中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (15): 2785-2788.doi: 10.3969/j.issn.1673-8225.2012.15.027

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

经心包腔途径转染提高大鼠体内心肌细胞转染的效率*★

齐迎春，谢晓华，陈  雯，李朝晖   

  1. 解放军总医院南楼综合外一科，北京市  100853
  • 收稿日期:2012-01-30 修回日期:2012-03-02 出版日期:2012-04-08 发布日期:2012-04-08
  • 通讯作者: 谢晓华，硕士，主任医师，教授，博士生导师，解放军总医院南楼综合外一科，北京市 100853 n4xxh@126.com
  • 作者简介:齐迎春★，女，1978年生，辽宁省凌源市人，汉族，2001年华北煤炭医学院毕业，硕士，医师，主要从事高血压心脏病的机制研究。shepherdessqi@hotmail.com
  • 基金资助:

    军队“十五”医药卫生科研基金重点课题(02Z0010)。

Improvement of transfection efficiency with plasmid transfected to the myocardium of rats intrapericardially in vivo

Qi Ying-chun, Xie Xiao-hua, Chen Wen, Li Zhao-hui   

  1. First Department of Integrated Surgery, South Building, General Hospital of Chinese PLA, Beijing  100853, China
  • Received:2012-01-30 Revised:2012-03-02 Online:2012-04-08 Published:2012-04-08
  • Contact: Xie Xiao-hua, Master, Chief physician, Professor, Doctoral supervisor, First Department of Integrated Surgery, South Building, General Hospital of Chinese PLA, Beijing 100853, China n4xxh@126.com
  • About author:Qi Ying-chun★, Master, Physician, First Department of Integrated Surgery, South Building, General Hospital of Chinese PLA, Beijing 100853, China shepherdessqi@hotmail.com
  • Supported by:

    the Military Medical Science Foundation during the Tenth Five-year Period, No. 02Z0010*

PDF

279

摘要：

背景：如何获得安全、有效、广泛心肌组织的转染一直是国内外学者研究的热点。
目的：探讨能够改善动物水平心肌组织非病毒载体的转染效率、增强基因导入靶向性的方法。
方法：以β半乳糖苷酶质粒PLacZ作为报告基因，将Wistar大鼠随机分为5组：①空白组。②心包腔内组：心包腔注射质粒+微泡+酶混悬液，超声导入。③心包腔内阴性对照组：以生理盐水代替质粒干预。④舌下静脉组：舌下静脉注射质粒+微泡混悬液，超声导入。⑤舌下静脉阴性对照组：以生理盐水代替质粒干预。注射6 d后处死，进行心、肺、肝、肾组织的X-gal染色。
结果与结论：转染6 d后，仅有心包腔内组大鼠的部分心肌细胞在心尖、心室及心房水平可见明显的蓝染，其他各组大鼠心肌X-gal染色为阴性；各组大鼠肺、肝、肾组织X-gal染色均为阴性。提示采用心包腔内途径转染、再辅以超声微泡导入以及酶类的使用，可明显改善质粒对在体心肌细胞的转染效率，且不伴有心外组织目的基因的表达，具有较好的靶向性。
关键词：心包；心肌细胞；siRNA；RNAi；转染；基因
doi:10.3969/j.issn.1673-8225.2012.15.027

关键词: 心包, 心肌细胞, siRNA, RNAi, 转染, 基因

Abstract:

BACKGROUND: It has always been the highlights to find how to accomplish safe, effective and extensive transfection of the myocardium.
OBJECTIVE: To explore a new feasible method for improving transfection efficiency of non-viral plasmids to the myocardium and for obtaining satisfactory myocardial targeting.
METHODS: We chose PLacZ as a report gene. Wistar rats were randomly divided into five groups: control group, intrapericardial group, intrapericardial negative group, sublingual vein group, sublingual vein negative group. For all the subsets of experiments, hearts were harvested at 6 days after injection. Tissues of heart, lung, liver and kidney were stained with X-gal, and PlacZ gene expression of the heart and other non-targeted organs was observed to decide the gene transfection status and myocardial targeting.
RESULTS AND CONCLUSION: After 6 days of transfection, only rats in the intrapericardial group were stained blue in parts of the myocardium on the atrial, ventricular and apex level. Other groups were negative for X-gal staining in the myocardium, lung, liver and kidney. The intrapericardial injection of microbubbles and some enzymes, as well as plasmids, with the aid of ultrasound, can improve transfection efficiency of plasmids significantly and the heart targeting is satisfactory.
Qi YC, Xie XH, Chen W, Li ZH. Improvement of transfection efficiency with plasmid transfected to the myocardium of rats intrapericardially in vivo. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(15): 2785-2788.    
[http://www.crter.cn  http://en.zglckf.com]
 

中图分类号: