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

Abstract

BACKGROUND: Non-alcoholic fatty pancreas (NAFP) is characterized by abnormal pancreatic fat accumulation and remains poorly understood. AIM: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.