal Long RNA Isolation Kit with Spin Column	Beyotime	R0026
STRING	STRING	https://string-db.org/
TCMSP	TCMSP	https://www.tcmsp-e.com/#/database
Uniprot	Uniprot	https://www.uniprot.org/