Modulation of NFKB ligand RANKL Signaling Lipopolysaccharide Induced Rheumatoid Arthritis Cell Line by Pithecellobium dulce.

Abstract

ntroduction: In the world, 1% of people suffer from chronic inflammatory autoimmune diseases like rheumatoid arthritis (RA). The individual with rheumatoid arthritis suffers from synovial inflammation, cartilage degradation, joint destruction, leading to reduced quality of life. The disease pathogenesis involves the activation of pro- inflammatory cytokines and NF-κB ligand RANKL signaling pathway. The research on plant-based drug design and discovery is to target pathways to find promising phytochemical alternatives for relieving symptoms of RA and other chronic diseases. This study aimed to investigate the role of bioactive compounds Pithecellobium dulce gallic acid and quercetin, in modulating the NF-κB ligand RANKL signaling pathway by targeting MMP-1 expression through in silico and in vitro approaches. Objective: To evaluate the effect of isolated bioactive compounds from Pithecellobium dulce fruit gallic acid, and quercetin on mRNA expression of MMP-1 in NF-kB ligand RANKL signaling. Method: The study involved In Silico and experimental in vitro methods. Ligand– protein interaction studies were conducted to assess gallic acids and quercetin’s therapeutic potential in targeting the NF-κB ligand RANKL signaling pathway. Additionally, green-synthesized Pd-Cu nanoparticles (Pd-CuNPs) were characterized and evaluated. Pharmacokinetic properties, drug-likeness, gastrointestinal absorption, blood-brain barrier permeability, and solubility were predicted using ADME/T tools. Molecular interaction and molecular dynamics simulation were performed to identify potential interactions. The gene expression of MMP-1 was evaluated through RT-PCR in treated rheumatoid arthritis cells. Results: In silico analysis confirmed gallic acid and quercetin have favourable pharmacokinetic and safety profiles. Docking and simulation studies identified strong binding affinities with target proteins. Experimental In vitro results revealed a significant downregulation of MMP-1 mRNA expression in treated RA cells, gallic acid (0.299 ± 0.25, p < 0.05), Pd-CuNPs (0.432 ± 0.22), quercetin (0.519 ± 0.01), and crude extract (0.633 ± 0.03), compared to control. Conclusion: The study demonstrated that gallic acid, quercetin, and Pd-CuNPs exhibit promising modulatory effects on the NF-κB ligand RANKL signaling pathway. These findings support their potential as plant-based alternative agents for managing inflammatory disease like rheumatoid arthritis