RNAseqencing of wild type (WT) and Ripk3-/- (KO) AML-12 cells in the presence (PA) of absence (C) of 125 µM palmitate-bovine serum albumin for 24 h

Purpose: Necroptosis as been implicated in various deseases. The goal of this study is to invastigate the impact of RIPK3 in the lipid metabolism of hepatic cells. Methods: AML-12 cells invalidated or not for RIPK3 were exposed to free fatty acid and the mRNA profiles of wild type (WT) or knockout (RIPK3-KO) AML-12 cells control (C) or exposed to palmitic acid (PA) were generated by deep sequencing, in 5 copies, using Illumina NOVAseq 6000 plateform. Differential expression analysis between two conditions/groups (five biological replicates per condition) was performed using DESeq2 R package. Genes with an adjusted P value < 0.05 found by DESeq2 were assigned as differentially expressed. qRT–PCR validation was performed using SYBR Green assays Results: The DEGs were clustered using a hierarchical clustering algorithm, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis unveiled that transcripts associated with OXPHOS and thermogenesis were markedly upregulated in unchallenged Ripk3-/- hepatocytes compared with WT cells an effect that was aggravated by the PA treatment. Similarly, gene ontology enrichment analysis showed that mitochondria respiration-related proteins were overrepresented among the cellular components that were upregulated in Ripk3-/- hepatocytes compared with WT cells Conclusions: The absence of Ripk3 impacts on hepatocyte transcriptomic profile, increasing the expression of genes implicated in mitochondria bioenergetics and function, rescuing mitochondria respiration after free fatty acid overload. RNAseqencing of wild type (WT) and Ripk3-/- (KO) AML-12 cells in the presence (PA) of absence (C) of 125 µM palmitate-bovine serum albumin for 24 h