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Updated: May 31, 2025
Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Published on: April 3, 2017
Single-Cell RNA Sequencing Reveals Macrophage Dynamics During MASH in Leptin-Deficient Rats
Xiaoming Xin1, Yaohua Ni1, Jing Wang1
1School of Pharmacy, Shanghai University of Medicine and Health Sciences, Shanghai 201318, China.
Abstract:
Macrophages play important roles in metabolic dysfunction-associated steatohepatitis (MASH), an advanced and inflammatory stage of metabolic dysfunction-associated steatotic liver disease (MASLD). In humans and mice, the cellular heterogeneity and diverse function of hepatic macrophages in MASH have been investigated by single cell RNA sequencing (scRNA-seq). However, little is known about their roles in rats. Here, we collected liver tissues at the postnatal week 16, when our previously characterized Lep rats developed MASH phenotypes. By scRNA-seq, we found an increase in the number of macrophages and endothelial cells and a decrease in that of NK and B cells. Hepatic macrophages in rats underwent a unique M1 to M2 transition without expression of the classical markers such as Arg1 and Nos2, except for Cd163. Lipid-associated macrophages (LAMs) were increased, which could be detected by the antibody against Cd63. In the microenvironment, macrophages had an increased number of interactions with hepatocytes, myofibroblasts, T cells, neutrophils, and dendritic cells, while their interaction strengths remained unchanged. Finally, the macrophage migration inhibitory factor (MIF) pathway was identified as the top upregulated cell-communication pathway in MASH. In conclusion, we dissected hepatic macrophage dynamics during MASH at single cell resolution and provided fundamental tools for the investigation of MASH in rat models.
Insights
This study reveals unique hepatic macrophage dynamics in rat models of metabolic dysfunction-associated steatohepatitis (MASH). Macrophages shift phenotypes distinctively, interacting more within the liver microenvironment during MASH progression.
Area of Science:
- Hepatology
- Immunology
- Single-cell genomics
Background:
- Metabolic dysfunction-associated steatohepatitis (MASH) involves complex hepatic immune cell roles.
- Hepatic macrophage heterogeneity and function in MASH are well-studied in humans and mice but not rats.
- Understanding rat models is crucial for MASH research due to their unique physiology.
Purpose of the Study:
- To investigate the cellular and molecular dynamics of hepatic macrophages in a rat model of MASH.
- To characterize macrophage heterogeneity and function during MASH development at single-cell resolution.
- To identify key cell-cell communication pathways involved in MASH pathogenesis in rats.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) of liver tissues from MASH model rats.
- Analysis of cell populations, macrophage phenotypes, and cell-cell interactions.
- Identification of upregulated cell-communication pathways using computational tools.
Main Results:
- scRNA-seq revealed increased macrophages and endothelial cells, with decreased NK and B cells in MASH rats.
- Rat hepatic macrophages exhibited a unique M1 to M2 transition, marked by Cd163 but not classical markers.
- Lipid-associated macrophages (LAMs) increased, and macrophage interactions with other liver cells were altered.
- The macrophage migration inhibitory factor (MIF) pathway was identified as a top upregulated pathway.
Conclusions:
- This study provides a single-cell resolution dissection of hepatic macrophage dynamics during MASH in rats.
- Rat hepatic macrophages display unique characteristics and transitions distinct from mouse and human models.
- The findings offer essential tools and insights for future MASH research using rat models.
