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Remote Ischemic Conditioning Mediates Cardio-protection After Myocardial Ischemia/Reperfusion Injury by Reducing 4-HNE Levels and Regulating Autophagy via the ALDH2/SIRT3/HIF1α Signaling Pathway

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Abstract

Remote ischemic conditioning (RIC) can be effectively applied for cardio-protection. Here, to clarify whether RIC exerts myocardial protection via aldehyde dehydrogenase 2 (ALDH2), we established a myocardial ischemia/reperfusion (I/R) model in C57BL/6 and ALDH2 knockout (ALDH2-KO) mice and treated them with RIC. Echocardiography and single-cell contraction experiments showed that RIC significantly improved myocardial function and alleviated I/R injury in C57BL/6 mice but did not exhibit its cardioprotective effects in ALDH2-KO mice. TUNEL, Evan’s blue/triphenyl tetrazolium chloride, and reactive oxygen species (ROS) assays showed that RIC’s effect on reducing myocardial cell apoptosis, myocardial infarction area, and ROS levels was insignificant in ALDH2-KO mice. Our results showed that RIC could increase ALDH2 protein levels, activate sirtuin 3 (SIRT3)/hypoxia-inducible factor 1-alpha (HIF1α), inhibit autophagy, and exert myocardial protection. This study revealed that RIC could exert myocardial protection via the ALDH2/SIRT3/HIF1α signaling pathway by reducing 4-HNE secretion.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Code Availability

Not applicable.

Abbreviations

RIC:

Remote ischemic conditioning

ALDH2:

Aldehyde dehydrogenase 2

I/R:

Ischemia/reperfusion

ROS:

Reactive oxygen species

SIRT3:

Sirtuin 3

HIF1α:

Hypoxia-inducible factor 1-alpha

4-HNE:

4-hydroxynonenal

ALDH2-KO:

ALDH2 knockout

TTC:

1% 2,3,5-triphenyl tetrazolium chloride

PS:

Peak shortening

-dL/dt:

Maximal velocity of shortening

DHE:

Dihydroethidium

LVFS:

Left ventricular fractional shortening

LVEF:

Left ventricular ejection fraction

AAR:

Area at risk

WT:

Wild-type

References

  1. Heusch G. Myocardial ischaemia-reperfusion injury and cardioprotection in perspective. Nat Rev Cardiol. 2020;17:773–89.

    Article  PubMed  Google Scholar 

  2. Davidson SM, Ferdinandy P, Andreadou I, et al. Multitarget strategies to reduce myocardial ischemia/reperfusion injury: JACC review topic of the week. J Am Coll Cardiol. 2019;73:89–99.

    Article  PubMed  Google Scholar 

  3. Hirschhäuser C, Lissoni A, Görge PM, et al. Connexin 43 phosphorylation by casein kinase 1 is essential for the cardioprotection by ischemic preconditioning. Basic Res Cardiol. 2021;116:21.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Hong DM, Lee EH, Kim HJ, et al. Does remote ischaemic preconditioning with postconditioning improve clinical outcomes of patients undergoing cardiac surgery? Remote Ischaemic Preconditioning with Postconditioning Outcome Trial. Eur Heart J. 2014;35:176–83.

    Article  PubMed  Google Scholar 

  5. Wei X, Wu B, Zhao J, et al. Myocardial hypertrophic preconditioning attenuates cardiomyocyte hypertrophy and slows progression to heart failure through upregulation of S100A8/A9. Circulation. 2015;131:1506–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Skyschally A, Gent S, Amanakis G, et al. Across-species transfer of protection by remote ischemic preconditioning with species-specific myocardial signal transduction by reperfusion injury salvage kinase and survival activating factor enhancement pathways. Circ Res. 2015;117:279–88.

    Article  CAS  PubMed  Google Scholar 

  7. Amanakis G, Kleinbongard P, Heusch G, et al. Attenuation of ST-segment elevation after ischemic conditioning maneuvers reflects cardioprotection online. Basic Res Cardiol. 2019;114:22.

    Article  PubMed  Google Scholar 

  8. Zarbock A, Schmidt C, Van Aken H, et al. Effect of remote ischemic preconditioning on kidney injury among high-risk patients undergoing cardiac surgery: a randomized clinical trial. JAMA. 2015;313:2133–41.

    Article  CAS  PubMed  Google Scholar 

  9. Chen CH, Ferreira JC, Gross ER, et al. Targeting aldehyde dehydrogenase 2: new therapeutic opportunities. Physiol Rev. 2014;94:1–34.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Gross ER, Zambelli VO, Small BA, et al. A personalized medicine approach for Asian Americans with the aldehyde dehydrogenase 2*2 variant. Annu Rev Pharmacol Toxicol. 2015;55:107–27.

    Article  CAS  PubMed  Google Scholar 

  11. Ma LL, Ding ZW, Yin PP, et al. Hypertrophic preconditioning cardioprotection after myocardial ischaemia/reperfusion injury involves ALDH2-dependent metabolism modulation. Redox Biol. 2021;43:101960.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Chen CH, Budas GR, Churchill EN, et al. Activation of aldehyde dehydrogenase-2 reduces ischemic damage to the heart. Science. 2008;321:1493–5.

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ueta CB, Campos JC, Albuquerque RPE, et al. Cardioprotection induced by a brief exposure to acetaldehyde: role of aldehyde dehydrogenase 2. Cardiovasc Res. 2018;114:1006–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Zhang ZX, Li H, He JS, et al. Remote ischemic postconditioning alleviates myocardial ischemia/reperfusion injury by up-regulating ALDH2. Eur Rev Med Pharmacol Sci. 2018;22:6475–84.

    PubMed  Google Scholar 

  15. Yu Y, Jia XJ, Zong QF, et al. Remote ischemic postconditioning protects the heart by upregulating ALDH2 expression levels through the PI3K/Akt signaling pathway. Mol Med Rep. 2014;10:536–42.

    Article  CAS  PubMed  Google Scholar 

  16. Li CY, Ma W, Liu KP, et al. Advances in intervention methods and brain protection mechanisms of in situ and remote ischemic postconditioning. Metab Brain Dis. 2021;36:53–65.

    Article  CAS  PubMed  Google Scholar 

  17. Joseph B, Pandit V, Zangbar B, et al. Secondary brain injury in trauma patients: the effects of remote ischemic conditioning. J Trauma Acute Care Surg. 2015;78:698–703.

    Article  PubMed  Google Scholar 

  18. Kitagawa K, Kawamoto T, Kunugita N, et al. Aldehyde dehydrogenase (ALDH) 2 associates with oxidation of methoxyacetaldehyde; in vitro analysis with liver subcellular fraction derived from human and Aldh2 gene targeting mouse. FEBS Lett. 2000;476:306–11.

    Article  CAS  PubMed  Google Scholar 

  19. Sun X, Gao R, Li W, et al. Alda-1 treatment promotes the therapeutic effect of mitochondrial transplantation for myocardial ischemia-reperfusion injury. Bioact Mater. 2021;6:2058–69.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Gao RF, Li X, Xiang HY, et al. The covalent NLRP3-inflammasome inhibitor Oridonin relieves myocardial infarction induced myocardial fibrosis and cardiac remodeling in mice. Int Immunopharmacol. 2021;90:107133.

    Article  CAS  PubMed  Google Scholar 

  21. Ackers-Johnson M, Li PY, Holmes AP, et al. A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart. Circ Res. 2016;119:909–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Fan Q, Tao R, Zhang H, et al. Dectin-1 contributes to myocardial ischemia/reperfusion injury by regulating macrophage polarization and neutrophil infiltration. Circulation. 2019;139:663–78.

    Article  CAS  PubMed  Google Scholar 

  23. Si Y, Bao H, Han L, et al. Dexmedetomidine attenuation of renal ischaemia-reperfusion injury requires sirtuin 3 activation. Br J Anaesth. 2018;121:1260–71.

    Article  CAS  PubMed  Google Scholar 

  24. Wang Z, Sun R, Wang G, et al. SIRT3-mediated deacetylation of PRDX3 alleviates mitochondrial oxidative damage and apoptosis induced by intestinal ischemia/reperfusion injury. Redox Biol. 2020;28:101343.

    Article  CAS  PubMed  Google Scholar 

  25. Lu Z, Bourdi M, Li JH, et al. SIRT3-dependent deacetylation exacerbates acetaminophen hepatotoxicity. EMBO Rep. 2011;12:840–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Stiermaier T, Jensen JO, Rommel KP, et al. Combined intrahospital remote ischemic perconditioning and postconditioning improves clinical outcome in ST-elevation myocardial infarction. Circ Res. 2019;124:1482–91.

    Article  CAS  PubMed  Google Scholar 

  27. Vaibhav K, Braun M, Khan MB, et al. Remote ischemic post-conditioning promotes hematoma resolution via AMPK-dependent immune regulation. J Exp Med. 2018;215:2636–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Heusch G. Molecular basis of cardioprotection: signal transduction in ischemic pre-, post-, and remote conditioning. Circ Res. 2015;116:674–99.

    Article  ADS  CAS  PubMed  Google Scholar 

  29. Tsibulnikov SY, Maslov LN, Gorbunov AS, et al. A review of humoral factors in remote preconditioning of the heart. J Cardiovasc Pharmacol Ther. 2019;24:403–21.

    Article  CAS  PubMed  Google Scholar 

  30. Hummitzsch L, Zitta K, Fritze L, et al. Effects of remote ischemic preconditioning (RIPC) and chronic remote ischemic preconditioning (cRIPC) on levels of plasma cytokines, cell surface characteristics of monocytes and in-vitro angiogenesis: a pilot study. Basic Res Cardiol. 2021;116:60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Gao J, Min F, Wang S, et al. Effect of Rho-kinase and autophagy on remote ischemic conditioning-induced cardioprotection in rat myocardial ischemia/reperfusion injury model. Cardiovasc Ther. 2022;2022:6806427.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Yan Z, Du L, Liu Q, et al. Remote limb ischaemic conditioning produces cardioprotection in rats with testicular ischaemia-reperfusion injury. Exp Physiol. 2021;106:2223–34.

    Article  CAS  PubMed  Google Scholar 

  33. Zhao Y, Ding M, Yan F, et al. Inhibition of the JAK2/STAT3 pathway and cell cycle re-entry contribute to the protective effect of remote ischemic pre-conditioning of rat hindlimbs on cerebral ischemia/reperfusion injury. CNS Neurosci Ther. 2022. https://doi.org/10.1111/cns.14023.

  34. Mi L, Zhang N, Wan J, et al. Remote ischemic post-conditioning alleviates ischemia/reperfusion-induced intestinal injury via the ERK signaling pathway-mediated RAGE/HMGB axis. Mol Med Rep. 2021:24.

  35. Ajoolabady A, Aslkhodapasandhokmabad H, Aghanejad A, et al. Mitophagy receptors and mediators: therapeutic targets in the management of cardiovascular ageing. Ageing Res Rev. 2020;62:101129.

    Article  CAS  PubMed  Google Scholar 

  36. Santin Y, Fazal L, Sainte-Marie Y, et al. Mitochondrial 4-HNE derived from MAO-A promotes mitoCa(2+) overload in chronic postischemic cardiac remodeling. Cell Death Differ. 2020;27:1907–23.

    Article  CAS  PubMed  Google Scholar 

  37. Beretta M, Wölkart G, Schernthaner M, et al. Vascular bioactivation of nitroglycerin is catalyzed by cytosolic aldehyde dehydrogenase-2. Circ Res. 2012;110:385–93.

    Article  CAS  PubMed  Google Scholar 

  38. Ma H, Guo R, Yu L, et al. Aldehyde dehydrogenase 2 (ALDH2) rescues myocardial ischaemia/reperfusion injury: role of autophagy paradox and toxic aldehyde. Eur Heart J. 2011;32:1025–38.

    Article  CAS  PubMed  Google Scholar 

  39. Zhai M, Li B, Duan W, et al. Melatonin ameliorates myocardial ischemia reperfusion injury through SIRT3-dependent regulation of oxidative stress and apoptosis. J Pineal Res. 2017;63.

  40. Harris PS, Gomez JD, Backos DS, et al. Characterizing sirtuin 3 deacetylase affinity for aldehyde dehydrogenase 2. Chem Res Toxicol. 2017;30:785–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Koeppen M, Lee JW, Seo SW, et al. Hypoxia-inducible factor 2-alpha-dependent induction of amphiregulin dampens myocardial ischemia-reperfusion injury. Nat Commun. 2018;9:816.

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  42. Cai Z, Luo W, Zhan H, et al. Hypoxia-inducible factor 1 is required for remote ischemic preconditioning of the heart. Proc Natl Acad Sci U S A. 2013;110:17462–7.

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  43. Kalakech H, Tamareille S, Pons S, et al. Role of hypoxia inducible factor-1α in remote limb ischemic preconditioning. J Mol Cell Cardiol. 2013;65:98–104.

    Article  CAS  PubMed  Google Scholar 

  44. Zhao Y, Wang B, Zhang J, et al. ALDH2 (aldehyde dehydrogenase 2) protects against hypoxia-induced pulmonary hypertension. Arterioscler Thromb Vasc Biol. 2019;39:2303–19.

    Article  CAS  PubMed  Google Scholar 

  45. Laitakari A, Ollonen T, Kietzmann T, et al. Systemic inactivation of hypoxia-inducible factor prolyl 4-hydroxylase 2 in mice protects from alcohol-induced fatty liver disease. Redox Biol. 2019;22:101145.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Ji W, Wei S, Hao P, et al. Aldehyde dehydrogenase 2 has cardioprotective effects on myocardial ischaemia/reperfusion injury via suppressing mitophagy. Front Pharmacol. 2016;7:101.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Kornfeld OS, Hwang S, Disatnik MH, et al. Mitochondrial reactive oxygen species at the heart of the matter: new therapeutic approaches for cardiovascular diseases. Circ Res. 2015;116:1783–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Guéraud F. 4-Hydroxynonenal metabolites and adducts in pre-carcinogenic conditions and cancer. Free Radic Biol Med. 2017;111:196–208.

    Article  PubMed  Google Scholar 

  49. Mak S, Lehotay DC, Yazdanpanah M, et al. Unsaturated aldehydes including 4-OH-nonenal are elevated in patients with congestive heart failure. J Card Fail. 2000;6:108–14.

    Article  CAS  PubMed  Google Scholar 

  50. Eng MY, Luczak SE, Wall TL. ALDH2, ADH1B, and ADH1C genotypes in Asians: a literature review. Alcohol Res Health. 2007;30:22–7.

    PubMed  PubMed Central  Google Scholar 

  51. Yukawa Y, Muto M, Hori K, et al. Combination of ADH1B*2/ALDH2*2 polymorphisms alters acetaldehyde-derived DNA damage in the blood of Japanese alcoholics. Cancer Sci. 2012;103:1651–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Li H, Borinskaya S, Yoshimura K, et al. Refined geographic distribution of the oriental ALDH2*504Lys (nee 487Lys) variant. Ann Hum Genet. 2009;73:335–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This research was funded by the Jiangxi Provincial Natural Science Foundation Project (20071BBG70067, 20181074), and the National Natural Science Foundation of China (81160019 and 81360031).

Author information

Author notes

  1. Rifeng Gao, Chunyu Lv, and Yanan Qu contributed equally.

Authors and Affiliations

  1. Department of Cardiovascular Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang University, Nanchang, China

    Rifeng Gao, Hen Yang, Chuangze Hao & Yanhua Tang

  2. Department of Cardiology, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, Shanghai, China

    Rifeng Gao & Yiqing Yang

  3. Shenzhen Key Laboratory for Neuronal Structural Biology, Biomedical Research Institute, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, China

    Chunyu Lv

  4. Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 20032, China

    Yanan Qu, Xiaolei Sun & Yanhua Tang

  5. First Affiliated Hospital of Zhejiang University, Hangzhou, China

    Xiaosheng Hu

Authors
  1. Rifeng Gao
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Contributions

Rifeng Gao, Chunyu Lv, and Yanan Qu designed the research and wrote the paper. Rifeng Gao, Heng Yang, Xiaolei Sun, and Chuangze Hao performed the experiments. Rifeng Gao, Xiaosheng Hu, Yiqing Yang, and Yanhua Tang contributed new reagents/analytic tools and provide critical suggestions. Rifeng Gao, Chunyu Lv, and Yanan Qu analyzed the data. Xiaosheng Hu, Yiqing Yang, and Yanhua Tang edited the paper.

Corresponding authors

Correspondence to Xiaosheng Hu, Yiqing Yang or Yanhua Tang.

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Animal Studies

All animal experiments were approved by the Animal Care Ethics Committee of Fudan University and performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. The mice were kept under a 12:12-h light/dark cycle at a consistent temperature and humidity and were also given ad libitum access to food and water. Additional dose of analgesics was given if the animals appeared to be experiencing pain (based on criteria such as immobility and failure to eat). At the indicated time points, mice were euthanized by CO2/cervical dislocation, and tissues were subsequently harvested for analyses.

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Associate Editor Yihua Bei oversaw the review of this article

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Supplementary Information

Supplementary Figure 1

RIC treatment preserves cardiac function after I/R. Representative images of echocardiography tracing in I/R mice treated with RIC in each group after 24 h reperfusion (a). Heart rate (BPM, n>6, b). Left ventricular ejection fraction (LVEF, n>6, c). Left ventricular fractional shortening (LVFS, n>6, D). Peak shortening (% cell lengthening, n>6, e). Maximal shortening velocity (-dl/dt, n>6, f). Data are depicted as the mean ± SEM. Statistical significance was determined by two-way ANOVA with a post-hoc Holm-Sidak test, ns, not significant; *P<0.05; ***P<0.001; compared with the control group. (PNG 1135 kb)

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Gao, R., Lv, C., Qu, Y. et al. Remote Ischemic Conditioning Mediates Cardio-protection After Myocardial Ischemia/Reperfusion Injury by Reducing 4-HNE Levels and Regulating Autophagy via the ALDH2/SIRT3/HIF1α Signaling Pathway. J. of Cardiovasc. Trans. Res. 17, 169–182 (2024). https://doi.org/10.1007/s12265-023-10355-z

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