Abstract: Objective To construct Ad-hBNP and observe the distribution and the expression of hBNP gene in CHF rats. The study could provide a reference of basic research for BNP in gene therapy. Methords The target gene hBNP were obstained by reverse transcription polymerase chain reaction from human heart. The pAd-hBNP were constructed through AdEasy system. Recombinant plasmid pAd-hBNP were packaged and amplified in 293T cells. Ad-hBNP used in gene therapy were acquired. 24 CHF rats were randomly divided into 4 groups for detecting the serum levels of hBNP with ELISA, the expression of hBNP with RT-PCR and immunohistochemistry in different groups. Results Ad-hBNP were constructed successfully and the titer of purified Ad-hBNP was 4.4275×1012VP/ml. Recombinant adenoviruses were showed as polyhedron shape under electron microscopy. The expression of hBNP gene were detected in CHF rats by intraperitoneal injection Ad-hBNP and it continued at least 7 days. Conclusions Ad-hBNP were constructed successfully by using AdEasy system. Exogenous hBNP were effectively expressed in CHF rats. Compared with the half life of BNP, the prosses that at least continued 7 days were significantly extended.

Key words: Brain natriuretic peptide, Adnovirus, Chronic heart failure, Gene therapy