Recombinant plasmid pIRES2-EGFP-HCN2 improved pacing function in canine model of sick sinus syndrome / 中华心血管病杂志
Chinese Journal of Cardiology
; (12): 1126-1130, 2006.
Article
em Zh
| WPRIM
| ID: wpr-238471
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct plasmid expressing pacemaker gene pIRES2-EGFP-HCN2 and study its effects in transfected atrial myocytes in vitro and in canine model of sick sinus syndrome (SSS).</p><p><b>METHODS</b>mHCN2 gene was isolated from PTR plasmids and cloned into eukaryotic expression plasmid pIRES2-EGFP. Recombinant plasmids pIRES2-EGFP-HCN2 was transfected with by electroporation into neonatal atrial cardiomyocytes or injected to the sinoatrial (SA) region of canines with SSS induced by catheter and chemical ablation. pIRES2-EGFP-HCN2 expression was detected under fluorescence microscope and confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Spontaneous beating rate in atrial cardiomyocytes was detected with light microscope.</p><p><b>RESULTS</b>EGFP expression was seen in transfected atrial cardiomyocytes 24 to 48 hours after transfection and the spontaneous beating rate was significantly increased than that in non-transfected atrial cardiomyocytes [(180 +/- 11) bpm vs (140 +/- 14) bpm, P < 0.05]. Heart rate was significantly increased 24 hours post recombinant plasmids pIRES2-EGFP-HCN2 injection compared to saline injection in canines with SSS [(150 +/- 13) bpm vs (105 +/- 17) bpm, P < 0.05]. Green fluorescence was also detected in frozen SA tissue sections of canines injected with recombinant plasmids pIRES2-EGFP-HCN2 and the production amplified by RT-PCR was about 300 bp which is consistent with mHCN2 gene fragment.</p><p><b>CONCLUSION</b>The recombinant eukaryotic expression plasmid pIRES2-EGFP-HCN2 can improve pacing function in atrial myocytes and in canine model of SSS.</p>
Texto completo:
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Base de dados:
WPRIM
Assunto principal:
Plasmídeos
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Síndrome do Nó Sinusal
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Terapêutica
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Técnicas In Vitro
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Terapia Genética
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Expressão Gênica
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Técnicas de Transferência de Genes
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Miócitos Cardíacos
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Modelos Animais de Doenças
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Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Cardiology
Ano de publicação:
2006
Tipo de documento:
Article