NETRIN-1 UPREGULATES THE ITS OWN GENE EXPRESSION AND UNC5B GENE EXPRESSION, BUT DOWNREGULATES CCL-19 GENE EXPRESSION IN HUMAN MACROPHAGES THP-1 CELL LINE

Abstract

The relevance of the theme: According to the World Health Organization data on 2021 the leading cause of human disability and mortality was ischemic heart disease at a global level, stroke was the third leading cause of death. These cardiovascular diseases, including myocardial infarction and aortic aneurysms, are caused by a systemic chronic inflammatory process called atherosclerosis. This disease develops over decades and is characterized by the disruption of the vascular endothelial layer integrity, the excessive lipid accumulation in the arterial walls, macrophage and smooth muscle cells dysfunction and the inflammation of the arterial walls development. One of the ways for atherosclerosis regression may be cholesterol ester-laden macrophage emigration from atherosclerotic plaque through the lymphatic vessels to the regional lymph nodes and then to the liver for the subsequent cholesterol metabolism. The protein netrin-1 is the 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 be basis for a discovery of the new approaches in atherosclerosis therapy.

The aim of the article: to study the influence of netrin-1 protein on the expression level NTN1 gene, its receptor UNC5B gene and CCL19 gene.

Materials and methods: The experiments were carried out on the macrophages differentiated from acute monocytic leukemia cell line THP-1. The analysis of NTN1, UNC5B, CCL19 genes expression was performed by methods the reverse transcription and real time quantitative polymerase chain reaction (on the messenger RNA level) and by flow cytometry (on the protein level).

Results: Netrin-1 in the concentration of 50 ng/ml induced NTN1 and UNC5B genes expression, but suppressed CCL19 gene expression in human macrophages. However, netrin-1 in the concentration of 250 ng/ml significant downregulated of its own  NTN1 gene expression

Conclusions: Netrin-1 has autoregulatory effect on the expression of its own gene and the gene of its receptor UNC5B in human macrophages. Netrin-1 mediated downregulation of chemokine CCL19 expression in macrophages may be one of the ways which this effector inhibits macrophage emigration. The obtained results expand the insight of the regulation of the expression of proteins responsible for macrophage migration, which in the future will allow performing these data to research new targets in atherosclerosis therapy.

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