Differences in the Effects of Beta-Hydroxybutyrate on Mitochondria Biogenesis, Markers of Oxidative Stress and Inflammation in Young and Old Rat Tissues

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Abstract

One of the therapeutic approaches to age-related diseases is to affect the metabolism of the body’s cells through certain diets or their pharmacological mimetics. The ketogenic diet significantly affects the energy metabolism of cells and the functioning of mitochondria, which is being actively studied in various age-related pathologies. In this study, we investigated the effect of the ketogenic diet mimetic beta-hydroxybutyrate (BHB) on the gene expression of proteins regulating mitochondrial biogenesis (Ppargc1a, Nrf1, Tfam), quality control (Sqstm1), the work of the antioxidant system (Nfe2l2, Gpx1, Gpx3, Srxn1, Txnrd2, Slc6a9, Slc7a11), and the inflammatory response (Il1b, Tnf, Ptgs2, Gfap) in the brain, lungs, heart, liver, kidneys, and muscles of young and old rats. In addition, we analyzed mitochondrial DNA (mtDNA) copy number, the accumulation of mtDNA damage, and the level of oxidative stress by the concentration of thiobarbituric acid-reactive substances (TBARS), and reduced glutathione level. We showed that aging in a number of organs disrupts mitochondrial biogenesis and the functioning of the cell’s antioxidant system, which was accompanied by increased oxidative stress and inflammation. Administration of BHB for 2 weeks had different effects on organs of young and old rats. In particular, BHB increased the expression of genes of proteins associated with mitochondrial biogenesis and the antioxidant system, especially in the liver tissue and muscles of the young but not the old rats. At the same time, BHB contributed to the reduction of TBARS in the kidneys of the old rats. Thus, our study has shown that the administration of ketone bodies can significantly affect gene expression in organs, especially in young rats, by increasing mitochondrial biogenesis, improving the antioxidant system and partially reducing the level of oxidative stress. However, these changes were much less pronounced in old animals.

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About the authors

V. V. Nesterova

Voronezh State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Voronezh

P. I. Babenkova

Voronezh State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Voronezh

A. A. Brezgunova

Lomonosov Moscow State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Moscow

N. A. Samoylova

Voronezh State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Voronezh

I. S. Sadovnikova

Voronezh State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Voronezh

D. S. Semenovich

Lomonosov Moscow State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Moscow

N. V. Andrianova

Lomonosov Moscow State University

Email: plotnikov@belozersky.msu.ru
Russian Federation, Moscow

A. P. Gureev

Voronezh State University; Voronezh State University of Engineering Technology

Email: plotnikov@belozersky.msu.ru
Russian Federation, Voronezh; Voronezh

E. Y. Plotnikov

Lomonosov Moscow State University

Author for correspondence.
Email: plotnikov@belozersky.msu.ru
Russian Federation, Moscow

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2. Fig. 1. Effect of aging on gene expression of proteins responsible for mitochondrial biogenesis (a-c), autophagy (d), cellular antioxidant system (e-m) and inflammation (n-r). Results are presented as mean values ± error of the mean. Reliability of differences between groups: * p < 0.05 (t-test)

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3. Fig. 2. Effect of subcutaneous administration of BGB for 2 weeks on tissues of young rats. Effect of BGB on the expression of proteins responsible for mitochondrial biogenesis and antioxidant system in liver (a-z) and muscle (i-m) tissues. Results are presented as mean values ± error of the mean. Reliability of differences between groups: * p < 0.05 (t-criterion)

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4. Fig. 3. Heat maps showing the effects of age and subcutaneous injections of BHB on gene expression levels in rat brain, lung, heart, liver, kidney and muscle

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5. Fig. 4. Effect of subcutaneous injections of BHB on the number of mtDNA copies in different tissues of rats of different ages. yDNA - nuclear DNA. Results are presented as mean values ± error of the mean. Reliability of differences between groups: * p < 0.05; ** p < 0.01; *** p < 0.001 (Kraskell-Wallis test)

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6. Fig. 5. Effect of subcutaneous injections of BHB on the amount of mtDNA lesions in different tissues of rats of different ages. Results are presented as mean values ± error of the mean. Significance of differences with the control group of 4-month-old rats: * p < 0.05; ** p < 0.01 (Kraskell-Wallis test)

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7. Fig. 6. Effect of subcutaneous injections of BHB on the number of large-scale mtDNA deletions in different tissues of rats of different ages. Results are presented as mean values ± error of the mean. The significance of differences between groups: * p < 0.05; ** p < 0.01 (Kraskell-Wallis test)

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8. Fig. 7. Effect of subcutaneous injections of BHB on TBCRS content in different tissues of rats of different ages. The results are presented as mean values ± error of the mean. The significance of differences between groups: * p < 0.05; ** p < 0.01; *** p < 0.001 (Kraskell-Wallis test)

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9. Fig. 8. Effect of subcutaneous injections of BHB on the content of reduced glutathione in different tissues of young and old rats. Results are presented as mean values ± error of the mean. Significance of differences between groups: *** p < 0.001 (Kraskell-Wallis test)

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