促红细胞生成素治疗大鼠腰椎间盘突出症动物模型的实验研究

Treatment of Lumbar Intervertebral Disc Herniation with Erythropoietin in a Rat Model

目的建立大鼠腰椎间盘突出症的模型,探讨促红细胞生成素治疗大鼠腰椎间盘突出症动物模型的疗效。方法将30只SD大鼠按随机数字表法分为2组:实验组18只和对照组12只。2组大鼠分别建立腰椎间盘突出症模型。首先用自体髓核移植至L4、L5脊神经根,然后实验组大鼠将含促红细胞生成素的明胶海绵固定在神经根旁后缝合切口,对照组大鼠不作特别处理缝合切口。模型建立成功后,实验组大鼠采用促红细胞生成素5 000 U.kg-1.次-1.d-1腹腔注射,对照组大鼠采用生理盐水1 mL.次-.1d-1腹腔注射。术后12、24、36、48、72、96 h各处死实验组大鼠3只、对照组大鼠2只,取受压神经根处的神经及周围组织分别进行TUNEL染色观察细胞凋亡及TNF-α阳性细胞数。结果实验组大鼠术后24、48、72、96 h脊神经细胞凋亡率、术后12、24、36、48 h TNF-α阳性细胞数均显著低于对照组,差异均有统计学意义(均P<0.05)。结论促红细胞生成素治疗大鼠腰椎间盘突出症动物模型能抑制神经细胞的凋亡及TNF-α的表达,并能取得较好的疗效。

Objective To establish a rat animals model of lumbar intervertebral disc herniation, and to explore the curative effect of the erythropoietin on lumbar intervertebral disc herniation. Methods Thirty SD rats were randomly divided into experiment group （n=18）and control group （n=12）. For the disc herniation model,autologous nucleus pulposus was applied to L4,L5 nerve roots.In the experiment group,incisions were sutured after gelatin sponge containing erythropoietin was fixed next to the nerve roots.The control rats only received incision suturing. After successful establishment of the model,rats in experiment group and control group were intraperitoneally injected with erythropoietin（5 000 U. kg-1） and normal saline （1 mL）once per day,respectively.At 12,24,36,48,72 and 96 hours after operation, 3 rats in experiment group and 2 in control group were sacrificed to take nerves and surrounding tissues for TUNEL staining and TNF-α positive cell counting. Results The apoptosis rate of spinal nerve cells at 24,48,72 and 96 hours after operation and the number of TNF-α positive cells at 12,24,36 and 48 hours after operation in experiment group were significantly lower than those in control group （all P〈0.05）. Conclusion The erythropoietin can effectively inhibit nerve cell apoptosis and TNF-α expression in lumbar intervertebral disc herniation animals model.