Rhazya stricta Targets Proliferation/Apoptosis/ Inflammation Axis in Osteoarthritis: A Comprehensive Network Pharmacology Based Experimental Validation Study

Background

Osteoarthritis (OA), a degenerative joint disorder, demands multi-target therapeutic strategies. Rhazya stricta, a traditional medicinal plant, was explored for its phytochemicals and their anti-OA potential.

Objectives

To investigate Rhazya stricta's anti-OA mechanisms via integrated network pharmacology and experimental validation.

Materials and Methods

Phytochemicals from IMPPAT and KNAPSACK databases (39 compounds) underwent ADME/toxicity screening (SwissADME, Molsoft, ProTox-II), yielding 10 non-toxic candidates. SwissTargetPrediction and GeneCards identified OA-associated targets, intersected via Venny 2.0. STRING analyzed Protein-Protein Interactions (PPI), while Cytohubba prioritized hub genes. Functional enrichment (ShinyGo) mapped pathways, and Cytoscape visualized compound-targetpathway networks. Experimental validation included MTT assay (cytotoxicity) and qRT-PCR (gene expression) in SW-982 cells.

Results

Phytochemical profiling identified 10 bioactive compounds with favorable pharmacokinetics. Intersection of 286-compound targets and 770-OA genes revealed 86 shared targets. PPI network analysis prioritized CASP3, ESR1, EGFR, PPARG, and MMP9 as hub genes, linked to PI3K-Akt, MAPK, and inflammation pathways. KEGG enrichment highlighted OA-relevant pathways (e.g., calcium signaling, EGFR resistance). Cytoscape networks demonstrated multi-target interactions. Experimentally, Rhazya stricta extract exhibited dose-dependent cytotoxicity (IC₅₀: 45 µg/mL) and modulated hub gene expression: CASP3 upregulated (2.6-fold, p=0.001), while ESR1 and EGFR downregulated (0.41-and 0.34-fold, p=0.001), aligning with bioinformatics predictions. RABG2 dataset analysis confirmed elevated CASP3, ESR1, and EGFR in OA tissues, with significant gene correlations (ESR1-CASP3: r=0.38; CASP3-EGFR: r=-0.12; p<0.001).

Conclusion

Rhazya stricta targets OA through synergistic modulation of apoptosis (CASP3), inflammation (ESR1), and proliferation (EGFR) via key pathways, validated experimentally. This study underscores its potential as a multi-target therapeutic agent for OA, bridging computational predictions with biological evidence.