Bioactive Metabolites from Aerial Parts of Plantago indica L.: Structural Elucidation and Integrated In Vitro/In Vivo Assessment of Anti-Inflammatory and Wound-Healing Efficacy

The genus <i>Plantago</i> (Plantaginaceae) is widely distributed worldwide. The <i>Plantago</i> species are used in traditional medicine as wound healers, anti-inflammatory agents, antipyretics, and analgesics. This study aimed to investigate the phytochemical composition from the aerial parts of <i>Plantago indica</i> L. and to evaluate its biological activities. Isolation studies and in vitro investigations were conducted on an aqueous phase of 80% EtOH extract of <i>Plantago indica</i>. In addition, in vivo studies were carried out using the MeOH, 80% EtOH, and water extracts. Plantarenaloside (<b>1</b>), 3-oxo-<i>α</i>-ionol <i>β</i>-glucoside (<b>2</b>), martynoside (<b>3</b>), acteoside (<b>4</b>), feruloyl gardoside (<b>5</b>), and ursolic acid (<b>6</b>) were isolated from the extract. The structures of the compounds were elucidated using 1D- and 2D-NMR and ESI-MS analyses. The extract, fractions, and pure compounds were tested in vitro for cytotoxicity (MTT), anti-inflammatory activity (NO, IL-6, and TNF-<i>α</i> production), wound healing (scratch test), and antioxidant capacity (DPPH, ABTS, SO). Feruloyl gardoside (20.11-58.27%) significantly reduced NO levels at concentrations of 25-100 µM. It significantly reduced IL-6 levels (40.17%) at 100 µM. Additionally, the in vivo anti-inflammatory (acetic acid-induced vascular permeability) and wound healing (incision and excision models) effects of the extracts were investigated. The findings suggest that <i>P. indica</i> may be considered to be a potential therapeutic option for managing inflammation and for promoting wound healing.