Background: As a major cause of liver disease worldwide, metabolic associated fatty liver disease (MAFLD) comprises non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH). Due to the high prevalence and poor prognosis of NASH, it is critical to understand its mechanisms. However, the etiology and mechanisms remain largely unknown.
Methods: We first identified differential genes in NASH by single-cell analysis of the GSE129516 and expression profiling data analysis of the GSE184019 dataset from the GEO database. Then single-cell trajectory reconstruction and analysis, immune gene score, cellular communication, key Gene Screening, functional enrichment analysis, and immune microenvironment analysis were carried out. Finally, cell experiments were performed to verify the role of key genes in NASH.
Results: We show transcriptome profiling of 30,038 single cells—including hepatocyte and non-hepatocyte—from normal and steatosis adult mouse liver. Comparative analysis of hepatocyte and non-hepatocyte revealed pronounced heterogeneity as well as specific utilization of non-hepatocyte types as major cell-communication hubs. The results showed that Hspa1b, Tfrc, Hmox1 and Map4k4 effectively distinguish NASH tissues from normal samples. The results of scRNA-seq and qPCR indicated that the expression levels of hub genes in NASH were significantly higher than in normal cells or tissues. Further immune infiltration analysis showed significant differences in Macrophages M2 distribution between healthy and metabolic associated fatty liver samples.
Conclusions: Our results suggest that Hspa1b, Tfrc, Hmox1 and Map4k4 are all potential biomarkers for NASH diagnosis and prognosis may also be potential therapeutic targets for NASH.