Role of Clitoria ternatea (Butterfly Pea) Flower in Endometriosis and Related Pain: A Network Pharmacology-Based Investigation and Experimental Validation ^†

Abstract

This study explored the potential role of Clitoria ternatea (CT) flower in ameliorating endometrial pain (EP) through network pharmacology and experimental approaches. Phytochemicals of the CT flower were listed from the literature and databases, and 18 suitable actives were screened for bioavailability and drug likeness parameters using SwissADME. For these actives, 279 exclusive target genes were predicted using SwissTargetPrediction. Additionally, 939 exclusive genes for EP were acquired from the DisGenet and GeneCards databases. Ninety-one overlapping gene targets of CT and EP were listed, for which a Protein–Protein Interaction (PPI) network was constructed using STRING. The top three node proteins (SRC, ESR1, and PI3KR1) in the PPI network were identified through Cytoscape (version 3.9.1). Molecular docking analysis of the eighteen actives with the three target proteins showed strong binding interactions of Flavylium, kaempherol, and quercetin with all the targets, suggesting their involvement in EP relief. In addition, Gene Ontology (GO) functions analysis revealed 320 biological processes, 59 cellular components, and 107 molecular functions were enriched with the target genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses identified 106 KEGG pathways, including steroid hormone biosynthesis, endocrine resistance, and endometrial cancer pathways, which were significantly enriched with the target genes. The anti-inflammatory and analgesic effects of CT’s methanolic extract (ME) were investigated through in vitro and in vivo assays. The ME exhibited 91.47% inhibition of heat-induced hemolysis compared to 92.87% by aspirin in the in vitro membrane stabilizing assay. The in vivo carrageenan-induced paw edema study revealed 65.28% inhibition of paw edema by ME compared to 80.38% inhibition by aceclofenac at the end of 4-h treatment. The in vivo acetic acid-induced writhing test demonstrated analgesia by ME by 75.6% inhibition of writhing compared to 77.49% by aceclofenac. These findings suggest CT flower could be a potential natural remedy for EP, warranting further investigation in future studies.