Single-cell RNA sequencing analyses the mechanism of scar-associated macrophages promoting hepatic stellate cell activation

doi:10.3969/j.issn.1006-7795.2024.02.015

Abstract

Abstract: Objective To establish an experimental protocol for inducing human macrophages into scar-associated macrophages (SAMs) in vitro. To investigate the mechanism underlying SAMs regulating hepatic stellate cell (HSC) activation. Methods Published single-cell RNA sequencing (scRNA-Seq) data, which were obtained from human or murine fibrotic liver tissues, were used to study the expression of fibrosis-related genes in SAMs. Phorbol-12-myristate-13-acetate (PMA) and plasminogen (PLG) were used to induce the transformation of human monocyte THP-1 into SAMs. SAM supernatant was collected to culture with LX-2. The expressions of SAM markers, fibrosis-related genes and HSC activation markers were detected by real-time quantitative polymerase chain reaction（RT-qPCR）. Results scRNA-seq revealed that SAMs were detected in both mouse fibrotic livers or human injured liver. SAMs highly expressed fibrosis-related genes such as SPP1, CTSD. PMA and PLG increased the expressions of SAM markers CD9, TRME2, LGALS3, CD63 in THP-1. Fibrosis-related genes SPP1, CTSD were highly expressed by SAMs. The supernatant of SAMs activated LX-2. Conclusion SAMs are accumulated in mouse or human liver injury/fibrosis caused by different causes, and represent similar characteristics and functions. PMA and PLG induce the transformation of human monocyte THP-1 into SAMs. SAMs promote the activation of HSCs via SPP1 and CTSD, thus promoting the occurrence and development of liver fibrosis.

Key words: single-cell RNA sequencing, scar-associated macrophage, hepatic stellate cell, liver fibrosis

CLC Number:

R575.2

iu Chang, Li Liying, Chang Na. Single-cell RNA sequencing analyses the mechanism of scar-associated macrophages promoting hepatic stellate cell activation[J]. Journal of Capital Medical University, 2024, 45(2): 271-280.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2024.02.015

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2024/V45/I2/271

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