趋化因子CCL2中和抗体在大鼠胫骨癌痛治疗中的作用

Role of chemokine CCL2 antibody in the treatment of tibia cancer pain in rats

目的观察鞘内注射趋化因子CCL2中和抗体在胫骨癌痛大鼠治疗中的作用,并探讨可能的机制。方法 40只雌性SD大鼠随机均分为五组:假手术组(Ⅰ组)、假手术+CCL2中和抗体组(Ⅱ组)、骨癌痛组(Ⅲ组)、骨癌痛+controlIgG组(Ⅳ组)和骨癌痛+CCL2中和抗体组(Ⅴ组)。Walker256乳腺癌细胞注入胫骨骨髓腔建立大鼠胫骨癌痛模型。术后10～12d鞘内注射CCL2中和抗体或对照IgG(10μg/15μl)。测定术前1d,术后1、3、5、7、10、14、21d各组大鼠自由行走痛评分。另取30只大鼠,分组同前(n=6),术后14d取材,免疫荧光染色法测定脊髓背角星形胶质细胞标志物(GFAP)的平均光密度值(MOD)。结果与Ⅲ组相比,Ⅴ组大鼠术后10、14和21d自由行走痛评分明显降低,术后14dMOD值明显降低(P<0.01),Ⅳ组对应时点各值无明显变化。结论胫骨癌痛大鼠鞘内注射CCL2中和抗体能显著减轻自由行走痛并抑制脊髓星形胶质细胞的活化。CCL2可能通过激活脊髓星形胶质细胞参与大鼠胫骨癌痛维持的调控。

Objective To observe the role of Chemokine CCL2 antibody in the treatment of tibia cancer pain in rats and explore its possible mechanism. Methods Forty SD female rats were equally randomized into 5 groups： sham operation group （group I ）, sham operation＋CCL2 antibody group （group Ⅱ）, bone cancer pain （BCP） group （groupⅢ）, BCP＋control IgG group （group IV）, BCP＋ CCL2 antibody group （group V ）. The tibia cancer pain model was established by inoculating Walker 256 mammary gland carcinoma cells into the rat tibia medullary cavity. CCL2 antibody or control IgG was injected intrathecally after 10 th and 12th day. The score of walk-associated ambulatory pain was measured one day before operation and 1, 3, 5, 7, 10, 14, 21 day after operation. Another 30 rats were equally randomized into the same groups and sacrificed 14 days after operation. The mean optical density （MOD） of the spinal GFAP （astrocyte marker） in rat spinal cord was detected by immunohistochemistry assay. Results Compared with groupⅢ, the score of walk-associated ambulatory pain of group V was remarkly lower on 10, 14 and 21 d after operation and the MOD was significantly decreased on 14 d after operation （P 〈 0. 01 ）; group IV had no obvious statistical significant difference on the score and the MOD. Conclusion Intrathecal injection of CCL2 antibody attenuates the score of walk-associated pain and suppresses the activation of GFAP in tibial cancer pain rats. It suggestes that CCL2 is involved in the bone cancer pain via activating spinal astrocyte.