Oxysterol, 5α-cholestan-3-one, modulates a contractile response to β2-adrenoceptor stimulation in the mouse atria: Involvement of NO signaling

- We study changes in responses to β2-adrenergic agonist fenoterol at the mouse atria.
- 5α-Cholestan-3-one decreases inotropy of fenoterol despite increased Ca2 + transient.
- This occurs due to upregulation of Gi-protein-Akt-NO-protein kinase G pathway.
- The enhancement of NO-signaling depends on membrane cholesterol content.
- The oxysterol is a potential modulator of the β2-adrenergic signaling in the atria.

AimAtrial β2-adrenoceptors provide an important mechanism for regulation of cardiac function and changes in their downstream signaling are involved in processes underlying heart disorders. We have investigated the mechanism by which the cholesterol metabolite 5α-cholestan-3-one (5ɑCh3) modulates inotropic effect of β2-adrenoceptor agonist fenoterol.Main methodsAtria from mice were electrically stimulated and changes in contraction amplitude in response to fenoterol were studied in 5ɑCh3-pretreated samples. Intracellular Ca2 + and NO levels were estimated using fluorescent dyes Fluo-4 and DAF-FM, respectively.Key findingsBy itself 5αCh3 that appears in the circulation under some pathological conditions had a negligible influence on contraction, Ca2 +-transient and NO production. However, pretreatment with 5αCh3 markedly attenuated the positive inotropic effect of fenoterol which was accompanied by an increase in the NO synthesis. Unexpectedly, the oxysterol also augmented an enhancement of Ca2 +-transient amplitude in response to fenoterol. Under conditions of a pharmacological inhibition of Gi-protein/Akt/NO synthase/protein kinase G signaling, 5αCh3 augmented the inotropic effect of fenoterol. Herein, Akt antagonist suppressed the increase in NO production, while inhibition of NO synthesis did not modify the increased amplitude of the Ca2 +-transient. Along similar lines, enrichment of plasma membranes with cholesterol reduced the stimulatory effect of 5αCh3 on β2-adrenoceptor-evoked NO production, but not on the Ca2 +-transient amplitude, leading to an elevation of the positive inotropic response to fenoterol.SignificanceThese data suggest that 5ɑCh3 potentiates the effect of pharmacological β2-adrenoceptor activation on both NO production and Ca2 + transient via independent mechanisms, thereby affecting the positive inotropy.

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