脊髓趋化因子配体2在大鼠胫骨癌痛中的作用  

Role of chemokine ligand 2 in spinal cord in a rat model of tibia bone cancer pain

作　　者：徐幼苗[1,2] 申文[3] 陈晏[1] 岳红利 柳娇[1] 岳冬梅[1] 袁燕[3] 梁栋[3] 

机构地区：[1]江苏省麻醉学重点实验室,徐州市221002 [2]秦皇岛市第一医院疼痛科 [3]徐州医学院附属医院疼痛科 [4]河北省秦皇岛山桥医院麻醉科

出　　处：《中华麻醉学杂志》2011年第9期1052-1055,共4页Chinese Journal of Anesthesiology

基　　金：徐州市科技计划资助项目（XM08C068）

摘　　要：目的探讨脊髓趋化因子配体2（CCL2）在大鼠骨癌痛中的作用。方法健康成年雌性sD大鼠84只，体重160—180g，采用随机数字表法，将其随机分为3组（n=28）：正常对照组（c组）、假手术组（s组）和骨癌痛组（P组）。P组胫骨骨髓腔内注射Walker-256乳腺癌细胞混悬液制备大鼠胫骨癌痛模型；S组胫骨骨髓腔内注射生理盐水；C组不作任何处理。于接种前1d、接种后1、3、7、10、14和21d时，测定机械性刺激阈值。于接种前1d、接种后7、14和21d时痛阈测定结束后，各组随机处死6只大鼠，取L4-6脊髓组织，采用ELISA法测定CCL2含量，反映其表达。接种后14d时痛阈测定结束后，P组随机取4只大鼠，采用免疫荧光双标染色法观察脊髓背角CCL2与离子钙接头蛋白分子．1（小胶质细胞特异性标记物）、胶质纤维酸性蛋白（星形胶质细胞特异性标记物）和神经元特异核蛋白（神经元特异性标记物）的共表达情况。结果与C组和S组相比，P组接种后7—21d时机械性刺激痛阈下降，接种后7、14和21d时脊髓CCL2表达上调（P〈0．05）。骨癌痛大鼠脊髓背角CCL2在小胶质细胞和星形胶质细胞中存在表达，而在神经元中无表达。结论脊髓小胶质细胞和星形胶质细胞中释放的CCL2参与了大鼠胫骨癌痛的发生发展。Objective To investigate the role of chemokine ligand 2 （CCL2） in the spinal cord expression in a rat model of tibia bone cancer pain.Methods Eighty-four female SD rats weighing 160-180 g were randomly divided into 3 groups （ n = 28） ： control group （group C）, sham operation group （group S） and tibia bone cancer pain group （group P）. Tibia bone cancer pain was induced by intra-tibial inoculation of Walker-256 breast cancer cells. Paw withdral threshold to mechanical stimulation （MWT） was measured with von Frey filaments at 1 d before and at 1, 3, 7, 10, 14 and 21 d after inoculation. Six rats in each group were sacrificed after the measurement of MWT at 1 d before inoculation and at 7, 14 and 21 d after inoculation. Lumbar 4-6 segments of the spinal cord were removed for determination of the expression of CCL2 by ELISA. The coexpression of CCL2 with Iba-1 （a specific marker of microgha）, GFAP（a specific marker of astrocyte） and NeuN （a specific marker of neuron） was determined by double immunofluorescence assay after the measurement of MWT at 14 d after inoculation in group P. Results Compared with groups C and S, MWT was significantly decreased from 7 d to 21 d after inoculation, the expressive of CCL2 in the spinal cord up-regulated at 7, 14 and 21 d after inoculation in group P （P 〈 0.05）. CCL2 was expressed in the microglia and astrocyte but not in neuron in the spinal cord dorsal horn in a rat model of tibia bone cancer pain. Conclusion Release of CCL2 from microglia and astrocytes in the spinal cord was involved in mechanical hyperalgesia in a rat model of tibia bone cancer pain.

关 键 词：趋化因子CCL2 脊髓 骨肿瘤 疼痛 

分 类 号：R738.1[医药卫生—肿瘤]