Effects of phospholamban antisense RNA on SR Ca~(2+)-ATPase and [Ca~(2+)]i in rat cardiomyocytes by adeno-associated virus vector / 中国病理生理杂志

ABSTRACT

AIM: To investigate the effect of phospholamban antisense RNA (asPLB) on the activity of sarco-endoplasmic reticulum (SR) Ca 2+-ATPase, and the change of intracellular free Ca 2+ concentration ([Ca 2+]i) in rat cardiomyocytes by adeno-associated virus(AAV) vector. METHODS: rAAV-asPLB and rAAV-LacZ were constructed by AAV Helper-Free System. RT-PCR and Western blotting were used to determine the mRNA and protein expression of PLB. The activity of SR Ca 2+-ATPase and the [Ca 2+]i were measured. RESULTS: Compared to controls, the PLB mRNA and protein expression reduced in rat cardiomyocytes transfected with rAAV-asPLB. The activity of Ca 2+-ATPase was increased. In rest state, the level of [Ca 2+]i in rAAV-asPLB transfected group was decreased. The level of [Ca 2+]i was increased when induced by isoproterenol. CONCLUSION: rAAV-asPLB vector disrupts the expression of PLB, enhances the activity of Ca 2+-ATPase, reduces the resting [Ca 2+]i and enhances the isoproterenol-induced [Ca 2+]i.