Hydroethanolic Soy Extract Rich in Isoflavones Protects Dystrophic Mutant Mice From Oxidative Stress and Inflammation via the ROS/NF-κB Feedback Pathway.

Soy isoflavones have been extensively studied for their beneficial effects in various diseases. Oxidative stress induces inflammation and myonecrosis in early disease stages in both mdx mice and human patients with Duchenne muscular dystrophy (DMD). This study aimed to investigate the effects of hydroethanolic soy extract (HSE) on dystrophic muscle in mdx mice (a mouse model of DMD), based on the estrogenic action of total soy isoflavones in reducing oxidative stress and inflammation. Phytochemical profiling of HSE was performed by UPLC-PDA-MS, and in silico molecular docking identified putative targets of the main isoflavones. Fourteen-day-old male mdx mice were treated orally with HSE (57 mg/kg/day, ~2 mg/kg genistein) for 45 days. Muscle function, biochemical markers, histopathology, and molecular indicators of inflammation and oxidative stress were evaluated. HSE contained multiple isoflavone classes, showing strong binding affinities for monoamine oxidase and estrogen receptors. The isoflavone-rich HSE showed no toxic effects in mdx mice and demonstrated a clear protective influence on skeletal muscle pathology. Treatment with HSE led to improved muscle integrity, as evidenced by increased muscle strength and a marked reduction in markers of muscle damage such as IgG-positive fibers and serum CK levels. Additionally, HSE effectively mitigated the inflammatory response, with significant downregulation of NF-κB, reduction of inflammatory and macrophage infiltration areas, and attenuation of oxidative stress markers. The antioxidant response was enhanced, reflected by lower ROS production, decreased lipid peroxidation (4-HNE and lipofuscin), and elevated levels of endogenous antioxidants (catalase and GSH). Moreover, HSE appeared to support muscle regeneration by increasing the proportion of centrally nucleated fibers while modulating myogenic regulatory pathways, as indicated by decreased MyoD expression. Isoflavone-rich HSE safely improved muscle integrity and function in mdx mice by attenuating oxidative stress and inflammation through modulation of the ROS/NF-κB pathway. These findings highlight the therapeutic potential of soy-derived isoflavones as a complementary approach for managing DMD.