Effect of ketogenic diet and beta-hydroxybutyrate on NF-kappaB activity
Inflammation is the driver of many modern lifestyle diseases including cancer, autoimmune diseases, diabetes and cardiovascular disease. Results from a number of studies have emphasized the importance of lifestyle and nutrition in preventing or reducing low-grade inflammation. Certain low-carbohydrate diets such as the ketogenic diet have gained popularity over the recent years. During ketogenic diet, fasting and strenous exercise, ketobodies such as b-hydroxybutyrate (BHB) can accumulate in the blood and recent studies have demonstrated that BHB can act as an anti-inflammatory molecule. Specifically it has been shown to inhibit components of the inflammasome and thereby reduce secretion of potent proinflammatory cytokines including IL1-beta. We therefore hypothesize that interventions with the aim of increasing BHB can mediate an anti-inflammatory effect, and we plan here to perform experiments to elucidate this.
We suggest that BHB affects the immune system by directly and indirectly downregulating several pathways integral to the immune system. A key regulator of inflammation is the transcription factor NF-kB. We have generated transgenic reporter mice that contain binding sites for NF-kB coupled to the reporter gene luciferase. Luciferase activity is therefore increased during inflammation and can be visualised as increased light production inside the mouse. By using these transgenic reporter mice we can observe and quantify changes of the inflammatory response using a light sensitive camera (IVIS). In this experiment we plan two approaches to test the impact of BHB on inflammation: 1) Feed NF-kB luciferase with a ketogenic diet with or without a short fasting period or 2) Administer known concentrations of BHB systemically in the mouse. In both cases we will challenge mice with a low but profinflammatory dose of lipopolysaccharide (LPS) from E.coli and measure whether BHB and or ketogenic diet +/- fasting exerts an antiinflammatory effect. This experiment will shed light on potential health benefits of raising systemic BHB levels.
Number of animals applied for is 102.
These experiments cannot be easily modelled or mimicked by in vitro studies. The immune system is a complex system of many important pathways, therefore, to achieve representative data that would be more akin to a human response, a mouse model is preferred. The mice will be monitored daily, looking for any visual signs of discomfort, and weighed during the diet portion of the experiment. Since LPS will be used, the use of a pain reliever (e.g. Temgesic) is considered if the mice show signs of significant discomfort. However, the LPS doses used will be relatively low, so from experience we do not expect any great discomfort for the mice.
We suggest that BHB affects the immune system by directly and indirectly downregulating several pathways integral to the immune system. A key regulator of inflammation is the transcription factor NF-kB. We have generated transgenic reporter mice that contain binding sites for NF-kB coupled to the reporter gene luciferase. Luciferase activity is therefore increased during inflammation and can be visualised as increased light production inside the mouse. By using these transgenic reporter mice we can observe and quantify changes of the inflammatory response using a light sensitive camera (IVIS). In this experiment we plan two approaches to test the impact of BHB on inflammation: 1) Feed NF-kB luciferase with a ketogenic diet with or without a short fasting period or 2) Administer known concentrations of BHB systemically in the mouse. In both cases we will challenge mice with a low but profinflammatory dose of lipopolysaccharide (LPS) from E.coli and measure whether BHB and or ketogenic diet +/- fasting exerts an antiinflammatory effect. This experiment will shed light on potential health benefits of raising systemic BHB levels.
Number of animals applied for is 102.
These experiments cannot be easily modelled or mimicked by in vitro studies. The immune system is a complex system of many important pathways, therefore, to achieve representative data that would be more akin to a human response, a mouse model is preferred. The mice will be monitored daily, looking for any visual signs of discomfort, and weighed during the diet portion of the experiment. Since LPS will be used, the use of a pain reliever (e.g. Temgesic) is considered if the mice show signs of significant discomfort. However, the LPS doses used will be relatively low, so from experience we do not expect any great discomfort for the mice.