β-Sitosterol-loaded leciplexes attenuate rheumatoid arthritis in rats by modulating JAK2/STAT3, NF-κB, and p38 MAPK...

Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by persistent synovial hyperplasia, progressive osteoarticular destruction, and systemic inflammatory complications. β-Sitosterol (BSS), a plant-derived phytosterol, exhibits anti-inflammatory and antioxidant properties; however, its therapeutic utility is limited by poor aqueous solubility and low oral bioavailability. This study aimed to enhance the oral delivery of BSS using Leciplex nanocarriers (BSS-LPXs) and to investigate their effects on key inflammatory pathways in Complete Freund's Adjuvant (CFA)-induced RA in rats. BSS-LPXs were optimized using a Box-Behnken design to achieve optimal vesicle size, entrapment efficiency, and sustained drug release. Pharmacokinetic parameters were assessed following oral administration in rats. Anti-arthritic pharmacodynamics were evaluated via clinical arthritis scoring, serum cytokine levels (TNF-α, IL-6), oxidative stress markers (GSH, SOD, MDA), and histopathological examination. Mechanistic studies focused on the modulation of JAK2/STAT3, NF-κB, and p38 MAPK signaling pathways. Optimized BSS-LPXs had an average size of ~ 146 nm, 66% entrapment efficiency, and sustained release characteristics. Oral administration of BSS-LPXs significantly increased bioavailability by 3.8-fold compared to the conventional suspension (p < 0.05), with higher C_max and prolonged half-life. In vivo, BSS-LPXs attenuated arthritis progression, preserved body weight, lowered TNF-α and IL-6 levels, restored antioxidant defenses, and decreased lipid peroxidation. Mechanistically, the formulation downregulated JAK2/STAT3 expression and suppressed the activation of NF-κB and p38 MAPK signaling. Histopathological analysis confirmed reduced synovial hyperplasia and cartilage damage. Leciplexes improved oral bioavailability and systemic exposure of BSS compared with conventional suspension, while enhancing anti-arthritic efficacy by modulating key inflammatory pathways, reducing oxidative stress, and protecting joint structure. This combined pharmacokinetic and pharmacodynamic profile highlights the potential of LPXs as an effective strategy to improve the delivery and anti-arthritic activity of hydrophobic phytoconstituents.