Overexpression of sarcoplasmic reticulum calcium ATPase in isolated rat myocytes mediated by recombinant adenovirus

Objective: To assess value of SERCA2a in the gene therapy of congestive heart failure, furthermore to analysis the effect of transfeetion to pathophysiology. Methods: A recombinant adenovirus, named rAd-trSERCA2a, was constructed with AdEasy system, in which the SERCA2a cDNA was inserted under the control of CMV promoter.The cardiac myoeytes of newborn rat were cultured, and were transfected with rAd-trSERCA2a.RTPCR and Western Blotting were performed to detect the expression of SERCA2a in cardiac myoeytes. Results: the sequence of SERCA2a was stably existed in rAd-trSERCA2a, which was tested by PCR and southern blot.The quantity of mRNA of SERCA2a in cardiac myoeytes increased 3.6 times compared with the control, and the protein level is 1.5 times of the control, 72 hours after the transfeetion. Conclusion: This recombinant virus, rAd-trSERCA2a, can effectively delivery the SERCA2a gene into cultured cardiac myoeytes and may became a feasible tool to modulate myoeyte Ca²⁺ homeostasis in the failure heart.