The experiment in the mechanisms of microbubble-mediated GFP gene enhancement in skeletal muscle in C57B10/mdx mice / 中华超声影像学杂志

ABSTRACT

Objective To investigate the role of sonoporation and the deblic of microbubbles with perfluoropropane gas and albumin in the mechanisms of microbubble-mediated gene enhancement by experimenting in skeletal muscle in C57B10/mdx mice. Methods Plasmid DNA (10 μg) encoding green fluorescent protein (GFP) was mixed with Optison or SonoVue dissolved in saline and injected into the tibialis anterior (TA) muscle of /C57B10/mdx mice with and without adjunct ultrasound. The efficiencies of GFP transgene expression were determined under different experimental conditions. C57B10 mice as normal control①C57B10 mice + saline (4 left TAs);②C57B10 mice + saline + ultrasound (4 right TAs) ;③C57B10 mice + Optison(4 left TAs);④C57B10 mice+ Optison + ultrasound(4 right TAs);⑤ C57B10 mice + SonoVue(4 left TAs) ;⑥C57B10 mice + SonoVue + ultrasound(4 right TAs). Mdx mice groups① mdx mice + saline(4 left TAs) ;② mdx mice + saline + ultrasound(4 right TAs);③ mdx mice + Optison (4 left TAs) ; ④ mdx mice + Optison + ultrasound (4 right TAs); ⑤mdx mice + SonoVue(4 left TAs) ;⑥mdx mice + SonoVue + ultrasound(4 right TAs). Mice were sacrificed 1 week after plasmid DNA injection. Fibres with fluorescence green signals were determined as GFP-positive fibres by fluorescence microscopy. Readout was performed on the section with the maximum number of transfected fibers. Results C57B10 mice ?Optison without ultrasound had significantly increased gene expression compared with negative control ( P <0. 01). SonoVue without ultrasound did not enhance gene expression. ?Optison with ultrasound had significantly increased gene expression compared with negative control (P < 0.01). ?SonoVue with ultrasound had significantly increased gene expression compared with negative control ( P<0. 01).Mdx mice? Compared with C57B10 mice, GFP alone demonstrated significant GFP expression in mdx mice ( P <0. 01) , Optison demonstrated significant GFP expression in mdx mice ( P <0.01), and SonoVue demonstrated significant GFP expression in mdx mice ( P <0. 01). ?Microbubble groups (Optison and SonoVue) had significantly increased gene expression compared with negative control (P <0. 01). Conclusions In the mechanisms of microbubble-mediated gene enhancement, sonoporation is the key step. The deblic of microbubbles with perfluoropropane gas and albumin is the main constituent in the mechanisms of Optison-mediated gene enhancement. fibers.Results C5781 0 mice①Optison without ultrasound had significantly increased gene expressioncompared with negative control(P<0.01).SonoVue without ultrasound did not enhance gene expression.②Optison with ultrasound had significantly increased gene expression compared with negative control(P<0.01).③SonoVue with ultrasound had significantly increased gene expression compared with negativecontr01(PMdx mice①Compared with C57810 mice,GFP alone demonstrated significant GFPexpression in mdx mice(P<0.01),0prison demonstrated significant GFP expression in mdx mice(P<0.01),and SonoVue demonstrated significant GFP expression in mdx mice(P<0.01).②Microbubblegroups(0ptison and SonoVue)had significantly increased gene expression compared with negative control(P<0.01).Conclusions In the mechanisms of microbubble-mediated gene enhancement,sonoporation isthe key step.The deblic of microbubbles with perfluoropropane gas and albumin is the main constituent inthe mechanisms of Optison-mediated gene enhancement.