In Vitro and In Vivo Studies on the extracts of Vitex Negundo and Hygrophilla Auriculata for their Hepatoprotective activity

This study aimed to evaluate the hepatoprotective potential of aqueous and methanolic extracts of Vitex negundo and Hygrophila auriculata using in-vitro and in-vivo models. Phytochemical analysis revealed that V. negundo contains carbohydrates, glycosides, tannins, and saponins, while H. auriculata contains carbohydrates, alkaloids, glycosides, and proteins—compounds known for their therapeutic properties. Antioxidant activity was assessed using DPPH and Nitric Oxide radical inhibition assays. The methanolic extract of V. negundo demonstrated the strongest DPPH radical scavenging activity with an IC₅₀ of 28.1 ± 0.425 µg/ml, while the methanolic extract of H. auriculata showed the highest nitric oxide scavenging activity with an IC₅₀ of 117.55 ± 0.6708 µg/ml. These results were compared to standard antioxidants like Ascorbic acid and Rutin. In-vitro cytoprotective studies were carried out on BRL-3A liver cells subjected to oxidative stress. The aqueous and methanolic extracts of V. negundo provided 50% cytoprotection at 140 µg/ml and 570 µg/ml, respectively, whereas H. auriculata extracts achieved similar protection at 220 µg/ml (aqueous) and 337.5 µg/ml (methanolic). Hepatoprotective activity was further evaluated in isolated rat hepatocytes exposed to D-Galactosamine-induced toxicity. All extracts significantly restored biochemical markers such as liver enzymes, total protein, and albumin. Among the extracts, the aqueous extract of H. auriculata showed the most potent hepatoprotective effect. In-vivo studies in rats confirmed these findings. Rats treated with D-GalN alone showed elevated liver enzymes and markers of liver damage, while co-treatment with plant extracts significantly reversed these changes. The aqueous extract of H. auriculata again showed the most notable hepatoprotection, followed by its methanolic extract. Extracts of V. negundo also demonstrated hepatoprotective effects, though to a lesser extent. Histopathological examination of liver tissues supported the biochemical results, showing reduced liver damage in extract-treated groups. Overall, the findings highlight the significant antioxidant and hepatoprotective activities of both plants, particularly the aqueous extract of H. auriculata, suggesting their potential for liver disorder therapies.