Profile of IL-6 and TNF in Foam Cell Formation: An Improved Method Using Fluorescein Isothiocyanate (FITC) Probe

Background: The formation of foam cells occurs due to the increase in low-density plasma lipoprotein (LDL) and dysregulation of inflammation, which is important for the development of atherosclerosis. Objective: To evaluate the profile of tumor necrosis factor-alpha (TNF-alpha) and Interleukin-6 (IL-6) in the existing foam cell formation method, optimizing this protocol. Methods: The LDL was isolated, oxidized, and labeled with a Fluorescein isothiocyanate (FITC) probe. Foam cells were generated from THP-1 human monocyte-derived cells and incubated in the absence (control) or presence of FITC-ox-LDL (10, 50, 100, 150, or 200 mg/mL), for 12, 24, 48, or 72 hours. The accumulated FITC-ox-LDL in the cell was quantified by microscopy. The enzyme-linked immunosorbent assay was evaluated to quantify the IL-6 and TNF-alpha, with p < 0.05 considered significant. Results: All the FITC-ox-LDL concentrations tested showed a higher fluorescence when compared to the control, showing a greater accumulation of lipoprotein in cells. The higher the concentration of FITC-ox-LDL, the greater the production of TNF-alpha and IL-6. The production of IL-6 by foam cells was detected up to the value of 150 mg/mL of the maximum stimulus for LDL. Concentrations above 50 mg/mL LDL stimulated greater release of TNF-alpha compared to control. Conclusions: Our model contributes to the understanding of the release of IL-6 and TNF-alpha in response to different concentrations of ox-LDL, using an optimized method for the formation of foam cells.

Automated summary

The study evaluated the role of TNF-α and IL-6 in foam cell formation, confirming that higher concentrations of FITC-ox-LDL led to more accumulation of lipoproteins in cells and higher production of TNF-α and IL-6.