Updated: Jul 12, 2026

Measurement of Fatty Acid β-Oxidation in a Suspension of Freshly Isolated Mouse Hepatocytes
Published on: September 9, 2021
Jean-Louis Gallis1, Pierre Tissier, Henri Gin
1Centre de Résonance Magnétique des Systèmes Biologiques, UMR 5536 CNRS-UB2, 146 rue Léo Saignat, 33076 F-Bordeaux Cedex, France. jean.louis.gallis@free.fr
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This study investigated how butyrate, a short-chain fatty acid, affects energy production in the liver of fed rats. Researchers measured ATP synthesis rates and oxygen consumption in livers perfused with butyrate, acetate, or octanoate. They found that butyrate significantly reduced ATP turnover and the ATP/O ratio compared to control conditions. Acetate had no significant effect, while octanoate also decreased ATP synthesis. The results suggest that butyrate alters liver metabolism similarly to longer-chain fatty acids. These findings highlight the need to consider the metabolic impact of butyrate when designing nutritional or therapeutic strategies involving short-chain fatty acids.
Area of Science:
Background:
The role of short-chain fatty acids (SCFAs) in hepatic metabolism remains partially understood. While SCFAs like butyrate are known to serve as energy sources for colonocytes, their impact on liver function is less clear. Prior research has shown that SCFAs may alter ATP levels and respiration in liver tissue. However, the specific effect of butyrate on oxidative phosphorylation in the liver has not been fully explored. This gap motivated the current investigation into how butyrate influences ATP synthesis and mitochondrial efficiency. It was already known that SCFAs can affect liver respiration, but the mechanism remains unclear. The study aimed to determine whether butyrate alters oxidative phosphorylation yield in fed rats. No prior work had resolved the exact impact of butyrate on ATP/O ratios in whole liver perfusion. This uncertainty drove the need for direct experimental analysis of hepatic metabolic responses to SCFAs.
Purpose Of The Study:
The study aimed to investigate how butyrate affects oxidative phosphorylation yield in liver tissue from fed rats. Specifically, the researchers sought to compare butyrate’s effects with those of acetate and octanoate. The primary objective was to determine the ATP/O ratio in whole liver perfusion under different SCFA conditions. The motivation stemmed from the need to understand how SCFAs influence hepatic energy production. By measuring ATP synthesis rates and oxygen consumption, the study aimed to clarify the metabolic impact of butyrate. The researchers also wanted to assess whether butyrate behaves similarly to longer-chain fatty acids in this context. The study’s design allowed for direct comparison of SCFA effects on liver bioenergetics. This work could help refine nutritional strategies involving SCFAs by identifying their metabolic consequences.
Butyrate significantly reduces ATP turnover and the ATP/O ratio in perfused liver tissue compared to control conditions.
ATP synthesis was estimated using 31P nuclear magnetic resonance and measured liver respiration rates after enzyme inhibition.
Octanoate was used as a longer-chain fatty acid to compare its effects on ATP/O ratio with those of butyrate and acetate.
31P NMR was used to estimate ATP flux and determine the ATP/O ratio in whole liver perfusion.
The ATP/O ratio in the control condition was 0.30 ± 0.05, measured in micromoles per minute per gram of liver tissue.
Main Methods:
The study used isolated, perfused livers from fed rats to examine the effects of SCFAs on oxidative phosphorylation. The researchers monitored ATP synthesis rates by measuring ATP loss after inhibiting cytochrome oxidase and glyceraldehyde 3-phosphate dehydrogenase. They employed 31P nuclear magnetic resonance to estimate ATP flux and measured liver respiration to calculate the ATP/O ratio. The experimental setup included perfusion with butyrate, acetate, and octanoate at 3 mmol/L concentrations. The ATP turnover was quantified in micromoles per minute per gram of liver tissue. Mitochondrial oxygen consumption was also recorded to assess metabolic activity. The study compared ATP/O ratios across different SCFA conditions to determine their impact on oxidative phosphorylation. This approach allowed the researchers to evaluate how each SCFA affects hepatic energy production.
Main Results:
The ATP turnover rate was significantly lower in the presence of butyrate (0.40 ± 0.10 micromoles/min.g) and octanoate (0.56 ± 0.10 micromoles/min.g) compared to control (1.09 ± 0.13 micromoles/min.g). Acetate perfusion did not cause a significant decrease (0.76 ± 0.10 micromoles/min.g). Mitochondrial oxygen consumption remained unchanged with acetate but increased significantly with butyrate and octanoate. The ATP/O ratio was notably reduced with butyrate (0.07 ± 0.02) and octanoate (0.09 ± 0.02) compared to control (0.30 ± 0.05). Acetate perfusion resulted in an ATP/O ratio of 0.20 ± 0.02, which was not significantly different from control. These findings suggest that butyrate and octanoate decrease ATP synthesis rates. The observed changes in ATP/O ratio indicate altered oxidative phosphorylation efficiency. These results support the hypothesis that butyrate affects hepatic metabolism similarly to longer-chain fatty acids.
Conclusions:
The authors propose that butyrate reduces the rate of ATP synthesis in perfused liver tissue from fed rats. This effect is reflected in a lower ATP/O ratio compared to control conditions. The findings suggest that butyrate behaves similarly to longer-chain fatty acids in terms of hepatic metabolic impact. The study does not claim that butyrate is essential for liver function but highlights its potential to alter energy production. The observed decrease in ATP turnover and ATP/O ratio indicates a shift in oxidative phosphorylation efficiency. The authors do not suggest that butyrate is harmful but emphasize the need to consider its metabolic effects in therapeutic or nutritional strategies. The results support the idea that SCFAs may influence liver metabolism when present in large quantities. These findings may inform future studies on the role of SCFAs in hepatic energy metabolism.
The authors suggest that butyrate alters hepatic energy production and should be considered in therapeutic or nutritional strategies involving SCFAs.