Ca(2+)-regulatory proteins in cardiomyocytes from the right ventricle in children with congenital heart disease

BACKGROUND: Hypoxia and hypertrophy are the most frequent pathophysiological consequence of congenital heart disease (CHD) which can induce the alteration of Ca(2+)-regulatory proteins and inhibit cardiac contractility. Few studies have been performed to examine Ca(2+)-regulatory proteins in human cardiomyocytes from the hypertrophic right ventricle with or without hypoxia. METHODS: Right ventricle tissues were collected from children with tetralogy of Fallot [n = 25, hypoxia and hypertrophy group (HH group)], pulmonary stenosis [n = 25, hypertrophy group (H group)], or small isolated ventricular septal defect [n = 25, control group (C group)] during open-heart surgery. Paraffin sections of tissues were stained with 3,3′-dioctadecyloxacarbocyanine perchlorate to measure cardiomyocyte size. Expression levels of Ca(2+)-regulatory proteins [sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2a), ryanodine receptor (RyR2), sodiumcalcium exchanger (NCX), sarcolipin (SLN) and phospholamban (PLN)] were analysed by means of real-time PCR, western blot, or immunofluorescence. Additionally, phosphorylation level of RyR and PLN and activity of protein phosphatase (PP1) were evaluated using western blot. RESULTS: Mild cardiomyocyte hypertrophy of the right ventricle in H and HH groups was confirmed by comparing cardiomyocyte size. A significant reduction of SERCA2a in mRNA (P<0.01) was observed in the HH group compared with the C group. The level of Ser(16)-phosphorylated PLN was down-regulated (P<0.01) and PP1 was increased (P<0.01) in the HH group compared to that in the C group. CONCLUSIONS: The decreased SERCA2a mRNA may be a biomarker of the pathological process in the early stage of cyanotic CHD with the hypertrophic right ventricle. A combination of hypoxia and hypertrophy can induce the adverse effect of PLN-Ser(16) dephosphorylation. Increased PP1 could result in the decreased PLN-Ser(16) and inhibition of PP1 is a potential therapeutic target for heart dysfunction in pediatrics.