Revealing active ingredients, potential targets, and action mechanism of Ermiao fang for treating endometritis based on network pharmacology strategy

• Published in: Journal of ethnopharmacology 2020-10, Vol.260, p.113051-113051, Article 113051
• Main Authors: Zhang, Chao, Su, Tong, Yu, Dan, Wang, Fei, Yue, Chunhua, Wang, Hai-Qiao
• Format: Article
• Language: English
• Subjects: Animals; Anti-Inflammatory Agents - blood; Anti-Inflammatory Agents - pharmacology; Cytokines - blood; Disease Models, Animal; Drugs, Chinese Herbal - pharmacology; Endometriosis - drug therapy; Endometriosis - genetics; Endometriosis - metabolism; Endometriosis - pathology; Endometritis; Ermiao fang; Female; Gene Regulatory Networks; Genomics; Inflammation Mediators - blood; MAPK; Metabolomics; Mitogen-Activated Protein Kinases - metabolism; Network pharmacology; NF-kappa B - metabolism; NF-κB; Protein Interaction Maps; Rats, Sprague-Dawley; Signal Transduction; Signaling pathway; Systems Biology; Uterus - drug effects; Uterus - metabolism; Uterus - pathology
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2020.113051

Ermiao fang (EMF) is a traditional Chinese medicinal herbal formula from ancient times and recorded in the pharmacopeia of the People's Republic of China. It is composed of two typical Chinese herbal medicines, Cortex Phellodendri (Huangbai), the bark of Phellodendron chinensis Schneid. (Rutaceae), and Rhizoma Atractylodis (Cangzhu), the rhizome of Atractylodes lancea (Thunb.) DC. (Compositae). EMF has been clinically used for the treatment of endometritis for many years in China. This study was aimed to identify the active ingredients, potential targets, and mechanism of action of EMF for the treatment of endometritis. In this research, the pharmacological effects of EMF on endometritis were first evaluated by establishing a rat model of endometritis. A network pharmacology-based analytical strategy was then used to predict its targets and signaling pathways. An endometritis-related protein target and compound database was built for EMF. The compounds in EMF and those absorbed into the blood were identified by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). High-throughput virtual screening and molecule docking methods were used to predict the protein targets of EMF. The surface plasmon resonance analysis (SPR) method was used to validate the affinity between the compound and proteins. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was used to predict the related pathways. Western blotting analysis was used to evaluate the expression of key proteins in the related pathways. The animal study showed that EMF could reduce uterine inflammation in rats with endometritis. Then, an ingredient database including 187 compounds and a protein target database including 836 proteins were constructed. Twenty-four compounds in EMF were identified by UHPLC-Q-TOF/MS, among which eight compounds were present in rat plasma after an oral administration of EMF. Afterward, 39 potential target proteins were predicted by the high-throughput screening method, and 20 of them were selected after further screening using molecular docking. Subsequently, an ingredient-target network was constructed, and the target proteins were classified into the NF-κB and MAPK signal pathways by KEGG pathway enrichment analysis. Finally, the affinity between the active ingredients and the target proteins was verified by SPR. The Western blotting analysis showed that EMF significantly inhibited the ele