透明质酸钠对硬膜外瘢痕中TGF-β3和Smad6基因表达的影响  

Effects of sodium hyaluronate on the expression of TGF-β3 and Smad 6 of the epidural scar tissue

作　　者：郑悦亮[1] 夏志敏[2] 周辉[2] 赵万军[2] 

机构地区：[1]浙江省人民医院急诊科,杭州310014 [2]杭州市中医院骨伤科

出　　处：《中华急诊医学杂志》2008年第9期956-960,共5页Chinese Journal of Emergency Medicine

基　　金：浙江省医药卫生科研基金项目（2007A083）;杭州市医药卫生科技计划项目（2007A027）

摘　　要：目的探讨透明质酸钠对硬膜外瘢痕中转化生长因子（transforming growth Factor，TGF）-β3和Smad 6表达水平的影响。方法雄性SD大鼠60只随机分成空白对照组（A）、透明质酸钠组（B），每组30只。手术切除第4、5腰椎椎板，造成0．25cm×1．00cm硬脊膜裸露区，B组于硬膜囊外均匀地涂上一层透明质酸钠约0．3ml，A组以等量生理盐水处理。术后第2、4、8周末处死动物，逆转录酶链反应（Reverse Transcription Polymerase Chain Reaction，RT-PCR）测定其硬膜外瘢痕组织中TGF-β3和Smad 6的表达水平，按Rydell法评定粘连程度，透射电镜观察其超微结构。大体观察数据的比较用两个独立样本比较的秩和检验（Mann—Whitney U检验）；TGF-β3和Smad 6基因表达量的数据（x^±s）用完全随机设计两样本均数的t检验。结果第2周时，两组TGF-β3 mRNA表达均较低，均数分别为0．22和0．257（P=0．027）；第4周时，B组TGF-β3的表达明显增高，两组均数分别为0．362和0．411（P=0．006）；第8周时，A组TGF-β3 mRNA的表达水平虽也有明显增高，但两组间仍存在明显的差异，分别为0．427和0．470（P=0．015）。Smad 6 mRNA的表达趋势与TGF-β3类似，第2周、第4周、第8周分别为0．169和0．205（P=0．089）、0．294和0．351（P=0．031）、0．469和0．543（P=0．021）。大体观察也发现B组粘连程度明显降低（P〈0．05）。透射电镜示各时相B组瘢痕中成纤维细胞以及细胞内的细胞器的数量均减少，胶原纤维的合成受到抑制。结论透明质酸钠通过上调TGF-β3和Smad 6的表达，减少了成纤维细胞增殖和胶原等细胞外基质（Extracellular Matrix，ECM）的沉积，达到了预防硬膜外瘢痕增生、粘连的效果。Objective To study the effect of sodium hyaluronate on the expression of transforming growth factor-β3 （TGF-β3） and Smad 6 of the epidural scar tissue. Method Totally 60 male SD rats were randomly divided into two groups：the control group（A, n = 30）, sodium hyaluronate group（B, n = 30）. 0.25 × 1 cm^2 dura mater uncovered area laminectomy was performed at L4 and L5, covered with 0.3 ml sodium hyaluronate in group B, covered with same amount of saline in group A. The animals were sacrificed at 2, 4 and 8 weeks after operation. The specimens were prepared for determination of the expression of TGF-β3 and Smad 6 at scar tissue and the degree of scar adhesion according to Rydell method, and observed the ultrastructure changes of scar tissue with transmission electron microscope. Results At 2 weeks after operation, the expression of TGF-β3 mRNA of two groups were 0.22 and 0.257 （ P = 0. 027）, respectively. At 4 weeks, group B was was increased significantly, and the mean numbers were 0. 362 and 0. 411 （ P = 0. 006）. At 8 weeks, the expression of TGF-β3 mRNA of group A was increased significantly, too, but the difference between two groups was significantly, they were 0. 427 and 0. 470 （ P = 0. 015）, respectively. The trend of the expression of TGF-β3 and Smad 6 mRNA was similar. At 2, 4, 8, weeks, the expression was 0.169 and O. 205 （ P = 0. 089）, 0. 294 and 0.351 （ P = 0.031 ）, and 0. 469 and 0. 543 （ P = 0. 021 ）, respectively. In group B the duramater adhesion was decreased （ P 〈 0. 05）, the proliferation of fibroblasts and fibroblastic function were inhibited （ P 〈 0. 05）. Conclusions Sodium hyaluronate up-regulated the expression of TGF-β3 and Smad 6 and reduced the proliferation and collagen synthesis in fibroblast culture in the scar tissue.

关 键 词：椎板切除术 瘢痕 转化生长因子-Β3 SMAD 6 

分 类 号：R622[医药卫生—整形外科]