Exogenous H2S switches cardiac energy substrate metabolism by regulating SIRT3 expression in db/db mice
- 406 Downloads
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.
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.
KeywordsHydrogen sulfide (H2S) Surtuin 3 Acetylation Fatty acid β-oxidation Glucose oxidation
Carnitine palmitoyltransferase 1
Electronic supplementary material
Fatty acid β-oxidation
Free fatty acid
Kyoto Encyclopedia of Genes and Genomes
Long-chain acyl-CoA dehydrogenase
Nicotinamide adenine dinucleotides
Respiratory control rate
Silent mating type information regulation 2 homolog
- SIRT 3
Tricarboxylic acid cycle
Type 2 diabetes mellitus
We thank Jingjie PTM BioLab Co.Ltd. (Hangzhou, China) for the mass spectrometry analysis.
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: firstname.lastname@example.org; email@example.com. These authors contributed equally to this work.
This study was supported by the National Natural Science Foundation of China (81670344, 81370421, 81370330) and the Natural Science Foundation of Heilongjiang (No. D201070).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 5.Fukushima A, Lopaschuk GD (2016) Acetylation control of cardiac fatty acid beta-oxidation and energy metabolism in obesity, diabetes, and heart failure. Biochim Biophys ActaGoogle Scholar
- 10.Lantier L, Williams AS, Williams IM, Yang KK, Bracy DP, Goelzer M, James FD, Gius D, Wasserman DH (2015) SIRT3 is crucial for maintaining skeletal muscle insulin action and protects against severe insulin resistance in high-fat-fed mice. Diabetes 64(9):3081–3092CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Vassilopoulos A, Pennington JD, Andresson T, Rees DM, Bosley AD, Fearnley IM, Ham A, Flynn CR, Hill S, Rose KL, Kim HS, Deng CX, Walker JE, Gius D (2014) SIRT3 deacetylates ATP synthase F1 complex proteins in response to nutrient- and exercise-induced stress. Antioxid Redox Signal 21(4):551–564CrossRefPubMedPubMedCentralGoogle Scholar
- 34.Revollo JR, Korner A, Mills KF, Satoh A, Wang T, Garten A, Dasgupta B, Sasaki Y, Wolberger C, Townsend RR, Milbrandt J, Kiess W, Imai S (2007) Nampt/PBEF/Visfatin regulates insulin secretion in beta cells as a systemic NAD biosynthetic enzyme. Cell Metab 6(5):363–375CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Alrob OA, Sankaralingam S, Ma C, Wagg CS, Fillmore N, Jaswal JS, Sack MN, Lehner R, Gupta MP, Michelakis ED, Padwal RS, Johnstone DE, Sharma AM, Lopaschuk GD (2014) Obesity-induced lysine acetylation increases cardiac fatty acid oxidation and impairs insulin signalling. Cardiovasc Res 103(4):485–497CrossRefPubMedPubMedCentralGoogle Scholar
- 38.Peterson LR, Saeed IM, McGill JB, Herrero P, Schechtman KB, Gunawardena R, Recklein CL, Coggan AR, DeMoss AJ, Dence CS, Gropler RJ (2012) Sex and type 2 diabetes: obesity-independent effects on left ventricular substrate metabolism and relaxation in humans. Obesity (Silver Spring) 20(4):802–810CrossRefGoogle Scholar
- 40.Hirschey MD, Shimazu T, Goetzman E, Jing E, Schwer B, Lombard DB, Grueter CA, Harris C, Biddinger S, Ilkayeva OR, Stevens RD, Li Y, Saha AK, Ruderman NB, Bain JR, Newgard CB, Farese RV Jr, Alt FW, Kahn CR, Verdin E (2010) SIRT3 regulates mitochondrial fatty-acid oxidation by reversible enzyme deacetylation. Nature 464(7285):121–125CrossRefPubMedPubMedCentralGoogle Scholar
- 41.Jing E, O'Neill BT, Rardin MJ, Kleinridders A, Ilkeyeva OR, Ussar S, Bain JR, Lee KY, Verdin EM, Newgard CB, Gibson BW, Kahn CR (2013) Sirt3 regulates metabolic flexibility of skeletal muscle through reversible enzymatic deacetylation. Diabetes 62(10):3404–3417CrossRefPubMedPubMedCentralGoogle Scholar
- 43.Hirschey MD, Shimazu T, Jing E, Grueter CA, Collins AM, Aouizerat B, Stancakova A, Goetzman E, Lam MM, Schwer B, Stevens RD, Muehlbauer MJ, Kakar S, Bass NM, Kuusisto J, Laakso M, Alt FW, Newgard CB, Farese RV Jr, Kahn CR, Verdin E (2011) SIRT3 deficiency and mitochondrial protein hyperacetylation accelerate the development of the metabolic syndrome. Mol Cell 44(2):177–190CrossRefPubMedPubMedCentralGoogle Scholar
- 46.Bharathi SS, Zhang Y, Mohsen AW, Uppala R, Balasubramani M, Schreiber E, Uechi G, Beck ME, Rardin MJ, Vockley J, Verdin E, Gibson BW, Hirschey MD, Goetzman ES (2013) Sirtuin 3 (SIRT3) protein regulates long-chain acyl-CoA dehydrogenase by deacetylating conserved lysines near the active site. J Biol Chem 288(47):33837–33847CrossRefPubMedPubMedCentralGoogle Scholar