Attenuation of cardiac ischemia-reperfusion injury by sodium thiosulfate is partially dependent on the effect of cystathione beta synthase in the myocardium
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Our early studies have shown that sodium thiosulfate (STS) treatment attenuated the ischemia-reperfusion (IR)-induced injury in an isolated rat heart model by decreasing apoptosis, oxidative stress, and preserving mitochondrial function. Hydrogen sulfide, the precursor molecule is reported to have similar efficacy. This study aims to investigate the role of endogenous hydrogen sulfide in STS-mediated cardioprotection against IR in an isolated rat heart model. d, l-propargylglycine (PAG), an inhibitor of cystathionine γ-lyase was used as endogenous H2S blocker. In addition, we used the hypoxia-reoxygenation (HR) model to study the impact of STS in cardiomyocytes (H9C2) and fibroblast (3T3) cells. STS treatment to animal and cells prior to IR or HR decreased cell injury. The sensitivity of H9C2 and 3T3 cells towards HR (6 h hypoxia followed by 12 h reoxygenation) challenge varies, where, 3T3 cells exhibited higher cell death (54%). Cells treated with PAG prior to STS abrogate the protective effect in 3T3 (cell viability 61%) but not in H9C2 (cell viability 82%). Further evaluation in rat heart model showed partial recovery (46% RPP) of heart from those hearts pretreated with PAG prior to STS condition. In conclusion, we demonstrated that STS-mediated cardioprotection to IR-challenged rat heart is not fully dependent on endogenous H2S level and this dependency may be linked to higher fibroblast content in rat heart.
The authors would like to acknowledge the Indian Council for Medical Research (ICMR), Government of India, New Delhi, for supporting this research through grant-in-aid (No. 5/4/1-14/12-NCD-II).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The study protocol involving experimentation on animals was approved by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India, with prior approval of the Institutional Animal Ethical Committee (IAEC, SASTRA University, No.: 229/SASTRA/IAEC/RPP).
- 4.Neri, M., & Riezzo, I. (2017). Ischemia/reperfusion injury following acute myocardial infarction: a critical issue for clinicians and forensic pathologists. Mediators of Inflammation, 2017, 7018393.Google Scholar
- 6.Ravindran, S., Boovarahan, S. R., Shanmugam, K., Vedarathinam, R. C., & Kurian, G. A. (2017). Sodium thiosulfate preconditioning ameliorates ischemia/reperfusion injury in rat hearts via reduction of oxidative stress and apoptosis. Cardiovascular Drugs and Therapy, 31, 511–524.CrossRefGoogle Scholar
- 11.Feoktistova, M., Geserick, P., & Leverkus, M. (2016) Crystal violet assay for determining viability of cultured cells. Cold Spring Harbour Protocols. 2016, pdb.prot087379. https://doi.org/10.1101/pdb.prot087379.
- 12.Kasibhatla, S., Amarante-Mendes, G. P., Finucane, D., Brunner, T., Bossy-Wetzel, E., & Green, D. R. (2006). Acridine orange/ethidium bromide (AO/EB) staining to detect apoptosis. CSH Protocols, 2006, 2006.Google Scholar
- 14.Tanzer, M. L., & Gilvarg, C. (1959). Creatine and creatine kinase measurement. Journal of Biological Chemistry, 234, 3201–3204.Google Scholar
- 15.Ytrehus, K., Liu, Y., Tsuchida, A., Miura, T., Liu, G. S., & Yang, X. M., et al. (1994). Rat and rabbit heart infarction: effects of anesthesia, perfusate, risk zone, and method of infarct sizing. American Journal of Physiology, 267, H2383–H2390.Google Scholar
- 16.Sharma, R., Singh Rathore, S., Sharma, P., & Sharma, A. (2009). Estimation of thiosulphate using sodium nitroprusside by a newer photochemical method. Journal of Chemical and Pharmaceutical Research, 1, 321–328.Google Scholar
- 17.Ang, A. D., Konigstorfer, A., Giles, G. I., & Bhatia, M. (2012). Measuring free tissue sulfide. Advances in Biological Chemistry, 02No.04, 6.Google Scholar
- 18.Palmer, J. W., Tandler, B., & Hoppel, C. L. (1977). Biochemical properties of subsarcolemmal and interfibrillar mitochondria isolated from rat cardiac muscle. Journal of Biological Chemistry, 252, 8731–8739.Google Scholar
- 25.Frazier, A. E., & Thorburn, D. R. (2012). Biochemical Analyses of the Electron Transport Chain Complexes by Spectrophotometry. In: Wong, L. J. (ed.), Mitochondrial Disorders. Methods in Molecular Biology (Methods and Protocols), Vol 837. Humana Press.Google Scholar
- 27.Lee, C.-Y., Hwang, J.-H., Lee, Y.-S., & Cho, K.-S. (1995). Purification and characterization of mouse liver rhodanese. Journal of Biochemistry and Molecular Biology, 28, 170–176.Google Scholar