Hepatic GSTM3 deficiency Accelerates Atherosclerosis through redox-driven mitochondrial dysfunction and necroinflammation in Apoe-/- mice.

BACKGROUND AND AIMS: Metabolic dysfunction-associated steatotic liver disease (MASLD) is closely associated with accelerated atherosclerosis and cardiovascular complications, yet the hepatic redox-dependent mechanisms linking cholesterol overload to systemic vascular risk remain poorly defined. Glutathione S-transferase mu 3 (GSTM3) is a lipid peroxide-detoxifying enzyme within the hepatic antioxidant defense system, but its role in oxidative stress-induced dyslipidemia and associated atherogenesis is unknown.

METHODS

In this study we integrated GSTM3-interactome profiling, hepatocyte-specific GSTM3 silencing, and in vivo GSTM3 knockdown in hypercholesterolemic Apoe-/- mice to determine if GSTM3 regulates hepatic redox balance, mitochondrial homeostasis, and atherosclerosis progression under cholesterol overload.

RESULTS

High-cholesterol feeding induced a biphasic hepatic GSTM3 response followed by depletion during sustained injury, coinciding with heightened steatohepatitis and plaque formation. GSTM3 knockdown in hepatocytes amplified cholesterol-induced oxidative stress, disrupted mitochondrial biogenesis and dynamics, and promoted necroinflammatory cell death. In Apoe-/- mice, hepatic GSTM3 loss markedly worsened glutathione depletion (∼55%, p < 0.0001) and lipid peroxidation (∼2-fold, p < 0.0001), with enhanced fibrogenesis and inflammasome activation. These hepatic alterations were associated with a more atherogenic lipid profile and significantly increased aortic plaque burden (∼24.8%, p < 0.01) compared with hypercholesterolemic controls.

CONCLUSION

Hepatic GSTM3 functions as a redox gatekeeper that preserves mitochondrial integrity and restrains necroinflammation during cholesterol overload. Loss of GSTM3 drives redox-induced mitochondrial dysfunction and systemic oxidative dysregulation, thereby accelerating atherogenesis. These findings identify hepatic redox buffering capacity as a determinant of cardiovascular risk in metabolic disease.