Dexmedetomidine improves myocardial ischemia-reperfusion injury by increasing autophagy via PINK1/PRKN pathway

Ischemic heart disease poses a great threat to human life with its high morbidity and mortality. Timely reperfusion is considered as the most effective intervention for clinical treatment of myocardial ischemia. Reperfusion can commonly lead to cell death and myocardial injury. There has already been evidence that dexmedetomidine (DEX) can protect the injury of myocardial ischemia reperfusion (MI/R). The aims of this research are to explain DEX’s functions in protecting MI/R injury as well as probing into its inner mechanisms. H9c2 cells was adopted to generate the injury of MI/R cell as a result of hypoxia/reoxygenation (H/R). MTT test was used to examine cell activity and flow cytometry was utilized to check cell apoptosis rate. Western blot test was adopted to appraise the protein expressions of Parkin RBR E3 ubiquitin protein ligase (PRKN or Parkin), PTEN-induced kinase-1 (PINK1) and markers associate with autophagy (Light Chain 3-II/I (LC3-II/I)). Immunofluorescence was applied to estimate the LC3 level. Next, the function of DEX in protecting the injury of H/R induced cell by PINK1/PRKN pathway was confirmed. DEX treatment significantly promoted viability, improved apoptotic rate and increased autophagy in H9c2 cells processed by H/R. Moreover, the protein expressions of PINK1 and PRKN were boosted by DEX, suggesting that DEX induced autophagy through the activation of PINK1/PRKN signaling pathway. In later tests, PINK1 teardown weakened DEX’s effect the injury of myocardial cell induced by H/R. DEX improves MI/R injury by inducing autophagy through activating PINK1/PRKN pathway.

Myocardial ischemia-reperfusion; Dexmedetomidine; Autophagy; Apoptosis; PINK1

[1] Gewirtz H, Dilsizian V. Myocardial viability: survival mechanisms and molecular imaging targets in acute and chronic ischemia. Circulation Research. 2017; 120: 1197–1212.

[2] Heusch G. Myocardial ischaemia–reperfusion injury and cardioprotection in perspective. Nature Reviews Cardiology. 2020; 17: 773–789.

[3] Gazmuri RJ, Ayoub IM, Radhakrishnan J. Myocardial effects of cardiac arrest and resuscitation with especial reference to mitochondrial injury. Signa Vitae. 2008; 3: 7–12.

[4] Powers SK, Smuder AJ, Kavazis AN, Quindry JC. Mechanisms of exercise-induced cardioprotection. Physiology. 2014; 29: 27–38.

[5] Du J, Li YL, Zhao W. Autophagy and myocardial ischemia. Advances in Experimental Medicine and Biology. 2020; 32: 217–222.

[6] Doherty J, Baehrecke EH. Life, death and autophagy. Nature Cell Biology. 2018; 20: 1110–1117.

[7] Wu SY, Chang GL, Gao L, Jiang D, Wang LY, Li GX, et al. Trimetazidine protects against myocardial ischemia/reperfusion injury by inhibiting excessive autophagy. Journal of Molecular Medicine. 2018; 96: 791–806.

[8] Mo YL, Tang L, Ma Y, Wu SZ. Pramipexole pretreatment attenuates myocardial ischemia/reperfusion injury through upregulation of autophagy. Biochemical and Biophysical Research Communications. 2016; 473: 1119–1124.

[9] Kawazoe Y, Miyamoto K, Morimoto T, Yamamoto T, Fuke A, Hashimoto A, et al. Effect of dexmedetomidine on mortality and ventilator-free days in patients requiring mechanical ventilation with sepsis: a randomized clinical trial. Journal of the American Medical Association. 2017; 317: 1321–1328.

[10] Sun ZX, Zhao TY, Lv SJ, Gao Y, Masters J, Weng H. Dexmedetomidine attenuates spinal cord ischemia–reperfusion injury through both anti-inflammation and anti-apoptosis mechanisms in rabbits. Journal of Translational Medicine. 2018; 16: 209.

[11] Zhang X, Li YX, Wang Y, Zhuang YR, Ren XJ, Yang K, et al. Dexmedetomidine postconditioning suppresses myocardial ischemia/reperfusion injury by activating the SIRT1/mTOR axis. Bioscience Reports. 2020; 40: BSR20194030.

[12] Yu TY, Liu D, Gao M, Yang PL, Zhang M, Song F, et al. Dexmedetomidine prevents septic myocardial dysfunction in rats via activation of α7nAChR and PI3K/Akt- mediated autophagy. Biomedicine & Pharmacotherapy. 2019; 120: 109231.

[13] Pickrell AM, Youle RJ. The roles of PINK1, parkin, and mitochondrial fidelity in parkinson’s disease. Neuron. 2015; 85: 257–273.

[14] Lazarou M, Jin SM, Kane LA, Youle RJ. Role of PINK1 Binding to the TOM complex and alternate intracellular membranes in recruitment and activation of the E3 ligase parkin. Developmental Cell. 2012; 22: 320–333.

[15] Eiyama A, Okamoto K. PINK1/Parkin-mediated mitophagy in mammalian cell. Current Opinion in Cell Biology. 2015; 33: 95–101.

[16] Wang YY, Mao X, Chen HG, Feng JC, Yan MY, Wang YQ, et al. Dexmedetomidine alleviates LPS-induced apoptosis and inflammation in macrophages by eliminating damaged mitochondria via PINK1 mediated mitophagy. International Immunopharmacology. 2019; 73: 471–481.

[17] Liu WH, Chen CG, Gu XJ, Zhang L, Mao X, Chen ZL, et al. AM1241 alleviates myocardial ischemia-reperfusion injury in rats by enhancing Pink1/Parkin-mediated autophagy. Life Sciences. 2021; 272: 119228.

[18] Qiu Z, He YH, Ming H, Lei SQ, Leng Y, Xia ZY. Lipopolysaccharide (LPS) aggravates high glucose- and hypoxia/reoxygenation-induced injury through activating ROS-Dependent NLRP3 inflammasome-mediated pyroptosis in H9C2 cardiomyocytes. Journal of Diabetes Research. 2019; 2019: 8151836.

[19] Tang L, Mo YL, Li YP, Zhong YK, He SF, Zhang Y, et al. Urolithin a alleviates myocardial ischemia/reperfusion injury via PI3K/Akt pathway. Biochemical and Biophysical Research Communications. 2017; 486: 774–780.

[20] Shang ZF, Wei Q, Yu L, Huang F, Xiao BB, Wang H, et al. Suppression of PC-1/PrLZ sensitizes prostate cancer cells to ionizing radiation by attenuating DNA damage repair and inducing autophagic cell death. Oncotarget. 2016; 7: 62340-62351.

[21] Zheng X, Zhang J, Li S, Gao X, Zhang Y, Wang M, et al. Low doses of niclosamide and quinacrine combination yields synergistic effect in melanoma via activating autophagy-mediated p53-dependent apoptosis. Translational Oncology. 2022; 21:101425.

[22] Korshunova AY, Blagonravov ML, Neborak EV, Syatkin SP, Sklifasovskaya AP, Semyatov SM, et al. BCL2-regulated apoptotic process in myocardial ischemia-reperfusion injury. International Journal of Molecular Medicine. 2021; 47: 23–26.

[23] Frank A, Bonney M, Bonney S, Weitzel L, Koeppen M, Eckle T. Myocardial ischemia reperfusion injury: from basic science to clinical bedside. Seminars in Cardiothoracic and Vascular Anesthesia. 2012; 16: 123–132.

[24] Afonso J, Reis F. Dexmedetomidine: current role in anesthesia and intensive care. Brazilian Journal of Anesthesiology. 2012; 62: 118–133.

[25] Wang HK, Xia Y, Sun ZQ. Dexmedetomidine: a potential therapy for myocardial ischemia/reperfusion injury. International Journal of Cardiology. 2021; 335: 104.

[26] Huang Y, Sun XT, Juan ZD, Zhang R, Wang RG, Meng SQ, et al. Dexmedetomidine attenuates myocardial ischemia-reperfusion injury in vitro by inhibiting NLRP3 Inflammasome activation. BioMed Central Anesthesiology. 2021; 21: 104.

[27] Yang YF WH, Song N, Jiang YH, Zhang J, Meng XW, et al. Dexmedetomidine attenuates ischemia/reperfusion-induced myocardial inflammation and apoptosis through inhibiting endoplasmic reticulum stress signaling. Journal of Inflammation Research. 2021; 14: 1217–1233.

[28] Meyer G, Czompa A, Reboul C, Csepanyi E, Czegledi A, Bak I, et al. The cellular autophagy markers Beclin-1 and LC3B-II are increased during reperfusion in fibrillated mouse hearts. Current Pharmaceutical Design. 2013; 19: 6912–6918.

[29] Gustafsson AB, Gottlieb RA. Autophagy in ischemic heart disease. Circulation Research. 2009; 104: 150–158.

[30] Matsui Y, Takagi H, Qu X, Abdellatif M, Sakoda H, Asano T, et al. Distinct roles of autophagy in the heart during ischemia and reperfusion : roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy. Circulation Research. 2007; 100: 914–922.

[31] Zhao R, Xie E, Yang XB, Gong B. Alliin alleviates myocardial ischemia-reperfusion injury by promoting autophagy. Biochemical and Biophysical Research Communications. 2019; 512: 236–243.