Sex-specific mitigation of oxidative and inflammatory cardiac alterations by folic acid and simvastatin in chronic mild hyperhomocysteinemia.

Mild hyperhomocysteinemia (HHcy), characterized by plasma homocysteine levels of 16-30 μmol/L, induces oxidative stress and inflammation in cardiovascular tissues. Folic acid (FA) and simvastatin (SIM) possess antioxidant and anti-inflammatory properties. We investigated their acute cardioprotective effects in hearts exposed to chronic mild HHcy. Twelve-month-old Wistar rats received saline (control) or DL-homocysteine (0.03 μmol/g, sc, twice daily) for 30 days. Post-mortem, 300 μm heart slices were incubated ex vivo with FA (100 μM, 60 min) or SIM (10 or 30 μM, 80 min). Oxidative stress and antioxidant status were assessed by ROS, TBARS, sulfhydryl content, nitrites, and CAT, SOD, and GPx activities. NRF2 protein and Nfe2l2 expression evaluated antioxidant signaling, while cytosolic/nuclear NFκB p65 and IL-1α, IL-1β, IL-6, IL-10, and RELA mRNA assessed inflammatory responses. In males, HHcy increased ROS, TBARS, CAT, cytosolic NFκB p65, and IL-1α/IL-1β/IL-6, while reducing IL-10 and NRF2; FA and SIM reversed these changes, with SIM (10 μM) uniquely enhancing nitrites, SOD/GPx, and Nfe2l2. In females, HHcy decreased GPx and nitrites and increased IL-6 and RELA; FA and SIM normalized these, with SIM strongly activating SOD, CAT, GPx, and cytosolic NRF2 even in controls. These findings demonstrate that FA and SIM provide acute cardioprotection under chronic HHcy by restoring redox balance and attenuating inflammatory signaling in a sex-dependent manner, highlighting SIM's pleiotropic enhancement of NRF2-mediated antioxidant defenses and nitric oxide bioavailability.