In vitro γ-ray-induced inflammatory response is dominated by culturing conditions rather than radiation exposures

Scientific Reports
G BabiniA Ottolenghi

Abstract

The inflammatory pathway has a pivotal role in regulating the fate and functions of cells after a wide range of stimuli, including ionizing radiation. However, the molecular mechanisms governing such responses have not been completely elucidated yet. In particular, the complex activation dynamics of the Nuclear transcription Factor kB (NF-kB), the key molecule governing the inflammatory pathway, still lacks a complete characterization. In this work we focused on the activation dynamics of the NF-kB (subunit p65) pathway following different stimuli. Quantitative measurements of NF-kB were performed and results interpreted within a systems theory approach, based on the negative feedback loop feature of this pathway. Time-series data of nuclear NF-kB concentration showed no evidence of γ-ray induced activation of the pathway for doses up to 5 Gy but highlighted important transient effects of common environmental stress (e.g. CO2, temperature) and laboratory procedures, e.g. replacing the culture medium, which dominate the in vitro inflammatory response.

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Methods Mentioned

BETA
environmental stresses
Protein Assay
Assay
Enzyme-Linked ImmunoSorbent Assay
ELISA
environmental stress
nuclear translocation

Software Mentioned

MRI
Gnuplot
Cell Image Analyzer
MINUIT
ImageJ

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