Systems Biology Approach to Unraveling Transcriptomic Mechanisms of Ganfule Capsules in Ameliorating Nonalcoholic Fatty Liver Disease.

Abstract

Aims: The primary objective of this study is to explore the impact of Ganfule (GFL), a traditional Chinese medicine, on differentially expressed genes (DEGs) linked to nonalcoholic fatty liver disease (NAFLD). By identifying potential biomarkers, we seek to enhance GFL's clinical efficacy through targeted pharmaceutical design.

Background: NAFLD a prevalent liver disorder, is often associated with obesity and metabolic syndrome. While GFL has demonstrated clinical efficacy in treating NAFLD, its precise targets and mechanisms of action remain elusive. Understanding these mechanisms could pave the way for more effective treatments.

Objectives: GFL, a long-standing traditional Chinese medicine (TCM), has demonstrated clinical effectiveness in treating NAFLD. However, its precise targets and mechanism of action remain elusive. In this study, we aim to explore GFL's impact on differentially expressed genes, which could potentially serve as biomarkers for developing targeted therapies. This approach is intended to enhance GFL's clinical efficacy by identifying key genes that respond to its treatment.

Methods: To induce NAFLD, 23 Sprague-Dawley rats were fed a high-fat diet. These rats were then categorized into three groups: normal diet (NOR), high-fat diet model (HFD), and those treated with GFL. Highthroughput sequencing was employed to identify DEGs in their livers. Utilizing the STRING and DAVID databases, we analyzed potential protein interactions expressed by these genes. Furthermore, the KEGG, Reactome, and Wiki databases aided in determining their biological roles and signaling pathways. Key DEGs' mRNA expression levels and corresponding proteins were further screened and confirmed through haematoxylin- eosin staining (HE), immunohistochemistry (IHC), Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR), and western blotting.

Results: Significant variations in DEGs were observed across the three groups, with 19 intersecting genes identified within a cluster of 90 NAFLD-related genes. GFL was found to adjust the expression of nine core DEGs, including Abcg1, Igfgb1, Lepr, Pdk4, Socs3, and Stat3. These genes-related proteins are tied to proteins such as FABP4, LEPR, SCD1, SOCS3, and STAT3, which are intimately connected to adipocytokine and adipogenesis pathways. Our study reveals that GFL modifies the expression of IGFBP1, LEPR, PDK4, SCD1, and SOCS3, thereby regulating the adipocytokine, JAK-STAT, leptin-insulin signaling, and adipogenesis metabolic pathways, respectively.

Conclusions: This study enhances understanding of GFL's efficacy and identifies potential biomarkers for NAFLD treatment. Optimizing GFL's efficacy and elucidating its mechanism provides a methodological reference for traditional Chinese medicine exploration.