摘要
目的 了解Ⅰ型前胶原基因反义寡聚核苷酸对人增生性瘢痕的作用 ,探讨增生性瘢痕的基因治疗。 方法 选择 12 0只裸鼠建立增生性瘢痕动物模型 ,将模型随机分为空白对照组(C组 ,6只 )、RPMI16 40组 (R组 ,6只 )、反义寡聚核苷酸 1(ASONs1)治疗组 (5 4只 )及反义寡聚核苷酸 2 (ASONs1)治疗组 (5 4只 ) ,后两组又分为 2 5 ,5 0 ,10 0 μg 10 0 μl三个剂量组 ,每个剂量组又分为治疗 7,10 ,14d 3组 ;分别合成位于Ⅰ型前胶原基因 5’端翻译区域 2 1bp的ASONs1和位于第 1个外显子与第 1个内含子之间 2 2bpASONs2 ,用微注射法分别将两基因片段作用于动物模型 ,通过测定增生性瘢痕Ⅰ型胶原含量变化、瘢痕体积变化以及运用光、电镜研究不同剂量反义寡聚核苷酸的抑制作用。 结果 ASONs1和ASONs2能使瘢痕体积缩小 ,在治疗 10 ,14d后 ,瘢痕组织中Ⅰ型胶原含量降低 ,与C组和R组比较 ,差异有显著性意义 (P <0 .0 5 ) ;光、电镜下见瘢痕结构疏松 ,胶原纤维变小 ,成纤维细胞的粗面内质网和线粒体减少 ,而C组和R组无明显变化。 结论 ASONs1和ASONs2能够有效抑制Ⅰ型胶原蛋白的合成 。
Objective To investigate the effect of antisense oligonucleotide of type Ⅰ procollagen on hypertrophic scar and to explore a new gene therapy to hypertrophic scar. Methods The scar models were established in 120 nude mice by a microinjection of ASONs1 and ASONs2. All the mice were divided into blank control group (6 mice), RPMI1640 group (6 mice), ASONs1 treatment group (54 mice) and ASONs2 treatment group (54 mice). The latter 2 groups were subdivided into 25, 50 and 100 μg/μl groups which were further assigned into 3 groups, i.e., 7, 10 and 14 days after treatment. The phosphorothioate ASONs1 and ASONs2 for type Ⅰ collagen used in this study were complementary to type Ⅰ collagen mRNA at the translationregion (21 bp) and at the region between exonI and introx (22 bp). We observed the effect of antisense oligonucleotide at 7,10 and 14 days after gene transfer. Results Both ASONs1 and ASONs2 could inhibit type Ⅰ collagen of hypertrophic scar. The scar volume was significantly reduced in treatment groups. Under the light microphone and electron microscope, the structure of scar became loose, collagenous fibre diminished, while the mitochondrion and rough endoplasmic reticulum in fibroblast reduced. There were no remarkable changes in control group and blank group. Conclusions The results suggest that antisense ASONs1 and ASONs2 can inhibit type Ⅰ collagen expression and hence suppress hypertrophic scar.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2001年第6期327-329,共3页
Chinese Journal of Trauma
基金
广东省医学联合攻关基金资助项目 ( 9890 0 4)
