中华超声影像学杂志
中華超聲影像學雜誌
중화초성영상학잡지
CHINESE JOURNAL OF ULTRASONOGRAPHY
2011年
4期
351-354
,共4页
王兴华%雷成功%乔英艳%石颖%王芹秀
王興華%雷成功%喬英豔%石穎%王芹秀
왕흥화%뢰성공%교영염%석영%왕근수
超声检查%微气泡%转染
超聲檢查%微氣泡%轉染
초성검사%미기포%전염
Ultrasonography%Microbubbles%Transfection
目的 探讨细胞膜孔开放及含氟烷气体白蛋白外膜在超声微泡介导GFP转染C57810及mdx小鼠骨骼肌细胞中的机制.方法 以肌细胞膜缺损为主要病理改变的mdx小鼠与正常C57810小鼠为研究对象,目的基因GFP与Optison或SonoVue混合注入小鼠胫前肌,一侧胫前肌经超声辐照,另一侧胫前肌不经超声辐照.C57810小鼠作为正常对照,实验分组如下:①C57810小鼠生理盐水组(4条左胫前肌);②C57810小鼠生理盐水+超声组(4条右胫前肌);③C57810小鼠Optison组(4条左胫前肌);④C57810小鼠Oprison+超声组(4条右胫前肌);⑤C57810小鼠SonoVue组(4条左胫前肌);⑥C57810小鼠SonoVue+超声组(4条右胫前肌).mdx肌营养不良小鼠实验分组如下:①mdx小鼠生理盐水组(4条左胫前肌);②mdx小鼠生理盐水+超声组(4条右胫前肌);③mdx小鼠+Optison组(4条左胫前肌);④mdx小鼠Optison+超声组(4条右胫前肌);⑤mdx小鼠SonoVue组(4条左胫前肌);⑥mdx小鼠SonoVue+超声组(4条右胫前肌).1周后处死小鼠,荧光显微镜观察发出绿色荧光者为GFP阳性肌纤维细胞,计数最大GFP阳性肌纤维细胞数,作为GFP基因转染效率指标.结果 正常C57810小鼠:①无超声作用时,与阴性对照组比较,Optison微泡显著提高GFP基因转染水平(P<0.01),SonoVue微泡不提高GFP基因转染水平;②有超声作用时,与阴性对照组比较,Optison微泡显著提高GFP基因转染水平(P<0.01);③有超声作用时,与阴性对照组比较,SonoVue微泡显著提高GFP基因转染水平(P<0.01).mdx小鼠:①与正常C57810小鼠比较,GFP单独(生理盐水组)显著提高GFP基因转染水平(P<0.01),Optison微泡显著提高GFP基因转染水平(P<0.01),SonoVue微泡显著提高GFP基因转染水平(P<0.01);②与阴性对照组比较,Optison微泡显著提高GFP基因转染水平(P<0.01),SonoVue微泡显著提高GFP基因转染水平(P<0.01).结论 细胞膜孔开放是微泡提高基因转染水平的重要因素,含氟烷气体白蛋白外膜是Optison微泡提高GFP转染水平的主要成分.
目的 探討細胞膜孔開放及含氟烷氣體白蛋白外膜在超聲微泡介導GFP轉染C57810及mdx小鼠骨骼肌細胞中的機製.方法 以肌細胞膜缺損為主要病理改變的mdx小鼠與正常C57810小鼠為研究對象,目的基因GFP與Optison或SonoVue混閤註入小鼠脛前肌,一側脛前肌經超聲輻照,另一側脛前肌不經超聲輻照.C57810小鼠作為正常對照,實驗分組如下:①C57810小鼠生理鹽水組(4條左脛前肌);②C57810小鼠生理鹽水+超聲組(4條右脛前肌);③C57810小鼠Optison組(4條左脛前肌);④C57810小鼠Oprison+超聲組(4條右脛前肌);⑤C57810小鼠SonoVue組(4條左脛前肌);⑥C57810小鼠SonoVue+超聲組(4條右脛前肌).mdx肌營養不良小鼠實驗分組如下:①mdx小鼠生理鹽水組(4條左脛前肌);②mdx小鼠生理鹽水+超聲組(4條右脛前肌);③mdx小鼠+Optison組(4條左脛前肌);④mdx小鼠Optison+超聲組(4條右脛前肌);⑤mdx小鼠SonoVue組(4條左脛前肌);⑥mdx小鼠SonoVue+超聲組(4條右脛前肌).1週後處死小鼠,熒光顯微鏡觀察髮齣綠色熒光者為GFP暘性肌纖維細胞,計數最大GFP暘性肌纖維細胞數,作為GFP基因轉染效率指標.結果 正常C57810小鼠:①無超聲作用時,與陰性對照組比較,Optison微泡顯著提高GFP基因轉染水平(P<0.01),SonoVue微泡不提高GFP基因轉染水平;②有超聲作用時,與陰性對照組比較,Optison微泡顯著提高GFP基因轉染水平(P<0.01);③有超聲作用時,與陰性對照組比較,SonoVue微泡顯著提高GFP基因轉染水平(P<0.01).mdx小鼠:①與正常C57810小鼠比較,GFP單獨(生理鹽水組)顯著提高GFP基因轉染水平(P<0.01),Optison微泡顯著提高GFP基因轉染水平(P<0.01),SonoVue微泡顯著提高GFP基因轉染水平(P<0.01);②與陰性對照組比較,Optison微泡顯著提高GFP基因轉染水平(P<0.01),SonoVue微泡顯著提高GFP基因轉染水平(P<0.01).結論 細胞膜孔開放是微泡提高基因轉染水平的重要因素,含氟烷氣體白蛋白外膜是Optison微泡提高GFP轉染水平的主要成分.
목적 탐토세포막공개방급함불완기체백단백외막재초성미포개도GFP전염C57810급mdx소서골격기세포중적궤제.방법 이기세포막결손위주요병리개변적mdx소서여정상C57810소서위연구대상,목적기인GFP여Optison혹SonoVue혼합주입소서경전기,일측경전기경초성복조,령일측경전기불경초성복조.C57810소서작위정상대조,실험분조여하:①C57810소서생리염수조(4조좌경전기);②C57810소서생리염수+초성조(4조우경전기);③C57810소서Optison조(4조좌경전기);④C57810소서Oprison+초성조(4조우경전기);⑤C57810소서SonoVue조(4조좌경전기);⑥C57810소서SonoVue+초성조(4조우경전기).mdx기영양불량소서실험분조여하:①mdx소서생리염수조(4조좌경전기);②mdx소서생리염수+초성조(4조우경전기);③mdx소서+Optison조(4조좌경전기);④mdx소서Optison+초성조(4조우경전기);⑤mdx소서SonoVue조(4조좌경전기);⑥mdx소서SonoVue+초성조(4조우경전기).1주후처사소서,형광현미경관찰발출록색형광자위GFP양성기섬유세포,계수최대GFP양성기섬유세포수,작위GFP기인전염효솔지표.결과 정상C57810소서:①무초성작용시,여음성대조조비교,Optison미포현저제고GFP기인전염수평(P<0.01),SonoVue미포불제고GFP기인전염수평;②유초성작용시,여음성대조조비교,Optison미포현저제고GFP기인전염수평(P<0.01);③유초성작용시,여음성대조조비교,SonoVue미포현저제고GFP기인전염수평(P<0.01).mdx소서:①여정상C57810소서비교,GFP단독(생리염수조)현저제고GFP기인전염수평(P<0.01),Optison미포현저제고GFP기인전염수평(P<0.01),SonoVue미포현저제고GFP기인전염수평(P<0.01);②여음성대조조비교,Optison미포현저제고GFP기인전염수평(P<0.01),SonoVue미포현저제고GFP기인전염수평(P<0.01).결론 세포막공개방시미포제고기인전염수평적중요인소,함불완기체백단백외막시Optison미포제고GFP전염수평적주요성분.
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 control:①C57B10 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 groups:① 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 mice:①Optison 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(P<O.01).Mdx mice:①Compared 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.
