Dual calcium-ACE inhibition reverses non alcoholic fatty pancreas progression: Mechanistic convergence on miR-762/GPX4 driven NLRP3 inflammasome activation

Non-alcoholic fatty pancreas (NAFP) is characterized by abnormal pancreatic fat accumulation and remains poorly understood. This study investigates the therapeutic potential and mechanistic basis of low-dose amlodipine-perindopril combination therapy, hypothesizing dual calcium channel blockade and RAS inhibition synergistically disrupt ferroptotic-inflammatory cascades driving pancreatic lipotoxicity. The bioinformatics analysis of GEO datasets (GSE40901, GSE101462, and GSE194331) identified differentially expressed genes, which were further validated through molecular docking studies with amlodipine and perindoprilat. Male Wistar rats (n = 30) were randomized into five groups: healthy control, NAFP (high-sugar/high-fat diet), amlodipine-treated, perindopril-treated, and combination therapy. Gene expression of GPX4, ACSL4, NLRP3, and rno-miR-762 was quantified by Real-Time PCR. Metabolic parameters, pancreatic histopathology, and immunohistochemical markers (caspase-1, nuclear factor kappa B (NF-κB)) were assessed. Bioinformatics analysis revealed significant alterations in ferroptosis and inflammatory pathways. Molecular docking demonstrated favorable binding affinities between therapeutic compounds and target proteins. NAFP induction resulted in GPX4 downregulation (0.471 ± 0.357 vs. 1.197 ± 0.449, P < 0.05), ACSL4 upregulation (48.633 ± 40.998 vs. 1.197 ± 0.445, p < 0.001), elevated NLRP3 (12.717 ± 7.904 vs. 1.115 ± 0.186, p < 0.001), and increased rno-miR-762 expression. The combination therapy achieved 73 % reduction in pancreatic adipocyte area (vs. 42 % with monotherapies, p < 0.001) through coordinated miR-762 suppression (↓68 %, q=0.003) and GPX4 protein restoration (↑2.1-fold, p = 0.008). NLRP3 inflammasome activity decreased by 89 % (p = 0.004). Crucially, the regimen preserved hemodynamic stability, demonstrating metabolic benefits without cardiovascular compromise. These findings position miR-762 as a linchpin biomarker connecting metabolic stress to ferroptotic pancreatitis, with clinical translation challenges. This work provides the first evidence that sub-therapeutic calcium-RAS crosstalk modulation can reverse established pancreatic steatosis while outlining a precision medicine framework for NAFP prevention. • Low-dose amlodipine–perindopril reverses diet-induced non-alcoholic fatty pancreas. • Combination therapy restores GPX4 and suppresses ACSL4-driven ferroptosis. • miR-762 acts as a key regulator linking metabolic stress to fatty pancreas disease. • Dual calcium–RAS blockade attenuates NLRP3 inflammasome activation.