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KCNJ11 Downregulation Stimulates Cardiac Cell Apoptosis in Myocarditis


Affiliations
1 Department of Life Sciences, Korea University, Seoul 02841, Korea, Republic of
2 Korea Basic Science Institute, Seoul Center, Seoul 02841, Korea, Republic of
3 Korea Basic Science Institute, Western Seoul Center, Seoul 03759, Korea, Republic of
 

One of the inflammatory heart diseases, viz. acute myocarditis, occurs due to cardiac cell death. However, the molecular mechanism underlying cell death remains largely unexplored. In this study, we report that the down-regulation of KCNJ11, a central subunit of the ATP-sensitive potassium (KATP) channel plays a key role in the reduction of blood glucose, and is involved in apoptosis of cardiac cells. Using proteomics analysis of experimental autoimmune myocarditis (EAM), we show that the KCNJ11 level marked by decreased, whereas Camk2a expression increased significantly in EAM tissues by 16 and 20 days, compared to control tissues. Using 1H-MAS NMR we also show that glucose levels were slightly elevated in EAM tissues. In vitro assays using H9c2 cardiac cells revealed that both lipopolysaccharide (LPS) and high glucose treatment decreased cell viability, in which toxicity was attenuated by treatment with KATP pharmacological openers, but not by the KATP blockers (Gli and 5-HD). Apoptosis induced by LPS or high glucose treatment was suppressed by Ca2+ chelator (BAPTA-AM) treatment. We found that KCNJ11 levels had decreased in cardiac cells by LPS or high glucose treatment, and siRNA-mediated knockdown of KCNJ11 expression further stimulated the LPS- or high glucose-induced apoptosis. Together, our results demonstrate first that KCNJ11 is down-regulated under inflammation and high glucose conditions and its inactivation facilitates cardiac cell apoptosis. We assume that down-regulation of KCNJ11 has an effect on the development of myocarditis.

Keywords

Apoptosis, Blood Glucose Lipopolysaccharide, Cardiac Cells, Myocarditis.
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  • Andreoletti, L., Leveque, N., Boulagnon, C., Brasselet, C. and Fornes, P., Viral causes of human myocarditis. Arch. Cardiovasc. Dis., 2009, 102, 559–568.

  • Pollack, A., Kontorovich, A. R., Fuster, V. and Dec, G. W., Viral myocarditis – diagnosis, treatment options, and current controversies. Nature Rev. Cardiol., 2015, 12, 670–680.

  • Smith, S. C., Autoimmune myocarditis. Current Protocols in Immunology, 2001, Chapter 15, Unit 15 14, 15.14.1–15.14.19.

  • Kodama, M. et al., Characteristics of giant cells and factors related to the formation of giant cells in myocarditis. Circ. Res., 1991, 69, 1042–1050.

  • Rose, N. R. and Hill, S. L., Autoimmune myocarditis. Int. J. Cardiol., 1996, 54, 171–175.

  • Hu, F. et al., Effects of 1, 25-dihydroxyvitamin D3 on experimental autoimmune myocarditis in mice. Cell Physiol. Biochem., 2016, 38, 2219–2229.

  • Iurlaro, R. and Munoz-Pinedo, C., Cell death induced by endoplasmic reticulum stress. FEBS J., 2016, 283, 2640–2652.

  • Gogvadze, V., Orrenius, S. and Zhivotovsky, B., Multiple pathways of cytochrome c release from mitochondria in apoptosis. Biochim. Biophys. Acta, 2006, 1757, 639–647.

  • Chen, Q., Gong, B. and Almasan, A., Distinct stages of cytochrome c release from mitochondria: evidence for a feedback amplification loop linking caspase activation to mitochondrial dysfunction in genotoxic stress induced apoptosis. Cell Death Differ., 2000, 7, 227–233.

  • Rizzuto, R. et al., Calcium and apoptosis: facts and hypotheses. Oncogene, 2003, 22, 8619–8627.

  • Pinton, P., Giorgi, C., Siviero, R., Zecchini, E. and Rizzuto, R., Calcium and apoptosis: ER–mitochondria Ca2+ transfer in the control of apoptosis. Oncogene, 2008, 27, 6407–6418.

  • Yu, T., Sheu, S. S., Robotham, J. L. and Yoon, Y., Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc. Res., 2008, 79, 341–351.

  • Chen, M., Wang, W., Ma, J., Ye, P. and Wang, K., High glucose induces mitochondrial dysfunction and apoptosis in human retinal pigment epithelium cells via promoting SOCS1 and Fas/FasL signaling. Cytokine, 2016, 78, 94–102.

  • La-Baj, B., Molecular biology of adenosine triphosphate-sensitive potassium channels. Endocr. Rev., 1999, 20, 101–135.

  • Joseph, C., Koster, M. A. P. and Nichols, C. G., Perspectives in diabetes diabetes and insulin secretion. The ATP-sensitive K+ channel (KATP) connection. Diabetes, 2005, 54, 3065–3072.

  • Ashcroft, F. M., ATP-sensitive potassium channelopathies: focus on insulin secretion. J. Clin. Invest., 2005, 115, 2047–2058.

  • Liu, G. J. et al., ATP-sensitive potassium channels induced in liver cells after transfection with insulin cDNA and the GLUT2 transporter regulate glucose-stimulated insulin secretion. FASEB J., 2003, 17, 1682–1684.

  • Ashcroft, F. M., Puljung, M. C. and Vedovato, N., Neonatal diabetes and the KATP channel from mutation to therapy. Trends Endocrinol. Metab., 2017, 28, 377–387.

  • Rolland, J.-F., Henquin, J.-C. and Gilon, P., Feedback control of the ATP-sensitive K+ current by cytosolic Ca2+ contributes to oscillations of the membrane potential in pancreatic cells. Diabetes, 2002, 51, 376–384.

  • Nakano, K., Suga, S., Takeo, T., Ogawa, Y., Suda, T., Kanno, T. and Wakui, M., Intracellular Ca2+ modulation of ATP-sensitive K+ channel activity in acetylcholine-induced activation of rat pancreatic-beta cells. Endocrinology, 2002, 143, 569–576.

  • Kane, G. C. et al., KCNJ11 gene knockout of the Kir6.2 KATP channel causes maladaptive remodeling and heart failure in hypertension. Hum. Mol. Genet., 2006, 15, 2285–2297.

  • Liang, W. et al., The opening of ATP-sensitive K+ channels protects H9c2 cardiac cells against the high glucose-induced injury and inflammation by inhibiting the ROS-TLR4-necroptosis pathway. Cell Physiol. Biochem., 2017, 41, 1020–1034.

  • Choi, S. M. et al., Elevated aldolase 1A, retrogene 1 expression induces cardiac apoptosis in rat experimental autoimmune myocarditis model. Can. J. Physiol. Pharm., 2020, 98, 373–382.

  • Meiboom, S. and Gill, D., Modified spin-echo method for measuring nuclear relaxation times. Rev. Sci. Instrum., 1958, 29, 688–691.

  • Jiang, Chun-Ying, Yang, Kang-Min, Yang, Liu, Miao, Zhao-Xia, Wang, Ying-Hong and Zhu, Hai-Bo, A 1H NMR-based metabonomic investigation of time-related metabolic trajectories of the plasma, urine, and liver extracts of hyperlipidemic hamsters. PLoS ONE, 2013, 8, e66786.

  • Merrifield, C. A. et al., A metabolic system-wide characterization of the pig: a model for human physiology. Mol. BioSyst., 2011, 7, 2577–2588.

  • Chuichi Kawai, M., From myocarditis to cardiomyopathy: mechanisms of inflammation and cell death: learning from the past for the future. Circulation, 1999, 99, 1091–1100.

  • Wu, B. et al., The impact of circulating mitochondrial DNA on cardiomyocyte apoptosis and myocardial injury after TLR4 activation in experimental autoimmune myocarditis. Cell. Physiol. Biochem., 2017, 42, 713–728.

  • Fu, C., Dai, X., Yang, Y., Lin, M., Cai, Y. and Cai, S., Dexmedetomidine attenuates lipopolysaccharide-induced acute lung injury by inhibiting oxidative stress, mitochondrial dysfunction and apoptosis in rats. Mol. Med. Rep., 2017, 15, 131–138.

  • Yucel, G. et al., Lipopolysaccharides induced inflammatory responses and electrophysiological dysfunctions in human-induced pluripotent stem cell derived cardiomyocytes. Sci. Rep., 2017, 7, 2935.

  • Rogatzki, M. J., Ferguson, B. S., Goodwin, M. L. and Gladden, L. B., Lactate is always the end product of glycolysis. Front. Neurosci., 2015, 9, 22.

  • Kawase, T. et al., Validation of lactate level as a predictor of early mortality in acute decompensated heart failure patients who entered intensive care unit. J. Cardiol., 2015, 65, 164–170.

  • Mishra, R., Emancipator, S. N., Kern, T. and Simonson, M. S., High glucose evokes an intrinsic proapoptotic signaling pathway in mesangial cells. Kidney Int., 2005, 67, 82–93.

  • Tinker, A., Aziz, Q. and Thomas, A., The role of ATP-sensitive potassium channels in cellular function and protection in the cardiovascular system. Br. J. Pharmacol., 2014, 171, 12–23.

  • Richer, M. J. and Horwitz, M. S., The innate immune response: an important partner in shaping coxsackievirus-mediated autoimmunity. J. Innate. Immun., 2009, 1, 421–434.

  • Salas, M. A. et al., The signalling pathway of CaMKII-mediated apoptosis and necrosis in the ischemia/reperfusion injury. J. Mol. Cell Cardiol., 2010, 48, 1298–1306.

  • Anderson, M. E., Braun, A. P., Schulman, H. and Premack, B. A., Multifunctional Ca2+/calmodulin-dependent protein kinase mediates Ca(2+)-induced enhancement of the L-type Ca2+ current in rabbit ventricular myocytes. Circ. Res., 1994, 75, 854–861.

  • Mundiña-Weilenmann, C., Vittone, L., Ortale, M., de Cingolani, G. C. and Mattiazzi, A., Immunodetection of phosphorylation sites gives new insights into the mechanisms underlying phospholamban phosphorylation in the intact heart. J. Biol. Chem., 1996, 271, 33561–33567.

  • Maier, L. S. and Bers, D. M., Calcium, calmodulin, and calcium– calmodulin kinase II: Heartbeat to heartbeat and beyond. J. Mol. Cell Cardiol., 2002, 34, 919–939.

  • Ferrero, P. et al., Ca2+/calmodulin kinase II increases ryanodine binding and Ca2+-induced sarcoplasmic reticulum Ca2+ release kinetics during beta-adrenergic stimulation. J. Mol. Cell Cardiol., 2007, 43, 281–291.

  • MacDonnell, S. M. et al., Adrenergic regulation of cardiac contractility does not involve phosphorylation of the cardiac ryanodine receptor at serine 2808. Circ. Res., 2008, 102, e65–e72.

  • Zhang, T. and Brown, J. H., Role of Ca2+/calmodulin-dependent protein kinase II in cardiac hypertrophy and heart failure. Cardiovasc. Res., 2004, 63, 476–486.


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  • KCNJ11 Downregulation Stimulates Cardiac Cell Apoptosis in Myocarditis

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Authors

Seungmin Choi
Department of Life Sciences, Korea University, Seoul 02841, Korea, Republic of
Joo Hee Chung
Korea Basic Science Institute, Seoul Center, Seoul 02841, Korea, Republic of
Myung-Hee Nam
Korea Basic Science Institute, Seoul Center, Seoul 02841, Korea, Republic of
Eunjung Bang
Korea Basic Science Institute, Western Seoul Center, Seoul 03759, Korea, Republic of
Jong Bok Seo
Korea Basic Science Institute, Seoul Center, Seoul 02841, Korea, Republic of
Sung-Gil Chi
Department of Life Sciences, Korea University, Seoul 02841, Korea, Republic of

Abstract


One of the inflammatory heart diseases, viz. acute myocarditis, occurs due to cardiac cell death. However, the molecular mechanism underlying cell death remains largely unexplored. In this study, we report that the down-regulation of KCNJ11, a central subunit of the ATP-sensitive potassium (KATP) channel plays a key role in the reduction of blood glucose, and is involved in apoptosis of cardiac cells. Using proteomics analysis of experimental autoimmune myocarditis (EAM), we show that the KCNJ11 level marked by decreased, whereas Camk2a expression increased significantly in EAM tissues by 16 and 20 days, compared to control tissues. Using 1H-MAS NMR we also show that glucose levels were slightly elevated in EAM tissues. In vitro assays using H9c2 cardiac cells revealed that both lipopolysaccharide (LPS) and high glucose treatment decreased cell viability, in which toxicity was attenuated by treatment with KATP pharmacological openers, but not by the KATP blockers (Gli and 5-HD). Apoptosis induced by LPS or high glucose treatment was suppressed by Ca2+ chelator (BAPTA-AM) treatment. We found that KCNJ11 levels had decreased in cardiac cells by LPS or high glucose treatment, and siRNA-mediated knockdown of KCNJ11 expression further stimulated the LPS- or high glucose-induced apoptosis. Together, our results demonstrate first that KCNJ11 is down-regulated under inflammation and high glucose conditions and its inactivation facilitates cardiac cell apoptosis. We assume that down-regulation of KCNJ11 has an effect on the development of myocarditis.

Keywords


Apoptosis, Blood Glucose Lipopolysaccharide, Cardiac Cells, Myocarditis.

References





DOI: https://doi.org/10.18520/cs%2Fv119%2Fi7%2F1106-1112