Netrin-1 upregulates the its own gene expression and unc5b gene expression, but downregulates ccl19 gene expression in human macrophages THP-1 cell line

According to the WHO data of 2021, ischemic heart disease was the leading cause of human disability and mortality at a global level, with brain stroke being the third leading cause of death. These cardiovascular diseases, including myocardial infarction and aortic aneurysms, are largely caused by atherosclerosis, a systemic chronic inflammatory process. This disease develops over decades and is characterized by the disruption of the vascular endothelial layer integrity, excessive lipid accumulation in arterial walls, macrophage and smooth muscle cells dysfunction, and evolving inflammation of arterial walls. One of approaches to atherosclerosis regression may be emigration of cholesterol ester-laden macrophage from atherosclerotic plaque via lymphatic vessels to the regional lymph nodes, and then to liver for subsequent cholesterol metabolism. The protein netrin-1 is inhibitor of immune cells migration. Particular interest is the study of the autocrine/paracrine regulation of netrin-1 gene expression in human macrophages. The understanding of this process may provide a basis for discovery of the new approaches to atherosclerosis therapy. The aim of the present article was to study the influence of protein factor netrin-1 on the gene expression levels of ntn1, its receptor unc5b, and ccl19. The experiments were carried out on the macrophage-like cells derived from acute monocytic leukemia cell line THP-1. The expression of ntn1, unc5b, ccl19 genes was performed by reverse transcription and real-time quantitative PCR (mRNA level), and by flow cytometry (at the protein level). Netrin-1 at the concentration of 50 ng/mL induced ntn1 and unc5b gene expression, but suppressed ccl19 gene expression in human macrophages. However, netrin-1 applied at the concentration of 250 ng/mL caused a significant downregulation of the own ntn1 gene expression. Netrin-1 has autoregulatory effect on its own gene expression, and on the gene encoding its receptor (unc5b) in human macrophage-like cells. The downregulation of chemokine ccl19 expression in macrophages mediated by netrin-1 may be a potential way by which this effector inhibits macrophage emigration. The obtained results suggest a regulatory mechanism for expression of proteins responsible for macrophage migration, which in the future will enable application of these data for searching new targets in atherosclerosis therapy.

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