Pharmacological evaluation of Adenostemma lavenia acetone extract in Swiss Albino mice: Analgesic, anti-inflammatory, and thrombolytic insights from in vivo, in vitro, density functional theory, and molecular docking studies

Abstract

Background: Adenostemma lavenia, a plant long used in traditional medicine to manage pain, inflammation, and circulatory disorders, has not been thoroughly validated using modern scientific methods. This study presents the first comprehensive investigation into the analgesic, anti-inflammatory, and thrombolytic potentials of the acetone extract of A. lavenia leaves (AEAL), employing an integrated strategy that combines in vivo, in vitro, and in silico methodologies.

Methods: The analgesic effects of the extract AEAL were evaluated in mice (five mice per group) using acetic acid-induced writhing, formalin-induced pain, and hot plate tests. Anti-inflammatory activity was assessed through xylene-induced ear edema in mice, heat-induced hemolysis of human red blood cells (HRBC), and protein denaturation assays. In vitro clot lysis was performed in triplicate to determine the thrombolytic activity of AEAL and a standard streptokinase. Molecular docking, density functional theory (DFT), and ADME/T profiling were conducted to identify and evaluate potential bioactive compounds.

Results: AEAL demonstrated significant, dose-dependent analgesic effects, reducing writhing responses by 53.74% (at 400 mg/kg), and inhibiting formalin-induced pain by up to 65.76% (p < 0.01). It also showed potent anti-inflammatory activity, with edema inhibition of 55.17% at a 400 mg/kg dose (p < 0.001), HRBC membrane stabilization (77.19%), and 85.96% suppression of protein denaturation. In thrombolytic testing, AEAL significantly (p < 0.001) achieved 68.1% clot dissolution compared with the control group. In silico analysis revealed that sesquiphellandrene strongly bound to COX-2 (−7.7 kcal/mol) and tissue plasminogen activator (tPA) (−6.9 kcal/mol), whereas derivatives of 2,3-hexadienoic acid exhibited notable COX-2 inhibition (−7.4 kcal/mol). ADMET predictions indicated favorable drug-like properties. In the DFT analysis, sesquiphellandrene and 2,3-hexadienoic acid, 2-methyl-4-phenyl-, methyl ester exhibited a narrow HOMO–LUMO energy gap, indicating high chemical reactivity and a greater propensity for electron-transfer processes.

Conclusions: The AEAL exhibits robust analgesic, anti-inflammatory, and thrombolytic activities, corroborated by computational analysis of its phytoconstituents. These results validate its ethnopharmacological use and highlight sesquiphellandrene as a promising candidate for future therapeutic development. Further isolation, mechanistic studies, and preclinical evaluation of sesquiphellandrene are warranted to harness its full pharmacological potential.

Keywords

Adenostemma lavenia / ADME/T / analgesic / DFT / HOMO–LUMO / in silico

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