活血止痛汤对硬膜外瘢痕Smad2、Smad3和Smad4 mRNA表达的影响

The Effect of HuoXueZhiTong Recipe on the Expression of Smad2, Smad3 and Smad4 of the Epidural Scar Tissue

目的:探讨透明质酸钠和中药活血止痛汤预防硬膜外瘢痕形成时对Smad2、Smad3和Smad4表达水平的影响。方法:雄性SD大鼠90只随机分成空白对照组(A)、透明质酸钠组(B)、活血止痛汤组(C),每组30只。手术切除第4、5腰椎椎板,造成0.25cm×1cm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,其余两组以等量生理盐水处理。术后2周内C组予活血止痛汤2.5ml/kg灌胃;A组和B组予等量生理盐水。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Smad2、Smad3和Smad4的表达水平,碱水解法测定其硬膜外瘢痕组织中羟脯氨酸(Hydroxy proline,Hyp)的含量,按Rydell法评定粘连程度,透射电镜观察其超微结构。结果:①第2、4、8周时,B组和C组Smad2、Smad3和Smad4的表达水平均明显低于A组(P<0.05),尤其在第8周时,C组Smad3的表达水平与B组亦有统计学意义(P<0.05)。②第2周时,B组Hyp含量明显低于A组(P<0.05),而C组与A组差异无统计学意义;第4周时,与A组相比,B组和C组Hyp含量均较低(P<0.05)。第8周时,C组Hyp含量不仅低于A组(P<0.05),而且其与B组亦有显著性差异(P<0.05)。③大体观察也发现B组和C组粘连程度明显降低(P<0.05)。结论:透明质酸钠和活血止痛汤均可通过下调Smad2、Smad3和Smad4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 HuoXueZhiTong Recipe on the expression of Smad2, Smad3 and Smad4 of the epidural scar tissue. Methods ： Ninety male SD rats were randomly divided into three groups, the control group（A, n=30）, sodium hyaluronate group（B, n=30）, HuoXueZhiTong Recipe group（C, n=30）. 0.25 × 1cm^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 and C. First 2 weeks after operation, rats were fed with HuoXueZhiTong Recipe（2.5ml/kg） by gavage in group C, and with same amount of saline in group A and B. The animals were sacrificed in 2, 4 and 8 weeks after operation. The specimens were prepared for the determination of the expression of Smad2, Smad3 and Smad4 at scar tissue by a reverse transcription polymerase chain reaction （RT-PCR） , observed the content of Hydroxyproline （Hyp）, and evaluated the degree of scar adhesion according to Rydell method. Results.. In 2 , 4 , 8 weeks after operation, compared with Group A ,the expression of Smad2, Smad3 and Smad4 of group B and C were decreased （P〈0. 05）. Especially, in 8 weeks, the expression of Smad3 of group C was significant decreased compared with group B（P〈0.05）. In 2 weeks after operation, compared with Group A,the content of Hyp of group B was decreased （P〈0.05）, but group C compared with group A and B had no difference. In 4 weeks, compared with group A, the Hyp of group C was significantly decreased not only compared with group A（P〈0.05） but also with group B（P〈0.05）. In Group B and group C the duramater adhesion was decreased （P〈 0. 05）. Conclusion： HuoXueZhiTong Recipe down- regulated the expression of Smad2, Smad3 and Smad4 and reduced the proliferation and collagen synthesis in fibroblast culture in the scar tissue .