Inhibition of anti-inflammatory cytokines by the drug “Alvostaz” in the in vitro system

The aim of the work is to study the molecular mechanism of action of the drug “Alvostaz”.

The following scheme was used to evaluate the molecular mechanism of action of “Alvostaz”: to trigger inflammation and cytokine synthesis, cells were stimulated by the inflammatory trigger TNF-alpha (1 series of experiments), and to assess the biological activity of “Alvostaz”, cells were treated with a mixture of TNF-α and “Alvostaz” in different concentrations (2 series of experiments). The mediated effect of TNF-α on cells was assessed by the level of secretion of cytokines IL-6 and MCP-1, involved in the pathogenesis of alveolar inflammation and the maintenance of bone homeostasis. When inflammation was stimulated by TNF, fibroblasts dramatically increased the production of MCP-1 and IL-6. In the first series of experiments, optimal concentrations of the inflammatory trigger TNF-α and “Alvostaz” were selected, at which human fibroblasts remained viable and responded to inflammation stimulation by synthesizing cytokines IL-6 and MCP-1.  An increase in MCP-1 concentrations in the culture medium correlated with a decrease in the concentration of “Alvostaz”, which indicates the MCP-1 inhibitory effect of “Alvostaz”. Under the same conditions, the production of IL-6 by fibroblasts is exactly the opposite in nature compared to MCP-1. By itself, “Alvostaz” does not cause synthesis and secretion of cytokine IL-6 into the extracellular medium by unstimulated fibroblasts without the addition of TNF. TNF-stimulated fibroblasts demonstrated a decrease in IL-6 production in proportion to a decrease in the dose of “Alvostaz” in the culture medium, which indicates a correlation between IL-6 synthesis and the presence of “Alvostaz” in the cell culture medium.

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