PCMV_4-hTGFβ_1在大鼠深Ⅱ度烫伤创面愈合及瘢痕形成中的作用  

Effects of PCMV_4-hTGFβ_1 as nucleic acid vaccine on Ⅱ°burn wound healing and postburn scarring in rats

作　　者：贾赤宇[1] 陈壁[1] 王德文[2] 

机构地区：[1]第四军医大学西京医院烧伤科,西安710032 [2]北京解放军军事医学科学院基础所二所八室

出　　处：《中华整形烧伤外科杂志》1999年第3期170-172,共3页

摘　　要：目的观察与人类巨细胞病毒载体结合的转化生长因子β_1(PCMV_4-hTGFβ_1)重组质粒裸 DNA 直接注射于深Ⅱ度烫伤大鼠体内后的基因表达规律及对创面愈合的作用,探讨其作为核酸疫苗的可行性,为烧伤的基因治疗提供理论基础和实验依据。方法大鼠随机分为烫伤+核酸疫苗组;单纯核酸疫苗组;对照组。采用 ELISA 方法检测裸 DNA 注射后血清中中和抗体水平;Southern 杂交、原位杂交检测裸 DNA 注射后在肌肉中存留时间,组织特染法检测组织中Ⅰ/Ⅲ型胶原的变化。结果裸 DNA 肌注7天后血清中可检测出抗 TGFβ_1中和抗体,21天达其峰值,28天有轻度下降。Southern 杂交显示裸 DNA 注射后,5min 即可检测出高浓度 TGFβ_1,1h 后浓度开始下降,3h 后仍可检测出。原位杂交在肌注5天后,肌肉中还有阳性反应。核酸疫苗组创面愈合速度在前9天较对照组快,差异有显著性意义(P<0.05),在后期与对照组间差异无显著性意义(P>0.05),但Ⅰ/Ⅲ型胶原比例明显下降(P<0.01)。结论 PCMV_4-hTGFβ_1即可加速创面愈合,又可控制Ⅰ型胶原合成,控制瘢痕增生。证实了其作为核酸疫苗治疗烧伤创面及控制瘢痕形成是可行的。Objective To investigate the gene expression of PCMV_4-hTGFβ_1 as nucleic acid vac- cine regulation and the effects on burn wound healing and scarring in rats and study the feasibility of gene therapy in burns.Methods Sixty wistar rats were divided into three groups:①burn+nucleic acid vac- cine;②nucleic acid vaccine only(control);③burn only(control).ELISA was used to determine the dy- namic changes in anti-TGFβ_1 neutralizing antibody in serum;determination of the quantities of DNA re- tained in muscles by Southern blot and in situ hybridization;observation of the ratio in collagen Ⅰ/Ⅲ during the wound healing by special stain method.Results The level of anti-TGFβ_1 antibody in serum reached the peak at the third week after naked DNA was injected,and a slight drop in the 4th week. TGFβ_1 plasmid DNA could be detected 5 minute after injection,and lasted 3 hours.In situ hybridization showed a positive staining in muscle fibers 5 days after injection.During the day 0—day 9,the wound healing speed in vaccine group was faster than control,and after day 10,no significent difference was found between groups,but the ratio of collagen Ⅰ/Ⅲ was reduced remarkbly in vaccine group.Conclu- sion Injection of PCMV_4-hTGFβ_1 in rats can really cause general immune response reaction.It was showed that PCMV_4-hTGFβ_1 was similar to TGFβ_1,and it bad the effect of stimulating of epidermic cells to accelerate wound healing at early stage,and at later stage,it was similar to anti-TGFβ_1 neutralizing an- tibody,having the effect of inhibiting hyperplasia of scar.So it is confirmed that PCMV_4-hTGFβ_1 as nu- cleic acid vaccine has the effect of promoting healing of burn wound and controling the formation of scar.

关 键 词：创面愈合 烧伤 TGFΒ1 核酸疫苗 瘢痕愈合 

分 类 号：R644[医药卫生—外科学]