Hepatoprotective Mechanism of Shorea Robusta Gaertn. f. Against Isoniazid-induced Hepatotoxicity by Targeting TNF-α, TGF-β1, and PPAR-α: Insights from Computational Profiling To Immunohistochemical Confirmation.

Shorea robusta (Dipterocarpaceae), a timber-producing tree, that thrives in India, Bangladesh, East Himalaya, Nepal, Tibet, and West Himalaya regions. Traditionally, different parts of this plant have been used for several pharmacological activities. The present study was aimed to validate the hepatoprotective mechanisms of Shorea robusta in in silico, in vitro and in vivo model. The methanolic extract of Shorea robusta (MESR) bark was analyzed using UHPLC-Q-TOF-MS, revealing 68 numbers of phytoconstituents. Based on Lipinski’s Rule of Five and ADMET analysis, 15 major phytoconstituents were selected for molecular docking studies against hepatoprotective targets such as TNF-α, TGF-β1, and PPAR-α. In vitro antioxidant study was evaluated using DPPH and ABTS assays. Hepatoprotective efficacy was assessed in an isoniazid (INH)-induced hepatotoxicity rat model through biochemical parameters, cytokine analysis, histopathology, and immunohistochemistry. Molecular docking analysis revealed that Lysidiside N (-11.0 kcal/mol), polydatin (-10. Kcal/mol), resveratrol 12-C-beta-glucopyranoside (-10. Kcal/mol), alpinetin (-9.2 Kcal/mol), and ellagic acid (-9.1 Kcal/mol) displayed the highest docking score toward TNF-α, TGF-β1, and PPAR-α. MESR exhibited high total phenolic (29.922 mg GAE/g D.W.) and flavonoid content (29.74 mg QE/g D.W.), with strong antioxidant properties (DPPH: IC50 = 39.12 µg/mL; ABTS: IC50 = 37.68 µg/mL). Oral administration of MESR (500 mg/kg) was significantly restored altered liver enzymes, protein, and lipid profiles, along with cytokine levels. Histopathological and immunohistological analyses revealed marked improvement in the liver architecture, indicating enhanced hepatoprotection in the treated group. Collectively, MESR at a dose of 500 mg/kg exhibited a pronounced hepatoprotective activity against INH-induced hepatotoxicity in experimental rats. Lysidiside N, Polydatin, resveratrol 12-C-β-glucopyranoside, alpinetin, and ellagic acid may be considered as the major phytoconstituents contributing to the hepatoprotective activity of S. robusta bark.