Pirfenidone restores metabolic hormones and cardiac autophagy via p-AMPK in MASH

Abstract

Obesity promotes chronic inflammation and insulin resistance, disrupting key metabolic hormones (e.g., insulin and leptin) and triggering metabolic dysfunction-associated steatohepatitis (MASH). Obesity is commonly associated with cardiovascular disease. Autophagy preserves heart function in response to various stresses. Pirfenidone (PFD) is an antifibrotic and anti-inflammatory drug. However, its effects on the regulation of metabolic hormones and cardiac autophagy in MASH have not been investigated. Male C57BL/6J mice were fed a high-fat/high-carbohydrate (HFHC) diet for 16 weeks to induce obesity and MASH. From week 8, a subgroup received PFD (300 mg/kg/day) via gavage. In vitro, H9c2 cardiomyocytes were exposed to glucolipotoxicity to simulate metabolic stress. Serum metabolic hormones, cardiac histology, microarray analysis, and autophagy-related mRNA and protein levels were analyzed. PFD treatment restored metabolic hormone balance and promoted cardiac autophagy in obese MASH mice. Mechanistically, PFD upregulated p-AMPK protein levels (P < 0.05), reversing the dysregulation of autophagy-related proteins both in vivo and in glucolipotoxicity-exposed H9c2 cells. Concurrently, PFD prevented systemic insulin resistance (P < 0.001), hepatic steatosis (P < 0.001), and liver damage (ALT/AST, P < 0.05). In cardiac tissue, PFD attenuated metabolic stress-induced hypertrophy (Nppa/Nppb mRNA), inflammation (Tnf, Il6, Adgre1, and Ccl2 mRNA, P < 0.05), and fibrosis (Tgfb1, Col1a1, and Col3a1 mRNA, P < 0.05). Pirfenidone has cardioprotective effects in obesity-associated MASH by restoring metabolic hormone levels and activating autophagy via p-AMPK signaling. These findings suggest a potential translational therapeutic strategy for treating cardiovascular complications in MASH.