Transcriptomics reveal a molecular signature in the progression of nonalcoholic steatohepatitis and identifies PAI‑1 and MMP‑9 as biomarkers in in vivo and in vitro studies

Abstract

Nonalcoholic steatohepatitis (NASH) is emerging as the primary driver of liver disease-induced fibrosis. The imperative need for noninvasive biomarkers to ascertain disease progression stage is evident. The present study elucidated the biological roles of hub genes that could potentially serve as diagnostic markers for NASH. Using an in vivo approach, C57BL/6J mice were subjected to a high-fat and fructose diet (HFFD) for 6, 10, 14, 18 or 22 weeks. Serological biochemical indices were assessed and liver specimens were obtained to identify potential markers linked to the NASH process, employing a comprehensive strategy that combined transcriptomic and histopathological analyses. The HFFD regimen induced hyperlipidemia, obesity and insulin resistance, progressively culminating in NASH with fibrosis over time. The transcriptomic analyses indicated temporal patterns of pivotal gene sets intricately connected to NASH progression, which encompassed processes such as glucose homeostasis, inflammatory responses, reactive oxygen species-mediated damage, lipid metabolism disruptions and the formation of fibrotic tissue. Among these genes, Serpine1 and Mmp9 demonstrated promising diagnostic potential for NASH, with their intrahepatic mRNA expression levels serving as robust indicators. Moreover, the levels of PAI-1 (encoded by the Serpine1 gene) and MMP-9 in the serum of mice demonstrated a parallel increase with the duration of HFFD intervention. In vitro experiments utilizing HepG2 cells further validated these findings, demonstrating a significant elevation in the protein expression levels of both PAI-1 and MMP-9 upon exposure to free fatty acids, in agreement with the results of the animal study. Consequently, PAI-1 and MMP-9 are promising noninvasive biomarkers for assessing the progression of NASH.

Expression of immune related genes and possible regulatory mechanisms in different stages of non-alcoholic fatty liver disease

“…By intersecting module genes, DEGs, and IRGs, we identified 28 immune-related genes that played a role in both the onset and progression of NAFLD (Figure 3C). Previous studies performed high-throughput sequencing of liver tissue from normal mice and mice at different stages of NASH, they found that compared with normal liver tissue, the expression patterns of genes in the liver tissue of model mice and the signaling pathways involved changed as the disease progressed (37). In our study, we performed a follow-up analysis to determine whether the expression patterns of these 28 genes also changed as the NAFLD progressed.…”

Extracellular Vesicles as Delivery Vehicles for Non-Coding RNAs: Potential Biomarkers for Chronic Liver Diseases

“…The gold standard for detecting liver fibrosis is the histological analysis of biopsy specimens; however, the invasive nature of the procedure and the associated risks reduce patient compliance [ 138 ]. The utility of circulating biomarkers including circulating proteins, nucleic acids, and metabolites, as well as RNA for predicting liver fibrosis, has been described in several studies, including [ 139 , 140 , 141 , 142 ]. EV-enclosed ncRNAs have also been investigated as potential biomarkers for the grading of liver fibrosis [ 143 ].…”

Diagnostic indicators and lifestyle interventions of metabolic-associated fatty liver disease

“…Recent studies have found that as liver pathology progresses toward nonalcoholic steatohepatitis (NASH), there is a corresponding increase in the level of mRNA expression of Serpine1, as well as the protein level of PAI-1. Use of Serum PAI-1 Levels as a noninvasive biomarker to identify NASH-associated fibrosis ( 49 ). However, no experiments have been performed to assess its diagnostic value in MAFLD.…”