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Technical Briefs

Differential Translocation of Nuclear Factor-KappaB in a Cardiac Muscle Cell Line Under Gravitational Changes

[+] Author and Article Information
Ohwon Kwon

Department of Mechanical Engineering, University of Cincinnati, Cincinnati, OH 45221

Michael Tranter, W. Keith Jones

Department of Pharmacology and Cell Biophysics, University of Cincinnati, Cincinnati, OH 45221

John M. Sankovic

Microgravity Science Division, NASA Glenn Research Center, Cleveland, OH 44135

Rupak K. Banerjee1

Department of Mechanical Engineering, and Department of Biomedical Engineering, University of Cincinnati, Cincinnati, OH 45221rupak.banerjee@uc.edu

1

Corresponding author.

J Biomech Eng 131(6), 064503 (May 08, 2009) (4 pages) doi:10.1115/1.3128718 History: Received February 08, 2009; Revised March 20, 2009; Published May 08, 2009

Abstract

Microgravity (micro-g) environments have been shown to elicit dysregulation of specific genes in a wide assay of cell types. It is known that the activation of transcription factors and molecular signaling pathways influence various physiological outcomes associated with stress and adaptive responses. Nuclear factor-kappa B $(NF-κB)$ is one of the most prevailing oxidation-sensitive transcription factors. It is hypothesized that simulated microgravity would activate $NF-κB$ and its downstream transcriptional networks, thus suggesting a role for $NF-κB$ in microgravity induced muscle atrophy. To investigate the activation of $NF-κB$ in a rat cardiac cell line (H9c2) under micro-g, rotating wall vessel bioreactors were used to simulate micro-g conditions. Western blotting revealed that mean nuclear translocation of $NF-κB$ p65 subunit was 69% for micro-g and 46% for unit-g dynamic control as compared with a 30 min $TNF-α$ positive control $(p<0.05, n=3)$. The results from western blots were confirmed by enzyme-linked immunosorbent assay, which showed 66% for micro-g and 45% for dynamic control as compared with positive control $(p<0.05, n=3)$. These results show significant differential translocation of $NF-κB$ p65 under simulated micro-g. These results may be expanded upon to explain physiological changes such as muscle atrophy and further identify the regulatory pathways and effector molecules activated under exposure to micro-g.

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Figures

Figure 1

Experimental design: (a) RWVs in the simulated MG and (b) the unit-g DC

Figure 2

Western blots result for NF-κB p65 in nuclear lysates of H9c2 cells (p∗<0.05, data are expressed as mean ± SEM of three independent experiments)

Figure 3

ELISA result for NF-κB p65 in nuclear lysates of H9c2 cells (p∗<0.05, data are expressed as mean ± SEM of three independent experiments)

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