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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 ,从而抑制瘢痕增生
Objective To investigate the effect of type Ⅰ procollagen antisense oligonucleotide on human hypertrophic scars and to explore the gene therapy of hypertrophic scars. Methods A model of hypertrophic scar was established in 120 nude mice. The model was randomly divided into blank control group (C group, 6 mice), RPMI1640 group (R group, 6 mice), antisense oligonucleotide 1 ASONs1 treatment group (54) and antisense oligonucleotide 2 (ASONs1) treatment group (54). The latter two groups were divided into three groups of 25, 50 and 100 μg Group, and each dose group was divided into treatment groups of 7, 10, 14d 3; ASONs1 located at 2 1bp in the translation region of type Ⅰ procollagen gene 5 and endonuclease 1 Two 2bpASONs2 were injected into the animal model by microinjection. The content of type Ⅰ collagen in hypertrophic scar and the change of scar volume were determined. The inhibition of different doses of antisense oligonucleotide effect. Results ASONs1 and ASONs2 could reduce the size of scar. At the 10th and 14th day after treatment, the content of type Ⅰ collagen in scar tissue decreased. Compared with C and R group, the difference was significant (P <0.05) Electron microscopy showed loose scars, collagen fibers smaller, fibroblasts rough endoplasmic reticulum and mitochondria decreased, while C and R group no significant change. Conclusion ASONs1 and ASONs2 can effectively inhibit the synthesis of type Ⅰ collagen, thereby inhibiting the proliferation of scar