Agathisflavone Modulates the Kynurenine Pathway and Glial Inflammatory Responses with Implications for Neuroprotection

The cells in the central nervous system (CNS) can adapt to injury and inflammation through structural and functional changes, many of which are mediated by the kynurenine pathway (KP). Studies using glia-neuron co-cultures showed that the biflavonoid agathisflavone (FAB), purified from the leaves of <i>Cenostigma pyramidale</i> Tul., a plant native to the Brazilian caatinga, exerts strong neuroprotective effects. This study evaluated whether agathisflavone (1 µM) modulates these responses in human and murine astrocytes and microglia exposed to inflammatory activation with lipopolysaccharide (LPS, 1 µg/mL), excitotoxic activation of NMDA receptors with quinolinic acid (QUIN, 500 µM), or inhibition of the KP rate-limiting enzyme indoleamine 2,3-dioxygenase 1 (IDO1) with 1-methyl tryptophan (1-MT, 1.5 μM). Co-treatment with FAB increased astrocyte viability relative to LPS, QUIN, or 1-MT alone, by up to 35% (<i>p</i> < 0.05), while reducing GFAP overexpression and other features of reactive astrogliosis. FAB decreased the proportion of Iba-1⁺ microglia, indicating anti-inflammatory effects. When combined with QUIN or 1-MT, FAB reversed the elevation of iNOS (<i>p</i> < 0.0001) and reduced IL1β upregulation. FAB also modulated KP activity in a cell type-specific manner. In astrocytes, FAB with QUIN or with 1-MT increased IDO activity, whereas in microglia, FAB alone reduced it. In microglia, kynurenine-3-monooxygenase (KMO) expression was significantly increased under FAB+QUIN or FAB+1-MT (<i>p</i> < 0.0001). Finally, astrocyte-conditioned medium from FAB-treated cells increased the viability of neuron-like PC12 cells by up to 40%. Collectively, these findings show that FAB confers cytoprotective and anti-inflammatory actions on glial cells, modulates KP signalling in a context-dependent manner, and supports neuronal survival under neuroinflammatory conditions.