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Cardioprotective Effects of Beta3-Adrenergic Receptor (β3-AR) Pre-, Per-, and Post-treatment in Ischemia–Reperfusion

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Abstract

The β3-AR (beta3-adrenergic receptor) is resistant to short-term agonist-promoted desensitization and delivers a constant intracellular signal, making this receptor a potential target in acute myocardial infarction (AMI).

Aim

To investigate whether selective modulation of β3-AR prior to or during ischemia and/or reperfusion may be cardioprotective.

Methods

Isolated perfused rat hearts were exposed to 35-min regional ischemia (RI) and 60-min reperfusion. The β3-AR agonist (BRL37344, 1 μM) or antagonist (SR59230A, 0.1 μM) was applied: (i) before RI (PreT) or (ii) last 10 min of RI (PerT) or (iii) onset of reperfusion (PostT) or (iv) during both PerT+PostT. Nitric oxide (NO) involvement was assessed, using the NOS inhibitor, L-NAME (50 μM). Endpoints were functional recovery, infarct size (IS), cGMP levels, and Western blot analysis of eNOS, ERKp44/p42, PKB/Akt, and glycogen synthase kinase-3β (GSK-3β).

Results

Selective treatment with BRL significantly reduced IS. L-NAME abolished BRL-mediated cardioprotection. BRL (PreT) and BRL (PerT) significantly increased cGMP levels (which were reduced by L-NAME) and PKB/Akt phosphorylation. BRL (PostT) produced significantly increased cGMP levels, PKB/Akt, and ERKp44/p42 phosphorylation. BRL (PerT+PostT) caused significant eNOS, PKB/Akt, ERKp44/p42, and GSK-3β phosphorylation.

Conclusion

β3-AR activation by BRL37344 induced significant cardioprotection regardless of the experimental protocol. However, the pattern of intracellular signaling with each BRL treatment differed to some degree and suggests the involvement of cGMP, eNOS, ERK, GSK-3β, and particularly PKB/Akt activation. The data also suggest that clinical application of β3-AR stimulation should preferably be incorporated during late ischemia or/and early reperfusion.

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Authors and Affiliations

  1. Biomedical Research and Innovation Platform, South African Medical Research Council, Building D, Medicina, Francie van Zijl Drive, Parow Valley, Cape Town, Western Cape, South Africa

    Ruduwaan Salie

  2. Faculty of Medicine and Health Sciences, Division of Medical Physiology, University of Stellenbosch, PO Box 19063, Cape Town, South Africa

    Ruduwaan Salie, Aisha Khlani Hassan Alsalhin, Erna Marais & Amanda Lochner

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  1. Ruduwaan Salie
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  2. Aisha Khlani Hassan Alsalhin
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Correspondence to Ruduwaan Salie.

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Experimental Animals

Male Wistar rats (230 to 250 g) were used in this study. The handling of laboratory animals was in accordance with the ethical guidelines as set out by the University of Stellenbosch, Faculty of Medicine and Health Sciences Ethics Committee and the South African National Standard for Care and Use of Animals for Scientific Purpose (SANS 10386: 2008). The rats had free access to food and water prior to experimentation. Rats were anesthetized with sodium pentobarbital (30 mg/rat) by intraperitoneal injection before removal of hearts.

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Salie, R., Alsalhin, A.K.H., Marais, E. et al. Cardioprotective Effects of Beta3-Adrenergic Receptor (β3-AR) Pre-, Per-, and Post-treatment in Ischemia–Reperfusion. Cardiovasc Drugs Ther 33, 163–177 (2019). https://doi.org/10.1007/s10557-019-06861-5

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