VEGF_(165)基因对大鼠软组织缺损修复及Ⅰ、Ⅲ型胶原mRNA表达的影响  

The effect of pcDNA3.1-VEGF_(165) recombined vector on wound healing and the expression of collagen type Ⅰ、Ⅲ mRNA in wounded tissue

作　　者：赵冬梅[1] 杨加峰[1] 邱丽萍[1] 武士清[1] 刘军莉[1] 蔡景龙[1] 

机构地区：[1]山东大学第二医院,济南250033

出　　处：《中华整形外科杂志》2006年第6期461-464,共4页Chinese Journal of Plastic Surgery

基　　金：山东省科技攻关计划资助项目(032050112);山东省卫生厅项目(2003)

摘　　要：目的将pcDNA3·1-VEGF165质粒用于软组织缺损局部,观察其对软组织修复的作用和I型、III型胶原mRNA表达的影响。方法SD大鼠32只于两侧臀部制备直径12mm圆形全层皮肤、肌肉的复合软组织缺损;随机分为实验组,对侧为对照组。于实验组注射0·2ml(含200ng)质粒液,对照组以同量生理盐水代替。分别于术后3、5、7、14、30d照相计算创面收缩率、取材。以RT-PCR法检测局部I型、III型胶原mRNA表达;制备常规切片观察局部组织血管形成(MVD)及修复情况。结果所有动物术后无死亡、感染。实验组及对照组创面愈合时间分别为14·2d及17·4d,于1、2周时新生血管密度分别为63·38±9·20、52·72±7·06(P<0·05)及76·64±12·27、66·84±9·82(P<0·05),差别有统计学意义。RT-PCR示I型及III型胶原mRNA于术后3d即开始有表达,2周达到高峰后减弱,实验组表达量较大。结论pcDNA3·1-VEGF165质粒用于创面后,具有上调I型及III型胶原mRNA表达,促进软组织缺损区血管生成、加速创面愈合作用。Objective To examine the effect of pcDNA3.1-VEGF165 vector to the angiogoiesis, expression of collagen type Ⅰ and type Ⅲ mRNA in soft tissue injury model. Methods Thirty two Sprague- Daulay rats,weighted （180 ± 20）g, were made tissue injury in the bilateral of vertebral region. Round wound （diameter 12 mm） was made by perforex on the back, removed the skin and 2 mm muscle, one side was experimental group by random and the other as control. The wound was done with sodium chloride（0.2 ml） in the control group, with the recombinant VEGF165 vector（0.2 ml,200 mg） in the experimental group. The wound healing and other general state of health was observed after the operation. The specimens were obtained at 3,5,7,14 and 30 days after injury . Draw the materials from the rats at the same time, all samples were divided into two parts, one （ 〉 0.1 g）was conserved in refrigerator at - 80℃ , which was extracted total RNA by TRIZOL, design the primer of rat＇ s collagen type Ⅰ and type Ⅲ , RT-PCR analysis indicated that collagen type Ⅰ , Ⅲ . The other was fixed by 10% formalin. Examine wound healing of local tissue and count it＇ s MVD by HE staining. Results All the rabbits were well alive, no death or infection. Wound healing time was shorter than the control one（ 14.2,17.4 d）. Inflammatory cell infiltrate, cellula intersitialis, fibroblast, collagen and the density of angiogenesis were more in the experimental group than in the control one. The MVD was significant difference between the two groups at 1,2 weeks are 63.38 ± 9.20,52.72 ±7.06 and 76.64 ±12.27,66.84 ±9.82（ P 〈 0.05） . The expression of collagen type Ⅰ ,Ⅲ mRNA was found in the third day, the peak was in the second week and then degression. The collagen type Ⅰ ,Ⅲ mRNA and β-actin specificitic belt were found and its initial template volume different, the results was trend of RT-PCR obtained. Conclusions The local application of pcDNA3.1-VEGFIas can enhance the expression of collagen type Ⅰ ,Ⅲ mRNA,e

关 键 词：基因转染 血管内皮生长因子 RT—PCR 基因治疗 

分 类 号：R641[医药卫生—外科学]