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. 2020 Aug 8;12(8):2379.
doi: 10.3390/nu12082379.

Mechanism of Action of Ketogenic Diet Treatment: Impact of Decanoic Acid and Beta-Hydroxybutyrate on Sirtuins and Energy Metabolism in Hippocampal Murine Neurons

Partha Dabke  1 , Anibh M Das  1
Affiliations

Mechanism of Action of Ketogenic Diet Treatment: Impact of Decanoic Acid and Beta-Hydroxybutyrate on Sirtuins and Energy Metabolism in Hippocampal Murine Neurons

Partha Dabke et al. Nutrients. .

Abstract

The ketogenic diet (KD), a high-lipid and low-carbohydrate diet, has been used in the treatment of epilepsy, neurodegenerative disorders, inborn errors of metabolism and cancer; however, the exact mechanism/s of its therapeutic effect is not completely known. We hypothesized that sirtuins (SIRT)-a group of seven NAD-dependent enzymes and important regulators of energy metabolism may be altered under KD treatment. HT22 hippocampal murine neurons were incubated with two important KD metabolites-beta-hydroxybutyrate (BHB) (the predominant ketone body) and decanoic acid (C10), both accumulating under KD. Enzyme activity, protein, and gene expressions of SIRT 1-4, enzyme capacities of the mitochondrial respiratory chain complexes (MRC), citrate synthase (CS) and gene expression of monocarboxylate transporters were measured in control (untreated) and KD-treated cells. Incubation with both-BHB and C10 resulted in significant elevation of SIRT1 enzyme activity and an overall upregulation of the MRC. C10 incubation showed prominent increases in maximal activities of complexes I + III and complex IV of the MRC and ratios of their activities to that of CS, pointing towards a more efficient functioning of the mitochondria in C10-treated cells.

Keywords: beta-hydroxybutyrate; decanoic acid; energy metabolism; ketogenic diet; mitochondria; sirtuins.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Graphs (A–C) represent absolute enzyme activities of SIRT 1–3 (Units/µg)., (D–F) represent relative protein expression, (G–I) are representative images of Western blots and (J–L) represent relative gene expression of SIRT 1–3. Data are expressed as mean + SD (n = 3–4).
Figure 1
Figure 1
Graphs (A–C) represent absolute enzyme activities of SIRT 1–3 (Units/µg)., (D–F) represent relative protein expression, (G–I) are representative images of Western blots and (J–L) represent relative gene expression of SIRT 1–3. Data are expressed as mean + SD (n = 3–4).
Figure 2
Figure 2
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity (Units/µg), (D–F,M) represent relative protein expression and (J–L) represent relative gene expression of SIRT 1–4. (G–I,N,O) are representative images of Western blots. * p < 0.05, ** p < 0.01; (n = 3–8). Data are represented as mean + SD.
Figure 2
Figure 2
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity (Units/µg), (D–F,M) represent relative protein expression and (J–L) represent relative gene expression of SIRT 1–4. (G–I,N,O) are representative images of Western blots. * p < 0.05, ** p < 0.01; (n = 3–8). Data are represented as mean + SD.
Figure 2
Figure 2
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity (Units/µg), (D–F,M) represent relative protein expression and (J–L) represent relative gene expression of SIRT 1–4. (G–I,N,O) are representative images of Western blots. * p < 0.05, ** p < 0.01; (n = 3–8). Data are represented as mean + SD.
Figure 3
Figure 3
Graphs (A–C,G,I) represent maximal enzymatic activities of the respiratory chain complexes I–IV, ATP–synthase (complex V) and citrate synthase. The graphs (D–F,H) depict normalized ratios of the enzymatic activities of the respiratory chain to the enzymatic activity of citrate synthase–the mitochondrial marker enzyme. * p < 0.05 (n = 3–4). Data are shown as mean + SD.
Figure 3
Figure 3
Graphs (A–C,G,I) represent maximal enzymatic activities of the respiratory chain complexes I–IV, ATP–synthase (complex V) and citrate synthase. The graphs (D–F,H) depict normalized ratios of the enzymatic activities of the respiratory chain to the enzymatic activity of citrate synthase–the mitochondrial marker enzyme. * p < 0.05 (n = 3–4). Data are shown as mean + SD.
Figure 4
Figure 4
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity., (D–F,M) represent relative protein expression and (J–L,N) represent relative gene expression of SIRT 1–4. (G–I,O) are representative western blot images. * p < 0.05 (n = 3–6). Data are represented as mean + SD.
Figure 4
Figure 4
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity., (D–F,M) represent relative protein expression and (J–L,N) represent relative gene expression of SIRT 1–4. (G–I,O) are representative western blot images. * p < 0.05 (n = 3–6). Data are represented as mean + SD.
Figure 4
Figure 4
Graphs (A–C) represent absolute SIRT 1–3 enzyme activity., (D–F,M) represent relative protein expression and (J–L,N) represent relative gene expression of SIRT 1–4. (G–I,O) are representative western blot images. * p < 0.05 (n = 3–6). Data are represented as mean + SD.
Figure 5
Figure 5
Graphs (A–C,G,I) represent maximal enzymatic activities of the respiratory chain complexes I–IV, ATP–synthase (complex V) and citrate synthase (CS). The graphs (D–F,H) depict ratios of the enzymatic activities of the respiratory chain to the enzymatic activity of citrate synthase–the mitochondrial marker enzyme. * p < 0.05; ** p < 0.01 (n = 3–4). Data are represented as mean + SD.
Figure 5
Figure 5
Graphs (A–C,G,I) represent maximal enzymatic activities of the respiratory chain complexes I–IV, ATP–synthase (complex V) and citrate synthase (CS). The graphs (D–F,H) depict ratios of the enzymatic activities of the respiratory chain to the enzymatic activity of citrate synthase–the mitochondrial marker enzyme. * p < 0.05; ** p < 0.01 (n = 3–4). Data are represented as mean + SD.
Figure 6
Figure 6
Gene expression of monocarboxylate transporters (MCTr 1 and MCTr 2) in 5 mM ßHB (A,B) or 250 µM C10-treated HT22 cells (C,D), relative to control (untreated) probes. Data are represented as mean + SD; * p < 0.05 (n = 3).
Figure 7
Figure 7
Schematic representation of a neuronal cell with sirtuins (Sirtuins 1–4) and respiratory chain and effects of beta-hydroxybutyrate (ßHB) on the same. Purple dotted arrows represent the known regulatory effects of SIRT 1–4. Solid red and blue arrows represent the effects of 5 mM ßHB and 250 µM C10, respectively. The light-yellow circle represents the nucleus with DNA and the light purple triangle therein represents transcription factors deacetylated by SIRT1 (PGC-1α and FOXO3a). ßHB—beta hydroxybutyrate; C10—decanoic acid; CS—citrate synthase; IMM—inner mitochondrial membrane; OMM—outer mitochondrial membrane; MRC—mitochondrial respiratory chain; ROS—reactive oxygen species; SIRT—sirtuin.
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References

    1. Wheless J.W. History of the ketogenic diet. Epilepsia. 2008;49:3–5. doi: 10.1111/j.1528-1167.2008.01821.x. - DOI - PubMed
    1. Huttenlocher P.R. Ketonemia and Seizures: Metabolic and Anticonvulsant Effects of Two Ketogenic Diets in Childhood Epilepsy. Pediatr. Res. 1976;10:536–540. doi: 10.1203/00006450-197605000-00006. - DOI - PubMed
    1. Huttenlocher P.R., Wilbourn A.J., Signore J.M. Medium-chain triglycerides as a therapy for intractable childhood epilepsy. Neurology. 1971;21:1097–1103. doi: 10.1212/WNL.21.11.1097. - DOI - PubMed
    1. Hartman A.L., Vining E.P. Clinical aspects of the ketogenic diet. Epilepsia. 2007;48:31–42. doi: 10.1111/j.1528-1167.2007.00914.x. - DOI - PubMed
    1. Gano L.B., Patel M., Rho J.M. Ketogenic diets, mitochondria, and neurological diseases. J. Lipid Res. 2014;55:2211–2228. doi: 10.1194/jlr.R048975. - DOI - PMC - PubMed

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