Effect of valine on expression of fatty acid metabolism-regulated genes PPARα and FASN in porcine primary hepatocytes
[Objective]This study aims to explore the genes related to the regulation of liver lipid metabolism by valine in pig and the regulatory pathways,providing experimental data and theoretical basis for revealing the molecular mechanism of of the regulation.[Method]The porcine primary hepatocytes derived from 5-day-old piglet(Duroc×Landrace×Large White)were treated with 0,0.1,1.0,and 10 mmol/L valine(Val)for 24 and 48 hours,respectively.The effects of different concentrations of valine on the important genes PPARα and FASN regulating lipid metabolism were detected through Western blot analysis.Potential key genes and signaling pathways involved in the regulation were screened.With GO function and KEGG pathway enrichment analysis,real-time fluorescence quantification(qPCR)technology was used to validate differentially expressed genes.[Result](1)The results showed that 10 mmol/L of valine significantly reduced the protein expression of PPARα and FASN(P＜0.05)in porcine primary hepatocytes treated with valine for 24 and 48 hours,respectively.(2)The analysis of transcriptome sequencing data showed that compared with the control group,a total of 418 differentially expressed genes were screened in the valine group,of which 181 genes were upregulated and 237 genes were downregulated.(3)GO functional analysis revealed that differentially expressed genes mainly participated in Binding,Catalytic activity,Signal transducer activity,and Transporter activity in terms of molecular functions.(4)KEGG pathway enrichment analysis revealed that differentially expressed genes were enriched in six biological processes,including metabolism,human disease,organismal systems,cellular processes,genetic information processing,and environmental information processing.Lipid metabolism was one of the main signaling pathways involved in metabolism biological processes.The five differentially expressed genes enriched in fatty acid metabolism,including FASN,SCD,FADS2,FADS1,and HSD17B12,were mainly involved in the insulin signaling pathway,AMPK signaling pathway,and PPAR signaling pathway.[Conclusion]This study showed that 10 mmol/L of valine can downregulate the protein expression of PPARα and FASN in porcine primary hepatocytes,and regulate the activity of related genes and signaling pathways in lipid metabolism.The results of this study provide a theoretical basis for further understanding the regulation of lipid metabolism by branched chain amino acids.
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