دورية أكاديمية
[Construction of VEGF recombinant plasmid pcDNA/V and its expression in model rats with acute myocardial ischemia].
| العنوان: | [Construction of VEGF recombinant plasmid pcDNA/V and its expression in model rats with acute myocardial ischemia]. |
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| المؤلفون: | Wang YM; Experiment Center, Capital University of Medical Sciences, Beijing 100054, China., Liu B, Sun LC, Yan YD, Si Y, Qi YH |
| المصدر: | Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2006 Mar; Vol. 22 (2), pp. 220-5. |
| نوع المنشور: | English Abstract; Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | Chinese |
| بيانات الدورية: | Publisher: Ke xue chu ban she Country of Publication: China NLM ID: 9426463 Publication Model: Print Cited Medium: Print ISSN: 1000-3061 (Print) Linking ISSN: 10003061 NLM ISO Abbreviation: Sheng Wu Gong Cheng Xue Bao Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Beijing : Ke xue chu ban she, |
| مواضيع طبية MeSH: | Genetic Therapy*, Myocardial Infarction/*therapy , Recombinant Proteins/*biosynthesis , Recombinant Proteins/*therapeutic use , Vascular Endothelial Growth Factor A/*genetics, Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane/blood supply ; Disease Models, Animal ; Humans ; Male ; Myocardial Infarction/metabolism ; Myocardial Infarction/pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/genetics ; Transfection ; Vascular Endothelial Growth Factor A/biosynthesis |
| مستخلص: | The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF. |
| المشرفين على المادة: | 0 (Recombinant Proteins) 0 (VEGFA protein, human) 0 (Vascular Endothelial Growth Factor A) |
| تواريخ الأحداث: | Date Created: 20060413 Date Completed: 20090723 Latest Revision: 20121115 |
| رمز التحديث: | 20240628 |
| PMID: | 16607947 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1000-3061 |
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