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Exogenous H2S switches cardiac energy substrate metabolism by regulating SIRT3 expression in db/db mice

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Abstract

Hydrogen sulfide (H2S) is involved in diverse physiological functions, such as anti-hypertension, anti-proliferation, regulating ATP synthesis, and reactive oxygen species production. Sirtuin 3 (SIRT3) is a NAD+-dependent deacetylase that regulates mitochondrial energy metabolism. The role of H2S in energy metabolism in diabetic cardiomyopathy (DCM) may be related to regulate SIRT3 expression; however, this role remains to be elucidated. We hypothesized that exogenous H2S could switch cardiac energy metabolic substrate preference by lysine acetylation through promoting the expression of SIRT3 in cardiac tissue of db/db mice. Db/db mice, neonatal rat cardiomyocytes, and H9c2 cell line with the treatment of high glucose, oleate, and palmitate were used as animal and cellular models of type 2 diabetes. Using LC-MS/MS, we identified 76 proteins that increased acetylation, including 8 enzymes related to fatty acid β-oxidation and 7 enzymes of the tricarboxylic acid (TCA) cycle in the db/db mice hearts compared to those with the treatment of NaHS. Exogenous H2S restored the expression of NAMPT and the ratio of NAD+/NADH enhanced the expression and activity of SIRT3. As a result of activation of SIRT3, the acetylation level and activity of fatty acid β-oxidation enzyme LCAD and the acetylation of glucose oxidation enzymes PDH, IDH2, and CS were reduced which resulted in activation of PDH, IDH2, and CS. Our finding suggested that H2S induced a switch in cardiac energy substrate utilization from fatty acid β-oxidation to glucose oxidation in DCM through regulating SIRT3 pathway.

Key messages

  • H2S regulated the acetylation level and activities of enzymes in fatty acid oxidation and glucose oxidation in cardiac tissues of db/db mice.

  • Exogenous H2S decreased mitochondrial acetylation level through upregulating the expression and activity of SIRT3 in vivo and in vitro.

  • H2S induced a switch in cardiac energy substrate utilization from fatty acid oxidation to glucose.

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Abbreviations

C-7Az:

7-Azido-4-methylcoumarin

CPT1:

Carnitine palmitoyltransferase 1

CS:

Citrate synthase

CSE:

Cystathionine gamma-lyase

DCM:

Diabetic cardiomyopathy

ESM:

Electronic supplementary material

FAO:

Fatty acid β-oxidation

FFA:

Free fatty acid

GO:

Gene ontology

H2S:

Hydrogen sulfide

IDH:

Isocitrate dehydrogenase

KEGG:

Kyoto Encyclopedia of Genes and Genomes

LCAD:

Long-chain acyl-CoA dehydrogenase

Lys:

Lysine

NAD:

Nicotinamide adenine dinucleotides

NAM:

Nicotinamide

NAMPT:

Nicotinamide phosphoribosyltransferase

NMN:

Nicotinamide mononucleotide

PDH:

Pyruvate dehydrogenase

PPG:

DL-propargylglycine

RCR:

Respiratory control rate

SIRT:

Silent mating type information regulation 2 homolog

SIRT 3:

Sirtuin 3

TCA:

Tricarboxylic acid cycle

T2DM:

Type 2 diabetes mellitus

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Acknowledgements

We thank Jingjie PTM BioLab Co.Ltd. (Hangzhou, China) for the mass spectrometry analysis.

Contribution statement

Zhang Weihua, Lu Fanghao and Sun Yu designed, developed, and performed the majority of the experiments. Zhiliang Tian, Lin Ning, Linxue Zhang, Shiyun Dong and Zhao Yajun performed mitochondrial enzyme activities analysis. Ren Huan, He Chen, Fan Yang, Jichao Wu, Yan Wang and Dechao Zhao provided additional bioinformatic data analysis. Gao Zhaopeng, Xiaojiao Sun, Miao Yu and Changqing Xu performed immunoprecipitation analysis. Zhang Weihua and Lu Fanghao supervised and managed the project and edited large sections of the manuscript. All authors contributed to writing and revising the manuscript.

To whom correspondence should be addressed Department of Pathophysiology, Harbin Medical university, Harbin, Heilongjiang Province 150086. Tel.:451-86674548; E-mail: zhangwh116@126.com; lufanghao1973@126.com. These authors contributed equally to this work.

Funding

This study was supported by the National Natural Science Foundation of China (81670344, 81370421, 81370330) and the Natural Science Foundation of Heilongjiang (No. D201070).

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Authors and Affiliations

  1. Department of Pathophysiology, Harbin Medical University, Harbin, 150086, China

    Yu Sun, Ning Liu, Linxue Zhang, Zhaopeng Gao, Xiaojiao Sun, Miao Yu, Jichao Wu, Fan Yang, Yajun Zhao, Shiyun Dong, Changqing Xu, Fanghao Lu & Weihua Zhang

  2. Department of Pediatric, Second Clinical Medical School of Harbin Medical University, Harbin, 150001, China

    Zhiliang Tian

  3. Department of Immunology, Harbin Medical University, Harbin, 150086, China

    Huan Ren

  4. Department of Pathology, Harbin Medical University, Harbin, 150086, China

    He Chen

  5. Department of Cardiology, First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China

    Dechao Zhao

  6. Department of Urologic Surgery, First Clinical Medical School of Harbin Medical University, Harbin, 150001, China

    Yan Wang

  7. Key Laboratory of Cardiovascular Medicine Research, Harbin Medical University Ministry of Education, Harbin, 150086, China

    Weihua Zhang

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  1. Yu Sun
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Correspondence to Fanghao Lu or Weihua Zhang.

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Sun, Y., Tian, Z., Liu, N. et al. Exogenous H2S switches cardiac energy substrate metabolism by regulating SIRT3 expression in db/db mice. J Mol Med 96, 281–299 (2018). https://doi.org/10.1007/s00109-017-1616-3

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