Abstract

Aluminum (Al) is a toxic heavy metal that stores in the spleen, where it induces structural damage and disrupts immune function. Hesperidin, a citrus derived flavonoid, is known for its strong antioxidant activities. The present study evaluated the protective role of hesperidin against aluminum chloride (AlCl3) induced splenic damage. Forty Male Wistar rats were classified to 4 groups: control, hesperidin alone (100 mg/kg), AlCl? alone (128 mg/kg), and combined AlCl? plus hesperidin, all administered orally for 30 days. Splenic tissues were assessed for oxidative stress biomarkers (MDA, GSH, SOD, CAT), Nrf2/HO 1 antioxidant pathway, necroptosis and apoptosis markers (RIPK1, RIPK3, MLKL, caspase 3, Bax, Bcl 2), TNF ? expression, and histopathological alterations. AlCl? exposure significantly increased (MDA) and suppressed antioxidant defenses (SOD, CAT, GSH). It also downregulated Nrf2/HO 1 signaling, elevated TNF ?, activated necroptotic mediators (RIPK1, RIPK3, MLKL), and triggered apoptosis, by increased caspase 3 and Bax with reduced Bcl 2. Histologically, AlCl? caused marked lymphocyte depletion and expansion of the red pulp. Co administration of hesperidin markedly ameliorated these changes by enhancing Nrf2/HO 1 activation, restoring antioxidant enzyme levels, reducing TNF ? expression, suppressing necroptotic and apoptotic pathways, and improving splenic architecture. In conclusion, hesperidin protects against AlCl? induced splenic injury through dual mechanisms: activation of the Nrf2/HO 1 antioxidant axis and inhibition of TNF ? driven necroptosis and apoptosis. These findings highlight hesperidin as a promising dietary supplement for mitigating aluminum related immunotoxicity.

 Keywords: Hesperidin, aluminum chloride, spleen, necroptosis, inflammation, oxidative stress