Liver toxicity of macrolide antibiotics in zebrafish
Miao-Qing Zhang1, Bo Chen2, Jing-Pu Zhang2
1Postdoctoral Scientific Research Workstation, China Resources Sanjiu Medical & Pharmaceutical Co., Ltd., Shenzhen 518110, China; Postdoctoral Mobile Research Station, State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences & School of Chemical Engineering, University of Chinese Academy of Sciences, Beijing 100190, China; Shenzhen China Resources Gosun Pharmaceuticals Co., Ltd., Shenzhen 518049, China; Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Abstract:
Macrolide antibiotics (macrolides) are among the most commonly prescribed antibiotics worldwide and are used for a wide range of infections, but macrolides also expose people to the risk of adverse events include hepatotoxicity. Here, we report the liver toxicity of macrolides with different structures in zebrafish. The absorption, distribution, metabolism, excretion and toxicology (ADMET) parameters of macrolide compounds were predicted and contrasted by utilizing in silico analysis. Fluorescence imaging and Oil Red O stain assays showed all the tested macrolide drugs induced liver degeneration, changed liver size and liver steatosis in larval zebrafish. Through RNA-seq analysis, we found seven co-regulated differentially expressed genes (co-DEGs) associated with metabolism, apoptosis and immune system biological processes, and two co-regulated significant pathways including amino sugar and nucleotide sugar metabolism and apoptosis signaling pathway. We found that only fosab of seven co-DEGs was in the two co-regulated significant pathways. fosab encoded proto-oncogene c-Fos, which was closely associated with liver diseases. The whole-mount in situ hybridization showed high transcription of c-Fos induced by macrolide compounds mainly in the liver region of zebrafish larvae. Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) leakage assays revealed that macrolides exerts significant cytotoxic effects on L02 cells. qRT-PCR and western blot analysis demonstrated macrolides also promoted human c-Fos expression in L02 cells. The c-Fos overexpression significantly reduced cell viability by using CCK-8 assay. These data indicate that hepatotoxicity induced by macrolides may be correlated with c-Fos expression activated by these compounds. This study may provide a biomarker for the further investigations on the mechanism of hepatotoxicity induced by macrolide drugs with different structures, and extend our understanding for improving rational clinical application of macrolides.
Insights
Macrolide antibiotics can cause liver damage. This study reveals macrolides induce liver toxicity in zebrafish, linked to increased c-Fos expression, potentially serving as a biomarker for drug safety.
Area of Science:
- Pharmacology and Toxicology
- Molecular Biology
- Zebrafish Models
Background:
- Macrolide antibiotics are widely used but associated with hepatotoxicity.
- Understanding the mechanisms of macrolide-induced liver injury is crucial for patient safety.
Purpose of the Study:
- To investigate the hepatotoxicity of structurally diverse macrolide antibiotics in zebrafish.
- To identify molecular mechanisms underlying macrolide-induced liver injury, focusing on gene expression changes and potential biomarkers.
Main Methods:
- In silico prediction of ADMET parameters for macrolide compounds.
- Zebrafish models for liver toxicity assessment (fluorescence imaging, Oil Red O staining).
- RNA-sequencing (RNA-seq) for gene expression analysis, qRT-PCR, Western blot, and cell-based assays (CCK-8, LDH) in L02 cells.
Main Results:
- Macrolides induced liver degeneration, altered liver size, and steatosis in zebrafish larvae.
- RNA-seq identified shared molecular pathways and differentially expressed genes, including proto-oncogene c-Fos.
- Macrolides increased c-Fos expression in zebrafish liver and human L02 cells, correlating with reduced cell viability.
Conclusions:
- Hepatotoxicity of macrolides may be linked to c-Fos activation.
- c-Fos could serve as a potential biomarker for investigating macrolide-induced liver injury.
- Findings contribute to a better understanding of macrolide drug safety and clinical application.
