Mg & sup2;(+) deficiency has been linked to oxidative stress, inflammation, and hypertension. We have previously shown that Losartan increases plasma Mg & sup2;(+) levels in Dahl salt-sensitive (Dahl) rats, suggesting that defective Mg & sup2;(+) metabolism in this strain contributes to hypertension. Here, we investigated the effects of dietary Mg & sup2;(+) supplementation (0.3% MgO) and/or Losartan (10 mg/kg/day, in drinking water) in male Dahl rats fed a high-NaCl (8%) or standard Na+ diet for six weeks. Six-week-old rats were randomly assigned to eight experimental groups. NO metabolites (nitrites and nitrates) were measured in plasma and kidney homogenates. Oxidative stress was assessed by lipid peroxidation in plasma and red blood cell ghosts (RBCG), and by erythrocyte osmotic fragility (OS50). Both Losartan and Mg & sup2;(+) supplementation protected against hypertension and high-salt-induced systemic and local oxidative stress, with differential regulation of Na+/K+-ATPase and Na+-ATPase activity. A consistent "high Na+ effect" was observed across NaCl-fed groups, characterized by Renin-Angiotensin-Aldosterone System (RAAS) activation, increased Reactive Oxygen Species (ROS) production, lipid peroxidation, osmotic fragility, and elevated Na+/K+-ATPase activity. Losartan normalized blood pressure (BP) by attenuating renal RAAS activity and reducing oxidative stress, whereas Mg & sup2;(+) supplementation increased (Nitrites + Nitrates) NOX levels, improved red blood cell (RBC) membrane stability, and stimulated Na+-ATPase activity. These findings highlight distinct but complementary mechanisms through which AT1R blockade and Mg & sup2;(+) supplementation attenuate salt-induced oxidative stress and reduce BP. Future studies incorporating immunohistochemistry and qPCR are warranted to further characterize ROS-mediated renal tissue damage across treatment groups.
Anemia, characterized by decreased hemoglobin levels and impaired oxygen-carrying capacity, remains a major global public health concern. Conventional therapies, such as blood transfusions and oral iron supplementation, are often associated with adverse effects and limited absorption, highlighting the need for safer and more effective alternatives. Natural products rich in polyphenols and essential minerals have attracted increasing attention due to their multifunctional therapeutic potential. In this context, the present study is the first to investigate the anti-anemic potential of Pinus pinaster Aiton and Pinus halepensis Mill. bark extracts through an integrated phytochemical, nutritional, in vivo, in vitro, and in silico approach. Phytochemical profiling by HPLC-MS/MS revealed several major phenolic compounds, including quercetin, catechin, protocatechuic acid, and taxifolin-3-O-glucoside. The extracts exhibited high total polyphenol and flavonoid contents and strong antioxidant activity, as demonstrated by DPPH, ABTS, PR, and TAC assays. Mineral analysis revealed significant levels of essential elements, including iron, magnesium, potassium, sodium, and zinc. In phenylhydrazine-induced anemic Wistar rats, 21-day treatment with the extracts significantly restored hematological parameters (RBC, Hb, and HCT), serum iron and ferritin levels, and improved renal and hepatic biomarkers compared with untreated anemic controls. In vitro assays further demonstrated anti-hemolytic activity and enhanced erythrocyte membrane stability. Molecular docking analysis supported these findings by revealing favorable interactions between the major phenolic compounds and key targets involved in erythropoiesis, iron metabolism, and oxidative stress, including the erythropoietin receptor, superoxide dismutase, and ferrochelatase. Collectively, these findings highlight the potent anti-anemic and protective effects of Pinus bark extracts and suggest their potential as promising natural therapeutic agents.
