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  • RNA sequencing reveals niche gene expression effects of beta-hydroxybutyrate in primary myotubes.

RNA sequencing reveals niche gene expression effects of beta-hydroxybutyrate in primary myotubes.

Life science alliance (2021-08-20)
Philip Mm Ruppert, Lei Deng, Guido Jej Hooiveld, Roland Wj Hangelbroek, Anja Zeigerer, Sander Kersten
ABSTRACT

Various forms of fasting and ketogenic diet have shown promise in (pre-)clinical studies to normalize body weight, improve metabolic health, and protect against disease. Recent studies suggest that β-hydroxybutyrate (βOHB), a fasting-characteristic ketone body, potentially acts as a signaling molecule mediating its beneficial effects via histone deacetylase inhibition. Here, we have investigated whether βOHB, in comparison to the well-established histone deacetylase inhibitor butyrate, influences cellular differentiation and gene expression. In various cell lines and primary cell types, millimolar concentrations of βOHB did not alter differentiation in vitro, as determined by gene expression and histological assessment, whereas equimolar concentrations of butyrate consistently impaired differentiation. RNA sequencing revealed that unlike butyrate, βOHB minimally impacted gene expression in primary adipocytes, macrophages, and hepatocytes. However, in myocytes, βOHB up-regulated genes involved in the TCA cycle and oxidative phosphorylation, while down-regulating genes belonging to cytokine and chemokine signal transduction. Overall, our data do not support the notion that βOHB serves as a powerful signaling molecule regulating gene expression but suggest that βOHB may act as a niche signaling molecule in myocytes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(R)-(−)-3-Hydroxybutyric acid sodium salt, optical purity ee: 99% (GLC)
Sigma-Aldrich
Collagenase from Clostridium histolyticum, suitable for release of physiologically active rat epididymal adipocytes, Type II, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
Sigma-Aldrich
β-Hydroxybutyrate Assay Kit, sufficient for 100 colorimetric tests
Sigma-Aldrich
Sodium butyrate, 98%
Sigma-Aldrich
Dispase® II, protease