Phytochemical profiling and in vivo evaluation of the analgesic and anti-inflammatory effects of Vitex agnus-castus L. in Wistar rats

Abstract

ABSTRACT Inflammation and pain are key physiological responses to tissue injury and infection. Although NSAIDs and opioids are widely used for their treatment, prolonged use can cause severe side effects. Vitex agnus-castus L. (VAC) has long been employed in traditional medicine for its therapeutic benefits, yet its analgesic and anti-inflammatory effects remain underexplored. This study aimed to identify the bioactive constituents of the methanolic leaf extract of V. agnus-castus from Algeria using LC-MS/MS and to evaluate its analgesic and anti-inflammatory properties in vivo. Phytochemical profiling was conducted using an Agilent LC-MS/MS system. Analgesic activity was assessed via the hot plate test, while anti-inflammatory potential was evaluated using the formalin-induced paw edema model in Wistar rats. Paracetamol and diclofenac sodium were used as reference drugs. LC-MS/MS analysis identified 14 compounds, mainly phenolic acids and flavonoids, including vanillic acid, syringic acid, isoquercitrin, rutin, and baicalin. VAC extract (100 and 200mg/kg) showed significant dose-dependent analgesic and anti-inflammatory effects. At 200mg/kg, VAC achieved 56.20% edema inhibition, comparable to diclofenac (55.74%). The methanolic extract of V. agnus-castus exhibits potent analgesic and anti-inflammatory activities, likely due to its polyphenolic and flavonoid composition, supporting its traditional medicinal use in folk medicine.