Chapter 1 The Anti-Apoptotic Effect of Transgenic Akt1 Gene on Cultured New-Born Rats Cardiomyocytes Mediated by Ultrasound/Microbubbles Destruction

摘要

The purpose of this study was to transfect exogenous Akt1 to 293FT cells and cultured new-born rats cardiomyocytes respectively for exploring the feasibility and safety of the new method. In addition, the protective effects on cultured cardiomyocytes suffering from ischemia/reperfusion injury (I/R inury) of the transgenic Akt1 gene mediated by ultrasound-induced microbubble destruction (US/MB) were also observed. 293FT cells and cardiomyocytes were divided into four groups, group A: only pEGFPC1- Akt1; group B: MB + pEGFPC1-Akt1; group C: US + pEGFPC1-Akt1; group D: US + MB + pEGFPC1-Akt1. The results showed that microbubble alone could not deliver exogenous genes to target cells without ultrasound, but the latter could without the help of microbubble, ultrasound with simultaneous microbubble could enhance the transfection rate significantly. Ultrasound frequency 1.7 MHz, mechanical index (MI) 1.5, irradiation time 2 min, MB volume concentration 15 %. According to the above ultrasound parameters, Akt1 gene was transfected to cultured cardiomyocytes and then allowed them to suffer from I/R injury, our results showed that the transgenic Akt1 gene was expressed and had significant anti-apoptotic effect. The results suggested that US/MB was a promising gene delivery system for gene therapy in heart diseases.