Inhibition of phosphodiesterases leads to prevention of the mitochondrial permeability transition pore opening and reperfusion injury in cardiac H9c2 cells.

Abstract:

PURPOSE:We tested if inhibition of phosphodiesterases (PDEs) with IBMX (1-methyl-3-isobutylxanthine) can modulate the mitochondrial permeability transition pore (mPTP) opening by inactivating glycogen synthase kinase 3β (GSK-3β). METHODS:H9c2 cells were exposed to 600 μM H(2)O(2) for 20 min to cause the mPTP opening. Mitochondrial membrane potential (ΔΨm) was assessed by imaging cells loaded with tetramethylrhodamine ethyl ester (TMRE). Cell viability was measured with propidium iodide (PI) fluorometry using a fluorescence reader. Ischemia/reperfusion injury was induced by exposing cells to ischemic solution for 90 min followed by 30 min of reperfusion. RESULTS:IBMX reduced loss of ΔΨm caused by H(2)O(2), indicating that inhibition of PDEs can prevent the mPTP opening. However, IBMX could not inhibit the pore opening in cells transfected with the constitutively active GSK-3β (GSK-3β-S9A) mutant, suggesting a critical role of GSK-3β in the action of IBMX. IBMX also reduced reperfusion injury in a GSK-3β dependent manner. In support, IBMX increased GSK-3β phosphorylation at Ser(9), an effect that was reversed by both the PKA inhibitor H89 and the PKG inhibitor KT5823. In support, IBMX activated both PKA and PKG. IBMX failed to prevent the loss of ΔΨm in the presence of H89 or PKA siRNA. Similarly, both KT5823 and PKG siRNA reversed the protective effect of IBMX. CONCLUSION:Inhibition of PDEs prevents the mPTP opening by inactivating GSK-3β through PKA and PKG. GSK-3β is a common downstream target of PKA and PKG. Inhibition of PDEs may be a useful approach to prevent reperfusion injury.

journal_name

Cardiovasc Drugs Ther

authors

Chanoit G,Zhou J,Lee S,McIntosh R,Shen X,Zvara DA,Xu Z

doi

10.1007/s10557-011-6310-z

subject

Has Abstract

pub_date

2011-08-01 00:00:00

pages

299-306

issue

4

eissn

0920-3206

issn

1573-7241

journal_volume

25

pub_type

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