In Vitro and In Silico Evaluation of Antiurolithiatic Activity of Selected Indigenous Medicinal Plants of Manipur Against Calcium Oxalate Crystallization

Abstract

Urolithiasis, or kidney stone disease, represents a significant global health burden characterized by high recurrence rates and increasing prevalence due to dietary, metabolic, and environmental factors. Calcium oxalate (CaOx) stones account for nearly 80% of all renal calculi. Despite advances in surgical and pharmacological interventions, limitations such as recurrence, high cost, and adverse effects necessitate the exploration of alternative therapeutic strategies. Indigenous medicinal plants of Manipur, a biodiversity-rich region in Northeast India, have long been utilized in traditional healthcare systems for the treatment of urinary disorders, including urolithiasis. However, scientific validation of these ethnomedicinal claims remains insufficient.
The present study investigates the antiurolithiatic potential of selected indigenous medicinal plants of Manipur through an integrated approach combining phytochemical analysis, in vitro crystallization assays, and in silico molecular docking studies. Plant extracts were prepared using aqueous and organic solvents and subjected to qualitative and quantitative phytochemical screening. Antiurolithiatic activity was evaluated using nucleation, crystal growth, and aggregation inhibition assays of calcium oxalate crystals. Advanced analytical techniques, including High-Performance Liquid Chromatography (HPLC) and Gas Chromatography–Mass Spectrometry (GC-MS), were employed to identify bioactive compounds. Molecular docking studies were conducted to assess the interaction of identified phytochemicals with key proteins implicated in stone formation.
Results demonstrated significant inhibition of calcium oxalate crystallization, with methanolic extracts exhibiting the highest activity. Phytochemical analysis revealed the presence of flavonoids, phenolics, and terpenoids, which correlated strongly with inhibitory effects. In silico studies further confirmed strong binding affinities of selected compounds to molecular targets, suggesting potential mechanisms of action. The study validates traditional knowledge systems and highlights the potential of indigenous medicinal plants as sources of novel antiurolithiatic agents.