热休克蛋白47重组质粒对病理性瘢痕的体内干预研究

The effect of recombinant HSP47siRNA on the pathological scar in nude mice： in vivo study

目的利用RNAi技术通过热休克蛋白47重组质粒（HSP47siRNA）和脂质体的混合液对裸鼠病理性瘢痕动物模型的体内干预，分析HSP47基因在病理性瘢痕生成中的意义。方法构建裸鼠病理性瘢痕动物模型，第16天腹腔麻醉后实验组裸鼠在病理性瘢痕内注射质粒、脂质体混和液0．25ml，对照组裸鼠腹腔注射PBS液0．25ml，原笼饲养，7d回收标本，分别作mRNA水平、胶原蛋白水平以及免疫组化检测。结果对照组与实验组总胶原含量分别为（91．71±1．24）％和（82．12±4．79）％；实时荧光PCR检测HSP47 mRNA表达对照组和实验组分别为1042862．01±04194．36和306123．68±105857．08；Ⅰ型胶原蛋白mRNA表达对照组和实验组分别为10228 614．70±2532 879．04和6011 841．97±2886 897．17；对照组和实验组体积分别为（255．60±21．34）mm^3和（132．99±24．06）mm^3，上述指标检测结果组间比较差异均有统计学意义（P〈0．05）；而这些变化在增生性瘢痕组并没有出现，增生性瘢痕组胶原的变化、体积的变化均无统计学意义。结论采用RNAi技术，经过HSP47siRNA表达载体特异性沉默病理性瘢痕中HSP47基因表达后，瘢痕疙瘩中胶原蛋白的合成和分泌均能得到明显抑制，提示HSP47基因促进瘢痕疙瘩生成，并为抑制瘢痕疙瘩提供了新的靶点。

Objective To study the significance of HSP47 gene in the development of pathological scar. Methods The nude mice were used to reconstruct animal model of pathological scar. 16 days later, the mixture of recombinant HSP47 siRNA and liposome was injected into the pathological scar in experimental group. In the control group, 0.25ml PBS was injected intraperitoneally. 7 days after injection, the specimens were collected for detection of mRNA of HSP47, the collagen and for immunohistochemical study. Results In the control and experimental group, the collagen content was （91.71± 1.24）% and （82.12 ± 4.79）%, respectively; the expression of HSP47 mRNA was 1 042 862.01± 604 194.36 and 306 123.68 ±105 857.08, respectively; the expression of collagen ⅠmRNA was 10 228 614.70 ± 2 532 879.04 and 6 011 841.97 ± 2 886 897.17, respectively;the sear volume was （255.60 ± 21.34） mm^3 and （132.99±24.06） mm^3 , respectively. All the above results showed significant difference between the two groups （ P 〈 0.05 ） . Conclusions The collagen production can be reduced through suppression of the expression of HSP47 gene. It indicates that HSP47 gene enhance the development of keloid and could be used as the taget of treatment.