Rationale:
The β
2
-adrenoceptor (β
2
-AR), a prototypical G protein-coupled receptor (GPCR), couples to both G
s
and G
i
proteins. Stimulation of theβ
2
-AR is beneficial to humans and animals with heart failure presumably because it activates the downstream G
i
-PI3K-Akt cell survival pathway. Cardiac β
2
-AR signaling can be regulated by crosstalk or heterodimerization with other GPCRs, but the physiological and pathophysiological significance of this type of regulation has not been sufficiently demonstrated.
Objective:
Here, we aim to investigate the potential cardioprotective effect of β
2
-adrenergic stimulation with a subtype-selective agonist, (R,R')-4-methoxy-1-naphthylfenoterol (MNF), and to decipher the underlying mechanism with a particular emphasis on the role of heterodimerization of β
2
-ARs with another GPCR, 5-hydroxytryptamine receptors 2B (5-HT
2B
Rs).
Methods and Results:
Using pharmacological, genetic and biophysical protein-protein interaction approaches, we studied the cardioprotective effect of the β
2
-agonist, MNF, and explored the underlying mechanism in both in vivo in mice and cultured rodent cardiomyocytes insulted with doxorubicin (Dox), hydrogen peroxide (H
2
O
2
) or ischemia/reperfusion. In Dox-treated mice, MNF reduced mortality and body weight loss, while improving cardiac function and cardiomyocyte viability. MNF also alleviated myocardial ischemia/reperfusion injury. In cultured rodent cardiomyocytes, MNF inhibited DNA damage and cell death caused by Dox, H
2
O
2
or hypoxia/reoxygenation. Mechanistically, we found that MNF or another β
2
-agonist zinterol markedly promoted heterodimerization of β
2
-ARs with 5-HT
2B
Rs. Upregulation of the heterodimerized 5-HT
2B
Rs and β
2
-ARs enhanced β
2
-AR-stimulated Gi-Akt signaling and cardioprotection while knockdown or pharmacological inhibition of the 5-HT
attenuated β
2
-AR-stimulated G
i
signaling and cardioprotection.
Conclusions:
These data demonstrate that the β
2
-AR-stimulated cardioprotective Gi signaling depends on the heterodimerization of β
2
-ARs and 5-HT
2B
Rs.