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The Klotho protein supports redox balance and metabolic functions of cardiomyocytes during ischemia/reperfusion injury
, 1, 1, 1
Affiliations- Division of Clinical Chemistry and Laboratory Haematology, Department of Medical Laboratory Diagnostics, Faculty of Pharmacy, Wroclaw Medical University, Wroclaw, Poland
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Abstract
Background: Acute heart ischemia followed by reperfusion leads to overproduction of reactive oxygen/ /nitrogen species (ROS/RNS), disrupted expression of nitric oxide synthase (NOS) and unbalanced glucose metabolism. Klotho is a membrane-bound or soluble protein that exerts protective activity in many organs. While Klotho is produced mainly in the kidneys and brain, it has been recently proven that Klotho is expressed in the cardiomyocytes as well. This study aimed to show the influence of the Klotho protein on oxidative/nitrosative stress and metabolic function of the cardiomyocytes subjected to ischemia/reperfusion (I/R) injury.
Methods: Human cardiac myocytes underwent in vitro chemical I/R (with sodium cyanide and 2-deoxyglucose), in the presence or absence of the recombinant human Klotho protein. The present study included an investigation of cell injury markers, level of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX), level of oxidative/nitrosative stress and metabolic processes of the cardiomyocytes.
Results: Administration of Klotho protein resulted in mitigation of injury, decreased level of NOX2 and NOX4, reduced generation of ROS/RNS and hydrogen peroxide (H2O2), decreased expression of inducible NOS and limited production of nitrates/nitrites in cells under I/R. Glucose uptake and lactate production in the cardiomyocytes subjected to I/R were normalized after Klotho supplementation.
Conclusions: The Klotho protein participates in the regulation of redox balance and supports metabolic homeostasis of the cardiomyocytes and hence, contributes to protection against I/R injury.
Abstract
Background: Acute heart ischemia followed by reperfusion leads to overproduction of reactive oxygen/ /nitrogen species (ROS/RNS), disrupted expression of nitric oxide synthase (NOS) and unbalanced glucose metabolism. Klotho is a membrane-bound or soluble protein that exerts protective activity in many organs. While Klotho is produced mainly in the kidneys and brain, it has been recently proven that Klotho is expressed in the cardiomyocytes as well. This study aimed to show the influence of the Klotho protein on oxidative/nitrosative stress and metabolic function of the cardiomyocytes subjected to ischemia/reperfusion (I/R) injury.
Methods: Human cardiac myocytes underwent in vitro chemical I/R (with sodium cyanide and 2-deoxyglucose), in the presence or absence of the recombinant human Klotho protein. The present study included an investigation of cell injury markers, level of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX), level of oxidative/nitrosative stress and metabolic processes of the cardiomyocytes.
Results: Administration of Klotho protein resulted in mitigation of injury, decreased level of NOX2 and NOX4, reduced generation of ROS/RNS and hydrogen peroxide (H2O2), decreased expression of inducible NOS and limited production of nitrates/nitrites in cells under I/R. Glucose uptake and lactate production in the cardiomyocytes subjected to I/R were normalized after Klotho supplementation.
Conclusions: The Klotho protein participates in the regulation of redox balance and supports metabolic homeostasis of the cardiomyocytes and hence, contributes to protection against I/R injury.
Keywords
Klotho protein, cardioprotection, heart, ischemia/reperfusion injury, oxidative stress, nitrosative stress
About this articleTitle
The Klotho protein supports redox balance and metabolic functions of cardiomyocytes during ischemia/reperfusion injury
Journal
Issue
Article type
Original Article
Pages
836-849
Published online
2021-12-28
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4756
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936
DOI
Pubmed
Bibliographic record
Cardiol J 2022;29(5):836-849.
Keywords
Klotho protein
cardioprotection
heart
ischemia/reperfusion injury
oxidative stress
nitrosative stress
Authors
Agnieszka Olejnik
Marta Banaszkiewicz
Anna Krzywonos-Zawadzka
Iwona Bil-Lula
References (52)- Heusch G, Gersh BJ. The pathophysiology of acute myocardial infarction and strategies of protection beyond reperfusion: a continual challenge. Eur Heart J. 2017; 38(11): 774–784.
- Byrne JA, Grieve DJ, Cave AC, et al. Oxidative stress and heart failure. Arch Mal Coeur Vaiss. 2003; 96(3): 214–221.
- Krzywonos-Zawadzka A, Franczak A, Olejnik A, et al. Cardioprotective effect of MMP-2-inhibitor-NO-donor hybrid against ischaemia/reperfusion injury. J Cell Mol Med. 2019; 23(4): 2836–2848.
- Tsutsui H, Kinugawa S, Matsushima S. Oxidative stress and heart failure. Am J Physiol Heart Circ Physiol. 2011; 301(6): H2181–H2190.
- Kuro-o M, Matsumura Y, Aizawa H, et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature. 1997; 390(6655): 45–51.
- Kim JH, Hwang KH, Park KS, et al. Biological role of anti-aging protein Klotho. J Lifestyle Med. 2015; 5(1): 1–6.
- Olejnik A, Krzywonos-Zawadzka A, Banaszkiewicz M, et al. Klotho protein contributes to cardioprotection during ischaemia/reperfusion injury. J Cell Mol Med. 2020; 24(11): 6448–6458.
- Lopaschuk GD, Stanley WC. Glucose metabolism in the ischemic heart. Circulation. 1997; 95(2): 313–315.
- Stanley WC, Hall JL, Stone CK, et al. Acute myocardial ischemia causes a transmural gradient in glucose extraction but not glucose uptake. Am J Physiol. 1992; 262(1 Pt 2): H91–H96.
- Stasiak P, Sznitowska M. Zastosowanie hodowli komórkowych w badaniach biofarmaceutycznych. Farm Pol. 2010; 66(3): 228–234.
- Lindsey ML, Bolli R, Canty JM, et al. Guidelines for experimental models of myocardial ischemia and infarction. Am J Physiol Heart Circ Physiol. 2018; 314(4): H812–H838.
- Lim SW, Jin L, Luo K, et al. Klotho enhances FoxO3-mediated manganese superoxide dismutase expression by negatively regulating PI3K/AKT pathway during tacrolimus-induced oxidative stress. Cell Death Dis. 2017; 8(8): e2972.
- Kalyanaraman B, Darley-Usmar V, Davies KJA, et al. Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations. Free Radic Biol Med. 2012; 52(1): 1–6.
- Bil-Lula I, Lin HB, Biały D, et al. Subthreshold nitric oxide synthase inhibition improves synergistic effects of subthreshold MMP-2/MLCK-mediated cardiomyocyte protection from hypoxic injury. J Cell Mol Med. 2016; 20(6): 1086–1094.
- Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72(1-2): 248–254.
- Sugiura H, Yoshida T, Tsuchiya K, et al. Klotho reduces apoptosis in experimental ischaemic acute renal failure. Nephrol Dial Transplant. 2005; 20(12): 2636–2645.
- Hu MC, Shi M, Zhang J, et al. Klotho deficiency is an early biomarker of renal ischemia-reperfusion injury and its replacement is protective. Kidney Int. 2010; 78(12): 1240–1251.
- Hu MC, Shi M, Gillings N, et al. Recombinant alpha-Klotho may be prophylactic and therapeutic for acute to chronic kidney disease progression and uremic cardiomyopathy. Kidney Int. 2017; 91(5): 1104–1114.
- Qian Y, Guo X, Che L, et al. Klotho Reduces Necroptosis by Targeting Oxidative Stress Involved in Renal Ischemic-Reperfusion Injury. Cell Physiol Biochem. 2018; 45(6): 2268–2282.
- Zhou HJ, Li H, Shi MQ, et al. Protective effect of Klotho against ischemic brain injury is associated with inhibition of rig-i/nf-kappaB signaling. Front Pharmacol. 2017; 8: 950.
- Marçais C, Maucort-Boulch D, Drai J, et al. Circulating klotho associates with cardiovascular morbidity and mortality during hemodialysis. J Clin Endocrinol Metab. 2017; 102(9): 3154–3161.
- Paula RS, Souza VC, Machado-Silva W, et al. Serum Klotho (but not haplotypes) associate with the post-myocardial infarction status of older adults. Clinics (Sao Paulo). 2016; 71(12): 725–732.
- Sahu A, Mamiya H, Shinde SN, et al. Age-related declines in α-Klotho drive progenitor cell mitochondrial dysfunction and impaired muscle regeneration. Nat Commun. 2018; 9(1): 4859.
- Kuroda J, Ago T, Matsushima S, et al. NADPH oxidase 4 (Nox4) is a major source of oxidative stress in the failing heart. Proc Natl Acad Sci U S A. 2010; 107(35): 15565–15570.
- Bedard K, Krause KH. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 2007; 87(1): 245–313.
- Dikalov SI, Dikalova AE, Bikineyeva AT, et al. Distinct roles of Nox1 and Nox4 in basal and angiotensin ii-stimulated superoxide and hydrogen peroxide production. Free Radic Biol Med. 2008; 45(9): 1340–1351.
- Simone S, Rascio F, Castellano G, et al. Complement-dependent NADPH oxidase enzyme activation in renal ischemia/reperfusion injury. Free Radic Biol Med. 2014; 74: 263–273.
- Yang Ke, Wang C, Nie L, et al. Klotho protects against indoxyl sulphate-induced myocardial hypertrophy. J Am Soc Nephrol. 2015; 26(10): 2434–2446.
- Krijnen PAJ, Meischl C, Hack CE, et al. Increased Nox2 expression in human cardiomyocytes after acute myocardial infarction. J Clin Pathol. 2003; 56(3): 194–199.
- Li JM, Gall NP, Grieve DJ, et al. Activation of NADPH oxidase during progression of cardiac hypertrophy to failure. Hypertension. 2002; 40(4): 477–484.
- Wang Y, Kuro-o M, Sun Z. Klotho gene delivery suppresses Nox2 expression and attenuates oxidative stress in rat aortic smooth muscle cells via the cAMP-PKA pathway. Aging Cell. 2012; 11(3): 410–417.
- Takenaka T, Kobori H, Inoue T, et al. [op.4b.02] Klotho supplementation attenuates blood pressure and oxidative stress in diabetes. J Hypertens. 2017; 35(Suppl 2): e38.
- Yamamoto M, Clark JD, Pastor JV, et al. Regulation of oxidative stress by the anti-aging hormone klotho. J Biol Chem. 2005; 280(45): 38029–38034.
- Mitobe M, Yoshida T, Sugiura H, et al. Oxidative stress decreases klotho expression in a mouse kidney cell line. Nephron Exp Nephrol. 2005; 101(2): e67–e74.
- Heusch P, Aker S, Boengler K, et al. Increased inducible nitric oxide synthase and arginase II expression in heart failure: no net nitrite/nitrate production and protein S-nitrosylation. Am J Physiol Heart Circ Physiol. 2010; 299(2): H446–H453.
- Yasmin W, Strynadka KD, Schulz R. Generation of peroxynitrite contributes to ischemia-reperfusion injury in isolated rat hearts. Cardiovasc Res. 1997; 33(2): 422–432.
- Krzywonos-Zawadzka A, Wozniak M, Sawicki G, et al. A drug cocktail for protecting against ischemia-reperfusion injury. Front Biosci (Landmark Ed). 2020; 25: 722–735.
- Krzywonos-Zawadzka A, Franczak A, Sawicki G, et al. Mixture of MMP-2, MLC, and NOS Inhibitors Affects NO Metabolism and Protects Heart from Cardiac I/R Injury. Cardiol Res Pract. 2020; 2020: 1561478.
- Bil-Lula I, Krzywonos-Zawadzka A, Sawicka J, et al. L-NAME improves doxycycline and ML-7 cardioprotection from oxidative stress. Front Biosci (Landmark Ed). 2018; 23: 298–309.
- Corsetti G, Pasini E, Scarabelli TM, et al. Decreased expression of Klotho in cardiac atria biopsy samples from patients at higher risk of atherosclerotic cardiovascular disease. J Geriatr Cardiol. 2016; 13(8): 701–711.
- Saito Y, Nakamura T, Ohyama Y, et al. In vivo klotho gene delivery protects against endothelial dysfunction in multiple risk factor syndrome. Biochem Biophys Res Commun. 2000; 276(2): 767–772.
- Saito Y, Yamagishi T, Nakamura T, et al. Klotho protein protects against endothelial dysfunction. Biochem Biophys Res Commun. 1998; 248(2): 324–329.
- Liu Y, Zhang Q. Periodontitis aggravated pancreatic beta-cell dysfunction in diabetic mice through interleukin-12 regulation on Klotho. J Diabetes Investig. 2016; 7(3): 303–311.
- Yang Ke, Nie L, Huang Y, et al. Amelioration of uremic toxin indoxyl sulfate-induced endothelial cell dysfunction by Klotho protein. Toxicol Lett. 2012; 215(2): 77–83.
- Neely JR, Grotyohann LW. Role of glycolytic products in damage to ischemic myocardium. Dissociation of adenosine triphosphate levels and recovery of function of reperfused ischemic hearts. Circ Res. 1984; 55(6): 816–824.
- Zhang Y, Liu G, Gao X. Attenuation of protects cardiomyocytes against hypoxic stress through maintenance of glycolysis. Biosci Rep. 2017; 37(6).
- Liao R, Jain M, Cui L, et al. Cardiac-specific overexpression of GLUT1 prevents the development of heart failure attributable to pressure overload in mice. Circulation. 2002; 106(16): 2125–2131.
- Malhotra R, Tyson DGW, Sone H, et al. Glucose uptake and adenoviral mediated GLUT1 infection decrease hypoxia-induced HIF-1alpha levels in cardiac myocytes. J Mol Cell Cardiol. 2002; 34(8): 1063–1073.
- Malhotra R, Brosius FC. Glucose uptake and glycolysis reduce hypoxia-induced apoptosis in cultured neonatal rat cardiac myocytes. J Biol Chem. 1999; 274(18): 12567–12575.
- Utsugi T, Ohno T, Ohyama Y, et al. Decreased insulin production and increased insulin sensitivity in the klotho mutant mouse, a novel animal model for human aging. Metabolism. 2000; 49(9): 1118–1123.
- Lin Yi, Sun Z. Antiaging gene Klotho enhances glucose-induced insulin secretion by up-regulating plasma membrane levels of TRPV2 in MIN6 beta-cells. Endocrinology. 2012; 153(7): 3029–3039.
- Donate-Correa J, Martín-Núñez E, Delgado NP, et al. Implications of fibroblast growth factor/Klotho system in glucose metabolism and diabetes. Cytokine Growth Factor Rev. 2016; 28: 71–77.
