阻断转化生长因子-β信号转导抑制伤口瘢痕的实验研究  

Inhibiting scar formation in rat cutaneous wounds by blocking TGF-β signaling

作　　者：刘伟[1] 蔡泽浩[1] 武晓莉[1] 王丹茹[1] 殷德民[1] 崔磊[1] 曹谊林[1] Michael T.Longaker 

机构地区：[1]上海第二医科大学附属第九人民医院整复外科,200011 [2]Department of urgery,stanford University

出　　处：《中华医学杂志》2003年第1期31-36,共6页National Medical Journal of China

基　　金：上海市科委重大课题研究资助项目 ( 0 0JC14 0 3 2 )

摘　　要：目的　探讨用基因治疗手段阻断伤口内细胞转化生长因子 β(TGF β)的信号转导来抑制瘢痕形成的可行性。方法　用含TGF β截短型受体 (tTGF βRII)基因的重组腺病毒 (10 0空斑形成单位 /细胞 ,pfu/细胞 )感染正常人皮肤成纤维细胞 ,Northern印迹观察tTGF βRII表达对TGF β1基因表达的调控。将 β 半乳糖苷酶 (β gal)重组腺病毒 (1× 10 9pfu)和tTGF βRII重组腺病毒 (1× 10 9pfu)分别注入新生 10天的Sprague Dawley (SD)大鼠背部皮肤的左右侧 ,并在注射部位作线型切口 ,通过组织学检测和免疫组化观察tTGF βRII表达对抑制伤口瘢痕的作用和减少伤口炎症反应及TGF β1合成的作用。在tTGF βRII转基因小鼠和对照小鼠作同样线性切口来进一步证实tTGF βRII对抑制瘢痕的作用。结果　tTGF βRII在细胞内的过度表达显著抑制TGF β1的基因表达。腺病毒介导的皮肤基因转染在注射后 2～ 3天基因表达达高峰 ,5～ 7天逐渐下降。术后 2周 ,组织学检测发现tTGF βRII转基因小鼠伤口内相对瘢痕面积和对照小鼠伤口内相对瘢痕面积分别为 136 96 9 8± 6 6 339和 4 74 6 4 1 6±2 2 7396 ,转基因鼠瘢痕面积约为对照鼠的 2 9% (P <0 0 5 )。术后 2周 ,大鼠tTGFObjective TGF β plays a key role in wound scarring This study explored the possibility of using gene therapy to inhibit wound scarring by blocking TGF β signaling Methods In vitro , human normal dermal fibroblasts were infected with recombinant adenoviruses of truncated TGF β receptor II (tTGF βRII, 100 pfu/cell) and β galactosidase (β gal, 100 pfu/cell), and their effects on regulating TGF β1 gene expression were analyzed by Northern blot For gene therapy, β gal and tTGF βRII viruses (1×10 9 pfu)were injected intradermally at left and right side of dorsal skin of newborn Sprague Dawley rats ( n =15) respectively A full thickness incisional wound (0 5 cm long) was created at the injection sites of each rat 2 days post injection An incisional wound was similarly created in the middle part of the dorsal skin of tTGF βRII transgenic mice ( n =5) and control mice ( n =5) Wound tissues of rats and mice were harvested at various time points post wounding for histological and immunohistochemical analysis Scar area in tissue section was measured by Image Pro Plus software Results Over expression of tTGF βRII markedly reduced TGF β1 gene expression in dermal fibroblasts Adenovirus mediated gene expression in skin reached a peak level 2～3 days post injection, and decreased gradually at 5～7 days Two weeks post wounding, histology and quantitative analysis demonstrated that relative scar area in the wounds of transgenic mice and control mice were 136 969 8±66 339 and 474 641 6±227 396 respectively, the scar area of transgenic wounds was 29 percent of control area ( P <0 05) In all rats, wounds transfected with tTGF βRII gene healed with much less scarring (relative scar area 128 311 2±36 764 6) than control wounds (251 189 1±62 544 7) of the same rat, with a 45% reduction of scar area in average ( P <0 001) In addition, the tTGF βRII expression also decreased inflammation and TGF β1 production in treated wounds, and promoted the

关 键 词：转化生长因子Β 瘢痕 伤口愈合 实验研究 

分 类 号：R622[医药卫生—整形外科]