FerrylHb induces inflammation and cell death in grass carp (Ctenopharyngodon idella) hepatocytes

Grass carp hemorrhagic disease is a significant problem in grass carp aquaculture. It releases highly oxidizing hemoglobin (Hb) into tissues, induces rapid autooxidation, and subsequently discharges cytotoxic reactive oxygen species (ROS). However, the mechanism underlying Hb damage to the teleost remains unclear. Here, we employed ferrylHb and heme to incubate L8824 (grass carp liver) cells and quantitatively analyzed the corresponding molecular regulation using the RNA-seq method. Based on the RNA-seq analysis data, after 12 h of incubation of the L8824 cells with ferrylHb, a total of 3738 differentially expressed genes (DEGs) were identified, 1824 of which were upregulated, and 1914 were downregulated. A total of 4434 DEGs were obtained in the heme treated group, with 2227 DEGs upregulated and 2207 DEGs downregulated. KEGG enrichment analysis data revealed that the incubation of ferrylHb and heme significantly activated the pathways related to Oxidative Phosphorylation, Autophagy, Mitophagy and Protein Processing in Endoplasmic Reticulum. The genes associated with NF-kappa B, autophagy and apoptosis pathways were selected for further validation by quantitative real-time RTPCR (qRT-PCR). The results were consistent with the RNA-seq data. Taken together, the incubation of Hb and heme induced the molecular regulation of L8824, which consequently led to programmed cell death through multiple pathways.