Chemical Fingerprinting, Anti-Inflammatory, and Antioxidant Potential of Hydroethanolic Extract of Aesculus indica

Aesculus indica is a remarkable species from Sapindaceae family, traditionally used for the treatment of various ailments due to the presence of a variety of bioactive compounds. The present study was planned to evaluate the chemical characterization, anti-inflammatory and antioxidant potential of hydroethanolic extract of A. indica using in vitro and in vivo approaches. A. indica fruit was extracted with a hydroethanolic (70% v/v) solution, filtered, concentrated on a rotary evaporator and crude extract was obtained. In vitro anti-inflammatory potential of A. indica was carried out against peripheral blood mononuclear cells (PBMCs) and a whole blood assay (WBA). Effects of A. indica extracts on proinflammatory cytokines (TNF-α, IFN-gamma, IL-6, IL-1β) and inflammatory mediators (NF-κB, NO and PGE₂) concentration in the supernatant of PBMCs and WBA were evaluated using commercial ELISA kits. In vivo anti-inflammatory potential of A. indica hydroethanolic extract was evaluated with carrageenan-induced paw edema in rats. A total of 36 different compounds (mostly phenolics) were detected in A. indica extract with high performance liquid chromatography (HPLC) and UHPCL-QTOF-MS/MS. The extract showed very low cytotoxicity with an IC₅₀ value of 483.68 μg/mL and significantly reduced the levels of proinflammatory cytokines and inflammatory mediators in both PBMCs and WBA models. Furthermore, the extract also effectively inhibited the paw edema by carrageenan in the 2nd hour at 400 mg/kg (73%). Histopathological analysis of rat paw tissue showed significant reduction of cellular infiltration and decrease in swelling of epidermis and dermis by A. indica extracts. The level of enzymatic antioxidants such as superoxide dismutase (SOD) and Catalase (CAT), lipid peroxidation like malondialdehyde (MDA), oxidative stress parameters including total antioxidant status (TAS) and total oxidant status (TOS) and myeloperoxidase (MPO) activity in rat paw tissues were significantly altered after treatment. The combined findings provide evidence that hydroethanolic extract of A. indica is a potential source of bioactive compounds with significant anti-inflammatory and antioxidant activities.