Boron-dipyrromethene Staining May Enhance Fat Detection in the MASLD Zebrafish Model: NGS-validated lncRNA Profiling

Abstract

Background/Aim: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a serious global public health concern. Long non-coding RNAs (lncRNAs) have been identified as key contributors to MASLD pathogenesis. Zebrafish can be utilized to study the relationship between MASLD and lncRNAs because of their similarity to human genes. Oil Red O staining is a traditional method for confirming liver fatty changes; however, it has several limitations. This study aimed to evaluate the efficacy of boron-dipyrromethene (BODIPY) in detecting fatty changes in the liver. Materials and Methods: Liver tissues were collected from 30 zebrafish that were fed a BODIPY-containing high-cholesterol diet. Oil Red O and BODIPY staining were evaluated by two pathologists, and next-generation sequencing (NGS) was performed using liver tissues categorized into high fatty change (six liver tissues) and low fatty change (six liver tissues) groups. Results: BODIPY and Oil Red O staining of zebrafish liver sections correlated significantly (p=0.009). NGS identified eight differentially expressed lncRNAs with over a 10-fold difference between the high- and low-fatty acid change groups. Of these, three showed lncRNA-mRNA interaction networks linked to human disorders. Conclusion: BODIPY staining is a reliable alternative to Oil Red O staining for assessing fatty changes in MASLD zebrafish models, particularly when examining frozen liver sections.