消退素D2对椎间盘突出所致根性神经痛的作用

The effect of resoivin D2on radicular pain caused by lumbar disc herniation

目的 观察消退素D2(RvD2)对非压迫性腰椎间盘突出症所致根性神经痛模型大鼠的影响。 方法 选取雄性Sprague-Dawley(SD)大鼠36只,按随机数字表法分为假手术组,模型组和RvD2组,每组大鼠12只。模型组和RvD2组均采用自体髓核组织填充法制作非压迫性腰椎间盘突出症模型,假手术组仅暴露相应手术部位,不作其它处理。造模成功后连续3d对假手术组和模型组均鞘内给予磷酸盐缓冲液(PBS)10μl,RvD2组则鞘内给予RvD2溶液10ng/10μl。于造模前1d和造模成功后连续7d观察3组大鼠术侧的50%缩足阈值(PWT),并于造模成功后第7天获取大鼠术侧L4至L6节段脊髓背角,采用蛋白质印迹法测定磷酸化的蛋白激酶B(p-AKT),蛋白激酶B(t-AKT),磷酸化的糖原合成激酶3β(p-GSK-3β)和糖原合成激酶3β(GSK-3β)的蛋白表达量,并用酶联免疫吸附试验测定肿瘤坏死因子α(TNF-α)、白介素-6(IL-6)、转化生长因子-β1(TGF-β1)和白介素-10(IL-10)的蛋白表达含量。 结果 造模成功后第1~7天,模型组和RvD2组大鼠50%PWT与假手术组同时间点比较,差异均有统计学意义(P<0.05);造模成功后第3~7天,RvD2组的50%PWT分别为(6.31±2.11)g、(7.37±1.58)g、(7.96±1.73)g、(8.46±1.55)g、(8.55±1.44)g,与模型组同时间点比较,差异均有统计学意义(P<0.05)。造模成功后第7天,模型组和RvD2组的p-AKT和p-GSK-3β蛋白阳性表达水平与假手术组比较,差异有统计学意义(P<0.05);且RvD2组的p-AKT和p-GSK-3β蛋白阳性表达水平与模型组比较,差异亦有统计学意义(P<0.05)。造模成功后第7天,模型组和RvD2组大鼠脊髓背角促炎细胞因子TNF-α和IL-6以及抗炎因子TGF-β1和IL-10的蛋白表达水平与假手术组比较,差异均有统计学意义(P<0.05); 且RvD2组促炎细胞因子TNF-α和IL-6以及抗炎因子TGF-β1和IL-10的蛋白表达水平与模型组比较,差异均有统计学意义(P<0.05)。 结论 RvD2可减轻非压迫性椎间盘突出大鼠的根性神经痛,其机制可能与抑制GSK-3β活性,下调促炎因子以及上调抗炎因子的蛋白表达水平相关。

Objective To explore the effect of resolvin D2 (RvD2) on radicular pain induced by intervertebral disc herniation. Methods Thirty-six male Sprague-Dawley rats were randomly divided into a sham operation group, a model group and an RvD2 group, each of 12. Non-compressive lumbar disc herniation was induced in the rats in the model and RvD2 groups using the autologous nucleus pulposus filling method. Those in the sham group had the surgical site exposed without any other treatment. After the modeling, 10 μl of phosphate-buffered saline solution was administered intrathecally to the rats in the sham and model groups for 3 days, while the rats in the RvD2 group received 10 ng/10 μl of RvD2 intrathecally as well. Paw withdrawal thresholds (50%PWT) were observed 1 day before modeling and 7 days afterward for the rats of all three groups. On the 7th day after modeling, the L4 to L6 spinal dorsal horns on the surgery side were resected to detect the protein expression of phosphorylated protein kinase B (p-AKT), protein kinase B (t-AKT), phosphorylated glycogen synthase kinase 3β (p-GSK-3β) and glycogen synthase kinase 3β (GSK-3β) using western blotting. The protein levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) were determined using enzyme-linked immunosorbent assays. Results On the 1st and 7th day after modeling, significant differences were observed between the model and sham groups in terms of the 50%PWT. From the 3rd day the average 50%PWT in the RvD2 group was significantly higher than that of the model group at the same time points. On the 7th day after the modeling the average p-AKT and p-GSK-3β protein levels of the model and RvD2 groups were significantly different from that of the sham group, and the model group′s average was also of significantly different from that of the RvD2 group. The average protein levels of the pro-inflammatory cytokines TNF-α and IL-6, as well as of the anti-inflammatory factors TGF-β1 and IL-10 in the dorsal horns of the model group and the RvD2 group were also significantly different on the 7th day, and both were significantly different from the sham group′s average. Conclusion RvD2 can alleviate radicular pain in rats with non-compressive disc herniation. The mechanisms might involve inhibition of GSK-3β activity, down-regulation of pro-inflammatory factors and up-regulation of anti-inflammatory factors.