Hepatoprotective Activity of Caesalpinia pulcherrima Extract against Paracetamol-induced Cytotoxicity: An In vitro Investigation

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Abstract
Introduction and Aim: The study aimed to explore the hepatoprotective potential of the methanolic extract of Caesalpinia pulcherrima L. (MECP) using the paracetamol-induced hepatotoxicity model in HepGⁿ cells. Methods: The MECP was analysed by GC-MS to identify its major components. In silico studies were performed using pro-inflammatory markers, including IL-6 and TNF-α. Pre-treated HepG2 cells (6.25-100 μg/mL) were exposed to paracetamol (40 mM, 12 h) after 24 hours. LDH, oxidative markers (SOD, GPx, GSH, MDA) were quantified, and the levels of inflammatory cytokines (TNF-α, IL-6) were measured after MECP treatment. Cellular morphology was analysed by fluorescence microscopy to assess the effect of MECP on cell growth. Results: GC-MS spectra showed the presence of 167 compounds in the MECP extract. An insilico study identified thymol and catechol as potent molecules with promising binding to IL-6 and TNF-α, supporting the traditional claim that C. pulcherrima is an anti-inflammatory compound effective in liver disorders. Paracetamol exposure significantly elevated LDH release, MDA levels, and inflammatory cytokines TNF-α and IL-6, while markedly reducing antioxidant defences, SOD, GPx, and GSH. Discussion: Pre-treatment with MECP showed restoration of the levels of these parameters toward normal in a dose-dependent manner. The 100 μg/mL MECP showed significant reductions in LDH, MDA, TNF-α, and IL-6. Microscopic characteristics confirmed cellular regeneration comparable to the control group. Thymol and catechol exhibited strong binding with IL-6 and TNF-α, suggesting their involvement in MECP’s hepatoprotective effect. Conclusion: These results support its traditional use in liver disorders and highlight its potential as a natural hepatoprotective agent.