The Effect of Androgen Deprivation on the Expression of Connexin-43 mRNA in the Heart

Document Type : Research Articles

Authors

• Mahnaz Ghowsi ¹
• Nazli Khajehnasiri ²
• Sajjad Sisakhtnezhad ¹

¹ Department of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran

² Department of Biological Sciences, Faculty of Basic Sciences, Higher Education Institute of Rab-Rashid, Tabriz, Iran.

https://doi.org/10.22067/jcmr.2021.71395.1010

Abstract

Connexin-43 (Cx-43) plays axial roles in the propagation of action potentials and contractile coupling in heart. Down-regulation of Cx-43 in heart is associated with arrhythmia, dilated cardiomyopathy and heart failure. To date, no studies have examined the effects of androgen deprivation therapy (ADT)-induced hypogonadism on the expression of Cx-43 in heart. This study investigated the effects of testosterone deprivation and its replacement with testosterone on the expression of Cx-43 mRNA and muscle-specific miRNAs miR-206 and miR-1, as two potential regulators of the Cx-43 protein expression in the ventricular tissue. Accordingly, 21 male Wistar rats were divided into three groups: Ι) Normal control, П) ORX-S: castrated rats serving as animal models for ADT and receiving the sesame oil as a solvent of testosterone enanthate for 10 weeks, and Ш) ORX-T: these animals were castrated, receiving testosterone enanthate (25 mg/kg) for 10 weeks. The relative expression of Cx-43 mRNA, miR-206 and miR-1 was determined by qRT-PCR. Cx-43 mRNA was found to be decreased in the ORX-S group. The Cx-43 mRNA was up-regulated after the administration of testosterone enanthate. There were no significant changes in miR-206 and miR-1 levels in the ORX-S and ORX-T groups when compared to the controls. Our results indicated that testosterone should be regarded as an important factor in the regulation of the Cx-43 mRNA expression in heart, and testosterone deprivation may down-regulate the Cx-43 mRNA expression; however, it doesn’t alter miR-1 and miR-206 levels. These results suggest that ADT-induced hypogonadism may put males at risk for cardiac dysfunctions.

Keywords

• Connexin-43
• miR-206
• miR-1
• Heart
• Testosterone
• Androgen deprivation therapy

20.1001.1.20089147.2021.13.1.8.1

References

Anderson C., Catoe H. and Werner R. (2006) MIR-206 regulates connexin43 expression during skeletal muscle development. Nucleic Acids Research 34: 5863-5871.

Care A., Catalucci D., Felicetti F., Bonci D., Addario A., Gallo P., et al. (2007) MicroRNA-133 controls cardiac hypertrophy. Nature Medicine 13: 613-618.

Chang K.-T., Cheng C.-F., King P.-C., Liu S.-Y. and Wang G.-S. (2017) CELF1 mediates connexin 43 mRNA degradation in dilated cardiomyopathy. Circulation Research 121: 1140-1152.

Curcio A., Torella D., Iaconetti C., Pasceri E., Sabatino J., Sorrentino S., et al. (2013) MicroRNA-1 downregulation increases connexin 43 displacement and induces ventricular tachyarrhythmias in rodent hypertrophic hearts. PloS One 8: e70158.

Dong S., Cheng Y., Yang J., Li J., Liu X., Wang X., et al. (2009) MicroRNA expression signature and the role of microRNA-21 in the early phase of acute myocardial infarction. Journal of Biological Chemistry 284: 29514-29525.

Lian D., Xiong X., Liu Y. and Wang J. (2014) miRNA-1: functional roles and dysregulation in heart disease. Molecular BioSystems 10: 2775-2782.

Firestone G. L. and Kapadia B.J. (2012) Minireview: regulation of gap junction dynamics by nuclear hormone receptors and their ligands. Molecular Endocrinology 26: 1798-1807.

Foley PL. (2005) common surgical procedures in rodents', laboratory animal medicine and management. office of animal research education and compliance, University of Virginia, Charlottesville, VA, USA: International Veterinary Information Service, Ithaca NY (www. ivis. org).

Huang C.-K., Lee S. O., Chang E., Pang H. and Chang C. (2016) Androgen receptor (AR) in cardiovascular diseases. The Journal of Endocrinology 229: R1.

Huang Y. M., Li W. W., Wu J., Han M. and Li B. H. (2019) The diagnostic value of circulating microRNAs in heart failure. Experimental and Therapeutic Medicine 17: 1985-2003.

Jansen J. A., van Veen T. A., de Bakker J. M. and van Rijen H. V. (2010) Cardiac connexins and impulse propagation. Journal of Molecular and Cellular Cardiology 48: 76-82.

Kamal D. A. M., Ibrahim S. F. and Mokhtar M. H. (2020) Effects of testosterone on the expression of Connexin 26 and Connexin 43 in the uterus of rats during early pregnancy. In Vivo 34: 1863-1870.

Keating N. L., O'Malley A. J. and Smith M. R. (2006) Diabetes and cardiovascular disease during androgen deprivation therapy for prostate cancer. Journal of clinical oncology 24: 4448-4456.

Kramer M. F. (2011) Stem-loop RT-qPCR for miRNAs. Current Protocols in Molecular Biology. 95: 15.10. 1-15.10. 15.

Kura B., Kalocayova B., Devaux Y. and Bartekova M. (2020) Potential clinical implications of miR-1 and miR-21 in heart disease and cardioprotection. International Journal of Molecular Sciences 21: 700.

Kurtenbach S., Kurtenbach S. and Zoidl G. (2014) Gap junction modulation and its implications for heart function. Frontiers in Physiology 5: 82.

Liu N. and Olson E. N. (2010) MicroRNA regulatory networks in cardiovascular development. Developmental Cell 18: 510-525.

Malkin C.J. and Channer K. (2009) Testosterone in chronic heart failure. Advances in the Management of Testosterone Deficiency 37: 183-196.

Manzano-Fernández S., Pastor-Pérez F. J., Barquero-Pérez Ó, Pascual-Figal DA, Goya-Esteban R, Rojo-Álvarez J. L., et al. (2011) Short-term variability of heart rate turbulence in chronic heart failure. Journal of Cardiac Failure 17: 735-741.

McCarthy J. J. (2011) The MyomiR network in skeletal muscle plasticity. Exercise and Sport Sciences Reviews 39: 150.

Michela P., Velia V., Aldo P. and Ada P. (2015) Role of connexin 43 in cardiovascular diseases. European Journal of Pharmacology 768: 71-76.

Mitchelson K. R. and Qin W.-Y. (2015) Roles of the canonical myomiRs miR-1,-133 and-206 in cell development and disease. World Journal of Biological Chemistry 6: 162.

Mitsuzuka K. and Arai Y. (2018) Metabolic changes in patients with prostate cancer during androgen deprivation therapy. International Journal of Urology 25: 45-53.

Nielsen S., Hvid T., Kelly M., Lindegaard B., Dethlefsen C., Winding K., et al. (2014) Muscle specific miRNAs are induced by testosterone and independently upregulated by age. Frontiers in Physiology 4: 394.

Oyamada M., Takebe K. and Oyamada Y. (2013) Regulation of connexin expression by transcription factors and epigenetic mechanisms. Biochimica et Biophysica Acta (BBA)-Biomembranes 1828: 118-133.

Pongkan W., Chattipakorn S. C. and Chattipakorn N. (2015) Chronic testosterone replacement exerts cardioprotection against cardiac ischemia-reperfusion injury by attenuating mitochondrial dysfunction in testosterone-deprived rats. PloS One 10: e0122503.

Schmittgen T. D. and Livak K.J. (2008) Analyzing real-time PCR data by the comparative CT method. Nature Protocols 3: 1101-1108.

Shahani S., Braga-Basaria M. and Basaria S. (2008) Androgen deprivation therapy in prostate cancer and metabolic risk for atherosclerosis. The Journal of Clinical Endocrinology & Metabolism 93: 2042-2049.

Shan Z.-X., Lin Q.-X., Fu Y.-H., Deng C.-Y., Zhou Z.-L., Zhu. J.-N., et al. (2009) Upregulated expression of miR-1/miR-206 in a rat model of myocardial infarction. Biochemical and Biophysical Research Communications 381: 597-601.

Wieciech I, Grzesiak M., Knapczyk-Stwora K., Pytlik A. and Słomczyńska M. (2014) Influence of the antiandrogen flutamide on connexin 43 (Cx43) gene and protein expression in the porcine placenta and uterus during pregnancy. Folia Biologica (Kraków) 62: 367-375.

Wu C.-H., Yang J.-G., Yang J-J., Lin Y.-M., Tsai H.-D., Lin C.-Y., et al. (2010) Androgen excess down-regulates connexin43 in a human granulosa cell line. Fertility and Sterility 94: 2938-2941.

Xu M., Hu C., Khan H.-h., Shi F.-h., Cong X.-d., Li Q., et al. (2016) Argirein alleviates stress-induced and diabetic hypogonadism in rats via normalizing testis endothelin receptor A and connexin 43. Acta Pharmacologica Sinica 37: 246-254.

Yan Y., Dang H., Zhang X., Wang, X., and Liu, X. (2020) The protective role of MiR-206 in regulating cardiomyocytes apoptosis induced by ischemic injury by targeting PTP1B. Bioscience Reports 40: BSR20191000.

Yang B., Lin H., Xiao J., Lu Y., Luo X., Li B., et al. (2007) The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. Nature Medicine 13: 486-491.

Zhang J., Jin S., Zhao J. and Li H. (2016) Effect of dibutyl phthalate on expression of connexin 43 and testosterone production of leydig cells in adult rats. Environmental Toxicology and Pharmacology 47: 131-135.