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Pleiotropic, non-cell death-associated effects of inhibitors of receptor-interacting protein kinase 1 in the heart

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Abstract

Inhibition of receptor-interacting protein kinase 1 (RIP1) has been recognized as a compelling tool for limiting necroptosis. Recent findings have indicated that RIP1 inhibitor, necrostatin-1 (Nec-1), is also able to modify heart function under non-cell death conditions. In this study, we investigated its underlying molecular mechanisms and compared with those of novel pharmacologically improved agents (Nec-1s and GSK’772) and its inactive analog (Nec-1i). Heart function was examined in Langendorff-perfused rat hearts. Certain proteins regulating myocardial contraction–relaxation cycle and oxidative stress (OS) were evaluated by immunoblotting and as the extent of lipid peroxidation, protein carbonylation and nitration, respectively. In spite of the increase of left ventricular developed pressure (LVDP) due to treatment by both Nec-1 and Nec-1i, only the former agent increased the phosphorylation of Ca2+/calmodulin-dependent protein kinase II delta (CaMKIIδ) at threonine 287 and cardiac myosin-binding protein-C (cMyBPc) at serine 282. In contrast, Nec-1s did not elicit such changes, while it also increased LVDP. GSK’772 activated CaMKIIδ-phospholamban (PLN) axis. Neither protein kinase A (PKA) nor its selected molecular targets, such as serine 16 phosphorylated PLN and sarco/endoplasmic reticulum Ca2+-ATPase 2a (SERCA2a), were affected by either RIP1 inhibitor. Nec-1, like other necrostatins (Nec-1i, Nec-1s), but not GSK’772, elevated protein tyrosine nitration without affecting other markers of OS. In conclusion, this study indicated for the first time that Nec-1 may affect basal heart function by the modulation of OS and activation of some proteins of contraction–relaxation cycle.

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Fig. 1
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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

BP:

Blood pressure

CaMKIIδ:

Ca2+/calmodulin-dependent protein kinase II delta

cMYBPc:

Cardiac myosin-binding protein-C

DNPH:

2,4-Dinitrophenylhydrazine

ECC:

Excitation–contraction coupling

EG:

Electrogram

GSK’772:

GSK2982772

HR:

Heart rate

IDO:

Indoleamine 2,3-dioxygenase

LVDP:

Left ventricular developed pressure

Nec-1:

Necrostatin-1

Nec-1i:

Necrostatin-1i

Nec-1s:

Necrostatin-1s

OS:

Oxidative stress

PKA:

Protein kinase A

PLN:

Phospholamban

PVDF:

Polyvinylidene difluoride

RIP1:

Receptor-interacting protein kinase 1

TBARS:

Thiobarbituric acid reactive substances

SERCA2a:

Sarco/endoplasmic reticulum Ca2+-ATPase

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Acknowledgements

The authors would like to thank Ms. J. Formankova for her skillful technical assistance and Mgr. I. Jarabicova for her assistance with biochemical methods.

Funding

This study was supported by The Slovak Research and Development Agency, Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic (APVV‐15‐607, APVV-20-0242, APVV-19-0540, VEGA SR 1/0016/20 and 2/0141/18).

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Comenius University in Bratislava, Odbojarov 10, 832 32, Bratislava, Slovak Republic

    C. Horvath, A. Szobi & A. Adameova

  2. Centre of Experimental Medicine, Institute for Heart Research, Slovak Academy of Sciences, Bratislava, Slovak Republic

    L. Kindernay, T. Ravingerova & A. Adameova

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  1. C. Horvath
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  2. A. Szobi
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Correspondence to A. Adameova.

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The authors declare that they have no conflict of interest.

Ethical approval

Protocol of this study has been approved by the Ethics Committee of the Faculty of Pharmacy, Comenius University in Bratislava. All procedures described herein were performed in accordance with the Guide for the care and Use of Laboratory Animals, published by the US National Institutes of Health (Guide, NRC 2011) and approved by the Animal Health and Welfare Division of the State Veterinary and Food Administration of the Slovak Republic.

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Horvath, C., Szobi, A., Kindernay, L. et al. Pleiotropic, non-cell death-associated effects of inhibitors of receptor-interacting protein kinase 1 in the heart. Mol Cell Biochem 476, 3079–3087 (2021). https://doi.org/10.1007/s11010-021-04136-y

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