Ultrasonic destruction of albumin microbubbles enhances gene transfection and expression in cardiac myocytes  被引量:2

Ultrasonic destruction of albumin microbubbles enhances gene transfection and expression in cardiac myocytes

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作  者:WANG Guo-zhong LIU Jing-hua LUE Shu-zheng LUE Yun GUO Cheng-jun ZHAO Dong-hui FANG Dong-ping HE Dong-fang ZHOU Yuan GE Chang-jiang 

机构地区:[1]Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Vessel Diseases, Beijing 100029, China

出  处:《Chinese Medical Journal》2011年第9期1395-1400,共6页中华医学杂志(英文版)

摘  要:Background It has been proven that ultrasonic destruction of microbubbles can enhance gene transfection efficiency into the noncardiac cells, but there are few reports about cardiac myocytes. Moreover, the exact mechanisms are not yet clear; whether the characteristic of microbubbles can affect the gene transfection efficiency or not is still controversial.This study was designed to investigate whether the ultrasound destruction of gene-loaded microbubbles could enhance the plasmids carried reporter gene transfection in primary cultured myocardial cell, and evaluate the effects of microbubbles characteristics on the transgene expression in cardiac myocytes.Methods The β-galactosidase plasmids attached to the two types of microbubbles, air-contained sonicated dextrose albumin (ASDA) and perfluoropropane-exposed sonicated dextrose albumin (PESDA) were prepared. The gene transfection into cardiac myocytes was performed in vitro by naked plasmids, ultrasound exposure, ultrasonic destruction of gene-loaded microbubbles and calcium phosphate precipitation, and then the gene expression and cell viability were analyzed.Results The ultrasonic destruction of gene-loaded microbubbles enhanced gene expression in cardiac myocytes compared with naked plasmid transfection ((51.95±2..41) U/g or (29.28±3.65) U/g vs. (0.84-0.21) U/g, P <0.01), and ultrasonic destruction PESDA resulted in more significant gene expression than ASDA ((51.95e2.41) U/g vs. (29.28±3.65)U/g, P <0.05). Ultrasonic destruction of microbubbles during calcium phosphate precipitation gene transfection enhanced 3-galactosidase activity nearly 8-fold compared with calcium phosphate precipitation gene transfection alone ((111.35±11.21) U/g protein vs. (14.13±2.58) U/g protein, P<0.01). Even 6 hours after calcium phosphate precipitation gene transfection, ultrasound-mediated microbubbles destruction resulted in more intense gene expression ((35.63±7.65)U/g vs. (14.13±2.58) U/g, P<0.05 ).CoBackground It has been proven that ultrasonic destruction of microbubbles can enhance gene transfection efficiency into the noncardiac cells, but there are few reports about cardiac myocytes. Moreover, the exact mechanisms are not yet clear; whether the characteristic of microbubbles can affect the gene transfection efficiency or not is still controversial.This study was designed to investigate whether the ultrasound destruction of gene-loaded microbubbles could enhance the plasmids carried reporter gene transfection in primary cultured myocardial cell, and evaluate the effects of microbubbles characteristics on the transgene expression in cardiac myocytes.Methods The β-galactosidase plasmids attached to the two types of microbubbles, air-contained sonicated dextrose albumin (ASDA) and perfluoropropane-exposed sonicated dextrose albumin (PESDA) were prepared. The gene transfection into cardiac myocytes was performed in vitro by naked plasmids, ultrasound exposure, ultrasonic destruction of gene-loaded microbubbles and calcium phosphate precipitation, and then the gene expression and cell viability were analyzed.Results The ultrasonic destruction of gene-loaded microbubbles enhanced gene expression in cardiac myocytes compared with naked plasmid transfection ((51.95±2..41) U/g or (29.28±3.65) U/g vs. (0.84-0.21) U/g, P <0.01), and ultrasonic destruction PESDA resulted in more significant gene expression than ASDA ((51.95e2.41) U/g vs. (29.28±3.65)U/g, P <0.05). Ultrasonic destruction of microbubbles during calcium phosphate precipitation gene transfection enhanced 3-galactosidase activity nearly 8-fold compared with calcium phosphate precipitation gene transfection alone ((111.35±11.21) U/g protein vs. (14.13±2.58) U/g protein, P<0.01). Even 6 hours after calcium phosphate precipitation gene transfection, ultrasound-mediated microbubbles destruction resulted in more intense gene expression ((35.63±7.65)U/g vs. (14.13±2.58) U/g, P<0.05 ).Co

关 键 词:ultrasound MICROBUBBLES gene transfer cardiac myocytes 

分 类 号:Q78[生物学—分子生物学] Q463

 

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