Trasferring EGFP gene into skeletal muscle of rats mediated by microbubbles

Objective To explore the feasibility of enhanced green fluorescent protein(EGFP)transfected into skeletal muscle of rats under the condition of the increasing in vascular permeability.Methods Twenty Sprague-Dawley(SD)rats were randomly divided into four groups,i.e.plasmid group(P),plasmid+ultrasound group(P+U),plasmid+microbub- ble(P+M)and plasmid+ultrasound+microbubble group(P+U+M).Microbubbles were attached with the naked plasmid DNA of EGFP and followed the way of ultrasound-mediated microbubbles destruction to trasfect gene into skeletal muscle of rats.Seven days after gene transfer,the EGFP expression in the muscle was observed under the fluorescence microscope. Results EGFP expression was not observed in the rats of the P groups nor P+M groups,though a small amount of faint green fluorescence was seen in P+U group.Visibility of apparent specificity EGFP expression was observed in P+U+M group,and the fluorescence intensity of P+U+M group was 10 times higer than that of P and P+M groups,but 3 times than that of P+U group(P0.05).The transfection rate of P+U+M group was(42.72±10.07)%,signifiantly higher than that of P+U group(13.62±6.17)%,(P0.05).Conclusion The increasing of capillary permeability caused by de- struction of ultrasound-mediated microbubble contrast agent is one of the major ways for intravascular gene penetrating endo- metrial barriers successfully in spinotrapezius muscle of rats.