摘要目的 探讨鞘内注射趋化因子配体2(CCL2)中和抗体对胫骨癌痛大鼠机械痛行为及脊髓小胶质细胞活化的影响.方法 采用Walker 256乳腺癌细胞,构建大鼠胫骨癌痛模型.雌性SD大鼠(160～180 g)40只,随机分为5组(n=8):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+ control IgG组(Ⅳ组)和骨癌痛+CCL2中和抗体组(Ⅴ组).术后第10～12天,每天1次鞘内注射CCL2中和抗体或control IgG( 10 μg/15μ1).分别于术前、术后1d,3d,5d,7d,10 d,14 d,21 d测定各组大鼠机械痛敏阈值(PMWT).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各组大鼠脊髓背角小胶质细胞标志物(Iba-1)的平均光密度值(MOD).结果 (1)痛行为学:术后10d,14 d,21 d,Ⅲ组大鼠PMWT分别为:(1.78 ±0.38)g,( 1.70±0.17)g,(1.35±0.07)g；Ⅳ组大鼠PMWT分别为:(2.99±0.67)g,(2.52±0.75)g,(1.13±0.07)g;Ⅴ组大鼠PMWT分别为(5.88±0.66)g,(7.81±0.75)g,(6.19±0.53)g.与Ⅲ组相比,Ⅴ组大鼠PMWT值明显升高(P＜0.01),Ⅳ组大鼠PMWT值无明显变化(P＞0.05).(2)免疫荧光染色:术后14d,Ⅲ、Ⅳ、Ⅴ组大鼠脊髓背角Iba-1的MOD值分别为(151.3±10.8),(149.2±10.6),(74.5±5.0)；与Ⅲ组相比,Ⅴ组大鼠MOD值明显升高(P＜0.01)；Ⅳ组大鼠MOD值无明显变化(P＞0.05).结论 鞘内注射CCL2中和抗体能显著减轻胫骨癌痛大鼠的机械痛敏,并能抑制其脊髓小胶质细胞的活化.

abstractsObjective To investigate the role of CCL2 in pain facilitation and spinal mechanisms in the rat model of bone cancer pain.Methods The bone cancer pain model was developed by inoculating.Walker 256 mammary gland carcinoma cells into the rat tibia medullary cavity.SD female rats were divided into 5 groups randomly ( n =8):sham group( group Ⅰ),sham + CCL2 antibody group( group Ⅱ),BCP group( group Ⅲ),BCP +control lgG group ( group Ⅳ),BCP + CCL2 antibody group ( group Ⅴ ).VonFrey threshold was measured one day before operation and 1 st,3 rd,5th,7th,10th,14th,21 st after operation.CCL2 antibody or control lgG was injected intrathecally from 10th to 12th day.The expression of the spinal Iba-1 ( microglial marker) in rat lumbar4-5 was detected by immunohistochemistry assay.Results From the 10th to 21st day after operation,the PMWT of group Ⅲ rats were ( 1.78 ±0.38)g,( 1.70 ±0.17)g,( 1.35 ±0.07 )g;group Ⅳ rats were (2.99 ±0.67)g,(2.52 ±0.75)g,(1.13±0.07)g ; and group Ⅴ rats were (5.88±0.66)g,(7.81 ±0.75)g,(6.19±0.53)g.Compared with group Ⅲ,the PMWT of group Ⅴ was remarkly higher (P＜0.01) ; group Ⅳ had no obvious statistical significance (P＞0.05).At the 14th day after operation,the MOD of group Ⅲ,Ⅳ and Ⅴ rats were (151.3 ±10.8 ),( 149.2 ± 10.6),(74.5 ± 5.0),Compared with group Ⅲ,the MOD of group Ⅴ was significantly increased (P＜0.01 ),group Ⅳ had no obvious statistical significance (P ＞ 0.05 ).Conclusion Intrathecal injection of CCL2 antibody can remarkly attenuate established pain facilitation of tibial bone cancer pain rats,and significantly suppress the expression of Iba-1.It suggests that CCL2 is involved in the bone cancer pain via activation of spinal microglia.