0336 : Intracardiac injection of siRNA with temporary aortic ligation is an efficient method to target transient gene expression modulation in the rat left ventricle - 05/05/16
Résumé |
Introduction |
The modulation of gene expression is a widely used strategy to study complex pathological mechanisms. Knock out and transgenic models express their modified genome for their entire life span. Our team developed a surgical procedure in the adult rat to modulate protein expression specifically in the heart adapted from the anterograde coronary injection performed in human patients with a catheter: the intracardiac injection with temporary aortic ligation. For this study, cyclophilin B (CypB) was targeted using small interfering RNA (siRNA)
Material and methods anesthetized |
10 weeks-old Sprague-Dawley rats underwent thoracotomy and 150μg/kg of either NaCl 0.9% (Sham), nontargeting (NT) or CypB siRNA were injected in the left ventricular (LV) cavity after aortic ligation maintained for 20 seconds to allow the coronary diffusion of the siRNA. Three days after the surgery, rats underwent an echography and hearts were harvested to study LV CypB expression by western-blotting.
Results |
Three days after the surgery, rats of every group did not show any alteration of their general condition. In CypB group, CypB expression was decreased by 75% (p<0.05) in LV compared to the Sham group. NT group did not show any alteration of CypB expression. Cardiac echography revealed no difference in the anterior wall thickness (Sham: 1.76±0.04; NT: 1.80±0.05; CypB: 1.75±0.11mm), ejection fraction (Sham: 83.8±0.9; NT: 81.8±1.6; CypB: 78.5±1.7%) and diastolic function measured by early-to-late filing ratio (Sham: 1.19±0.03; NT: 1.13±0.05; CypB: 1.20±0.05).
Conclusion |
Our study showed that intracardiac injection of siRNA with temporary aortic ligation is an efficient method to silence a specific gene, without major post-operative cardiovascular effects. We are undergoing to determine if the CypB silencing is similar in the different cardiac cavities. This technique will allow us to silence or overexpress a protein of interest in vivo.
The author hereby declares no conflict of interest
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Vol 8 - N° 3
P. 257 - avril 2016 Retour au numéro
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