Kivonat:
Embryonic stem cell (ESC)-derived cardiomyocytes are a promising
cell source for the screening for potential cytoprotective
molecules against ischemia/reperfusion injury, however, little
is known on their behavior in hypoxia/reoxygenation conditions.
Here we tested the cytoprotective effect of the NO-donor SNAP
and its downstream cellular pathway. Mouse ESC-derived
cardiomyocytes were subjected to 150-min simulated ischemia (SI)
followed by 120-min reoxygenation or corresponding non-ischemic
conditions. The following treatments were applied during SI or
normoxia: the NO-donor S-Nitroso-N-acetyl-D,L-penicillamine
(SNAP), the protein kinase G (PKG) inhibitor, the KATP channel
blocker glibenclamide, the particulate guanylate cyclase
activator brain type natriuretic peptide (BNP), and a non-
specific NO synthase inhibitor (N-Nitro-L-arginine, L-NNA) alone
or in different combinations. Viability of cells was assayed by
propidium iodide staining. SNAP attenuated SI-induced cell death
in a concentration-dependent manner, and this protection was
attenuated by inhibition of either PKG or KATP channels.
However, SI-induced cell death was not affected by BNP or by L-
NNA. We conclude that SNAP protects mESC-derived cardiomyocytes
against SI/R injury and that soluble guanylate-cyclase, PKG, and
KATP channels play a role in the downstream pathway of SNAP-
induced cytoprotection. The present mESC-derived cardiomyocyte-
based screening platform is a useful tool for discovery of
cytoprotective molecules.