Exosome‐derived miR‐142‐5p from liver stem cells improves the progression of liver fibrosis by regulating macrophage polarization through CTSB

Abstract

Background This study aims to explore the effect of liver stem cells (LSCs)‐derived exosomes and the miR‐142a‐5p carried by them on the process of fibrosis by regulating macrophages polarization. Methods In this study, CCL4 was used to establish liver fibrosis model. The morphology and purity of exosomes (EVs) were verified by transmission electron microscopy, western blotting (WB) and nanoparticle tracing analysis (NTA). Real‐time quantitative PCR (qRT‐PCR), WB and enzyme‐linked immunoadsorption (ELISA) were used to detect liver fibrosis markers, macrophage polarization markers and liver injury markers. Histopathological assays were used to verify the liver injury morphology in different groups. The cell co‐culture model and liver fibrosis model were constructed to verify the expression of miR‐142a‐5p and ctsb. Results Immunofluorescence of LSCs markers CK‐18, epithelial cell adhesion molecule (EpCam), and AFP showed that these markers were up‐regulated in LSCs. In addition, we evaluated the ability of LSCs to excrete EVs by labeling LSCs‐EVs with PKH67. We found that CCL4 and EVs were simultaneously treated at 50 and 100 μg doses, and both doses of EVs could reduce the degree of liver fibrosis in mice. We tested markers of M1 or M2 macrophage polarization and found that EVs reduced M1 marker expression and promoted M2 marker expression. Further, ELISA was used to detect the secreted factors related to M1 and M2 in tissue lysates, which also verified the above views. Further analysis showed that the expression of miR‐142a‐5p increased significantly with the increase of EVs treatment concentration and time. Further, in vitro and in vivo LSCs‐EVs regulate macrophage polarization through miR‐142a‐5p/ctsb pathway and affect the process of liver fibrosis. Conclusion Our data suggest that EVs‐derived miR‐142‐5p from LSCs improves the progression of liver fibrosis by regulating macrophage polarization through ctsb.

Extracellular vesicles targeting non-parenchymal cells: the therapeutical effect on liver fibrosis

“…This results in decreased markers of M1 macrophage polarisation and increased markers of M2 macrophage polarisation. 75 Gingival tissue-derived MSCs (GMSCs) are Open access the mesenchymal stem cells of dental origin, with unique immunoregulatory capacity and secrete large amounts of EVs. 76 In a recent study, Watanabe et al found that EVs derived from GMSCs primed with a combination of two proinflammatory cytokines, TNF-α and interferon-α, synergistically promote anti-inflammatory M2 macrophage polarisation by increasing the expression of cluster of differentiation 73 (CD73) and CD5 molecule-like in EVs, which are mediated by mammalian target of rapamycin hypoxia-inducible factor-1α axis.…”

Role of Exosomal Modulation of Macrophages in Liver Fibrosis

“… 101 In another study, scientists examined changes in exosomes produced by liver stem cells in a CCl 4 -induced fibrosis model, and they identified the increased expression of miR-142a-5p, which prevents macrophages toward M1 polarization and facilitated the production of M2 markers through targeting cathepsin B to suppress LF in mice. 102 …”

“…101 In another study, scientists examined changes in exosomes produced by liver stem cells in a CCl 4 -induced fibrosis model, and they identified the increased expression of miR-142a-5p, which prevents macrophages toward M1 polarization and facilitated the production of M2 markers through targeting cathepsin B to suppress LF in mice. 102 Artificial exosome system: Artificial exosomes have been explored to treat LF in recent decades. Contemporary scholarly investigations are mainly concentrated on utilizing unaltered or modified exosomes as an innovative approach for drug administration, explicitly targeting the liver to deliver antifibrotic medications with precision.…”

“…116 Generally speaking, in the progression of LF, macrophages are the core hub of the regulation network (Table 2). 98,[100][101][102][103][104][105][106]109,110,112,117…”