Effect of hydrolysed <i>Trifolium repens</i> L. extract on menopause induced obesity and depressive symptoms: an <i>in-vitro</i> and <i>in-silico</i> approach

This study aimed to evaluate acid-hydrolysis effects on phytoestrogen content and the efficacy of phytoestrogen-rich extract in alleviating post-menopausal symptoms (PMS). Bioactive phytoestrogens were quantified using HPTLC, and unhydrolyzed and hydrolysed extracts were assessed for oestrogenic, anti-proliferative, MAO-A inhibition, ROS quenching, and PL inhibition potential through <i>in-vitro</i> models. In-silico analysis of quantified phytoestrogens against PMS-related proteins was conducted. Hydrolysis increased phytoestrogen content significantly, particularly biochanin-A (56.48-fold), daidzein (22.2-fold), genistein (10.77-fold), and formononetin (5.26-fold). The hydrolysed extract demonstrated improved efficacy in <i>in-vitro</i> models, including reduced IC₅₀ in oestrogenic activity, decreased cell proliferation, increased MAO-A inhibition, and de-creased IC₅₀ for lipase. High binding energies of phytoestrogens against ERβ, MAO-A, and PL supported their potential in managing post-menopausal symptoms. The study scientifically validated <i>T. repens</i> for women's health at menopause, highlighting acid-hydrolysis for enhancing therapeutic activity. <i>T. repens</i> could be explored commercially as an alternative to <i>T. pratense.</i>