Single-cell RNA sequencing analyses the mechanism of scar-associated macrophages promoting hepatic stellate cell activation
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.
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NSTL
万方数据
维普
