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兔心肌内注射人心房钠尿肽基因裸质粒的生物活性和安全性(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2004年第2期
页码:
121-123
栏目:
基础研究
出版日期:
2004-03-01

文章信息/Info

Title:
Bioactivity and safety of direct intramyocardial injection of naked plasmid DNA encoding human atrial natriuretic peptide
作者:
黄国明; 李庚山; 朱刚艳; 赖亦木; 张宏祥; 王晶
武汉大学第一临床学院心血管内科,湖北 武汉 430060
Author(s):
HUANG Guo-ming; LI Geng-shan; ZHU Gang-yan; LAI Yi-mu; ZHANG Hong-xiang; WANG Jing
Department of Cardiology, People's Hospital of Wuhan University, Wuhan, Hubei 430060, China
关键词:
心房钠尿肽; 质粒; 心肌; 基因表达; 兔
Keywords:
atrial natriuretic peptide;  plasmid;  myocardium;  gene expression;  rabbit
分类号:
R542.2
DOI:
-
文献标识码:
A
摘要:
目的:评价心肌内注射人心房钠尿肽(hANF)基因裸质粒(pCR3.hANF)的生物活性和安全性。方法:家兔20只开胸后用微量注射器将800μg的pCR3.hANF(实验组,n=16)或pCR3(对照组,n=4)平均注射至左室前壁的四个部位;通过逆转录-聚合酶链式反应(RT-PCR)、Southern杂交和放射免疫测定(RIA),检测hANF基因的表达。结果:实验组hANF基因在心肌中的表达显著高于对照组(P<0.01),且在第30天达到高峰;肝、肺、肾组织中无hANF基因表达(均P>0.05);兔心肌mRNA的表达与hANF的表达显著相关(r=0.819,P<0.01)。结论:hANF基因裸质粒心肌内注射是一安全、有效的转导途径。
Abstract:
AIM:To evaluate the safety and bioactivity of direct intramyocardial injection of naked DNA encoded human atrial natriuretic peptide(hANF). METHODS: hANF gene injection was performed in 20 rabbits. For each rabbit, 800μg of reconstructed naked plasmid DNA encoded hANF-pCR3. hANF (n=16) or pCR3 (n=4,control) in 80μl normal saline was injected averagely into four sites on the anterior wall of the left ventricular via a micro-injector. The expressions of hANF mRNA in left ventricular myocardium,liver,lung,and kidney were identified by reverse transcription polymerase chain reaction followed by Southern blot analysis. Plasma ANF levels were determined by radioimmunoassay (RIA). RESULTS: The naked pCR3. hANF caused a significant expression of hANF mRNA inventricular myocardium and amarked increase in plasma ANF levels 10,20,30,40 days after the gene delivery (all P<0.01). No hANF mRNA was detected indistal tissues, including liver,lung,and kidney (all P>0.05). CONCLUSION: Direct intramyocardial injection of naked plasmid DNA encoded hANF is a safe and bioactive method for gene delivery.

参考文献/References

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[2]金冬雁,黎孟枫,等译.《分子克隆实验指南》[M].第2版,北京:科学出版社,1992.

[3]Losordo DW,Vale PR,Symes JF,et al.Gene therapy for myocardial angiogenesis initial clinical results with direct myocardial injection of phVEGF165 as sole therapy for myocardialischemia[J]. Circulation, 1998, 98:2800-2804.

[4]Schwarz E,Speakman MT,Patterson M,et al.Evaluation of the effects of intramycardial injection of DNA expressing vascular endothelial growth factor in a myocardial infarction modelinthe rat-angiogenesis and angiomaformation[J]. J Am CollCardiol,2000,35:1323-1326.

备注/Memo

备注/Memo:
收稿日期：2003-6-2
更新日期/Last Update: 2004-03-01