Assessment of Wound Healing Potential of Punica granatum Peel Extract in Rheumatoid Arthritis Mice: In silico and In Vivo Study

Rheumatoid arthritis (RA) is the most widespread chronic autoimmune disorder worldwide, which is known to hinder wound healing. Therefore, this study aimed to investigate the effectiveness of pomegranate peel extract (PPEx) in accelerating wound healing in rheumatoid arthritis mice model through both in silico and in vivo approaches. Gas chromatography-mass spectrometry (GC-MS) analysis identified 31 active compounds in PPEx. The in-silico analysis explored the interaction between PPEx active compounds and inflammatory proteins (TNF-α, IL-6, NF-κB p65, RORγt, and T-bet) using the molecular docking method. Seven active compounds out of 31 had higher binding affinity, four of them (Lovastatin, ellagitannins, punicalagin, and ellagic acid) interact with all the five gene products under investigation. In the in vivo study, rheumatoid arthritis (RA) was induced in female Black 6 mice, and different doses of pomegranate peel extract (PPEx) (100 mg/kg and 200 mg/kg) were tested to assess their effects on healing. The evaluation of PPEx on wound healing, using the flow cytometry Annexin V test, indicated that the 200 mg/kg PPEx treatment could regulate the balance between early and late apoptosis and tissue proliferation. Furthermore, PPEx significantly accelerated wound closure rates and reduced the expression of inflammatory markers (TNF-α and IL-6) as well as transcription factors (NF-κB p65, RORγt, and T-bet), and the degree of reduction increased significantly with higher concentrations of PPEx. In conclusion, our findings suggest that PPEx has the potential as a natural therapeutic agent to enhance wound healing in RA patients by reducing inflammation and promoting balanced apoptosis.