局部应用羟基脲预防大鼠硬膜外纤维化的实验研究

Experimental study in prevention of epidural fibrosis in rats by local application of hydroxyurea

  • 摘要:
    目的 探讨大鼠椎板切除术后预防性局部使用羟基脲(HU)的疗效。
    方法 选取8周龄SD大鼠36只,随机分为A组(对照)、B组(15 mg/mL HU)、C组(30 mg/mL HU)。制备椎板切除术模型; 暴露硬脊膜后, A组生理盐水冲洗后逐层关闭术区,B、C组分别以浸湿不同浓度的HU棉垫放入术区内5 min后移除,逐层闭合术区。观察手术前后大鼠脊髓评分的变化。术后4周,通过一般观察、苏木精-伊红(HE)染色、成纤维细胞计数、天狼猩红染色和免疫组织化学染色来评估术后硬膜外粘连情况。
    结果 各组手术前后的大鼠脊髓评分比较,差异无统计学意义(P>0.05)。大体观察和HE染色显示,与A组相比, B、C组大鼠术区硬膜外粘连程度明显减轻。采用Image J对各组成纤维细胞计数,与A组相比,B、C组成纤维细胞数量呈浓度依赖性下降,差异有统计学意义(P < 0.001或P < 0.000 1); 与B组比较, C组成纤维细胞数量更低,差异有统计学意义(P < 0.01)。天狼猩红染色显示B、C组中Ⅰ型胶原纤维显著减少, Ⅲ型胶原纤维表达增多。
    结论 大鼠椎板切除术后局部应用HU可以减缓硬膜外纤维化的发生。

     

    Abstract:
    Objective To explore the efficacy of postoperative topical preventive application of hydroxyurea (HU) in the rats with laminectomy.
    Methods A total of 36 8-week SD rats were selected and randomly divided into group A (controls), group B (15 mg/mL HU) and group C (30 mg/mL HU). A laminectomy model was prepared; after exposing the dura mater, the surgical area was closed layer by layer after flushing with physiological saline in the group A, while in the group B and the group C, different concentrations of HU cotton pads were soaked and placed in the surgical area for 5 minutes before removal, and the surgical area was closed layer by layer. Change in spinal cord scores of rats before and after surgery was observed. At 4 weeks after surgery, the postoperative epidural adhesions were evaluated through macroscopic observation, hematoxylin eosin (HE) staining, fibroblast counting, Sirius red staining and immunohistochemical staining.
    Results There was no significant difference in the spinal cord scores of rats before and after surgery among the three groups (P>0.05). Macroscopic observation and HE staining showed that compared with the group A, the degrees of epidural adhesion in the surgical area of rats in the group B and the group C were significantly reduced. Image J was used to count fiber cells in each group, and compared with the group A, the number of fibroblasts in the group B and the group C decreased significantly in a concentration dependent manner (P < 0.001 or P < 0.000 1); compared with the group B, the number of fibroblasts in the group C was significantly lower (P < 0.01). Sirius red staining showed a significant decrease in type Ⅰ collagen fibers expression and a significant increase in type Ⅲ collagen fiber expression in the group B and the group C.
    Conclusion Postoperative topical application of HU in the rats with laminectomy can slow down the occurrence of epidural fibrosis.

     

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