Introduction: Treatment with bacterial lipopolysaccharides (LPS) is a convenient model used for stimulation of inflammatory response in cell cultures, which is also suggested to be associated with provoked antioxidant defense. Well defined markers are needed to verify pronounced cellular response in this model.
Aim: The aim of current study was to measure changes in expression levels of selected genes in order to identify indicative markers for verification of induced cellular response in a model of LPS-treated J744A.1 macrophages.
Materials and Methods: In order to determine most appropriate LPS treatment concentration, an MTT test was performed. LPS was applied in different concentrations (50 - 300 ng/mL) and their effect on the cell viability of J744A.1 macrophages was measured.
Results and Discussion: Analysis of the results of the MTT test showed a statistically significant (p <0.001) and equal effect of the three LPS concentrations (100, 200 and 300 ng/mL) applied. Cell viability was decreased with 20%. The effect of 100 ng/mL LPS treatment on the following genes was evaluated: antioxidant defense-related (GCLc, GPx1, GSS, GR and SOD2); inflammation-related (IL1β, IL6, MCP1, TNFα, IL1RN, NOS2, CRP, COX2); phagocytosis-related (NOX1 and MPO), and LPS/TLR4 signaling cascade-related (TLR4, IKK2, NRF1, NQO1). All of the studied genes were significantly induced upon LPS treatment for 24h determining it is sufficient to provoke pronounced cellular response. However, the following genes appear to be most affected by 24h LPS treatment: GCLc, COX2, NOS2, IL6, IL1β, CRP, NOX, TLR4 and IKK2.
Conclusion: In conclusion, studied genes may serve as suitable indicative markers for triggered cellular response were LPS stimulation of J774A.1 cells is about to be used as a model of oxidative and inflammatory provocation.
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