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目的观察骨癌痛模型大鼠患侧腰段脊髓 p38α和 p38p 的分布及细胞定位,探讨其在骨癌痛中的作用机制。方法选择雌性 SD 大鼠20只,随机分为2组,每组10只:A 组(对照组):左侧胫骨上段骨髓腔注入3μl Hanks 液;B 组(模型组):左侧胫骨上段骨髓腔注入3μl MADB-106大鼠乳腺癌细胞(4.8×10~3/μl)。术前及术后14 d,各组大鼠隔日观察机械痛及辐射热痛阈值变化。第14 d,取大鼠脊髓14～6节段,采用免疫组织化学 ABC 法和荧光双标法观察骨癌痛模型大鼠患侧腰段脊髓 p38α和 p38β免疫反应阳性产物的分布变化和细胞定位。结果 A 组大鼠对机械、辐射热痛刺激的缩爪阈值在术后各时间点组内相比,差异无统计学意义;B 组大鼠对辐射热痛刺激缩爪阈值在术后的前6 d 与 A 组相比,差异有统计学意义(P<0.05);术后第14 d,与 A 组大鼠对机械痛刺激缩爪阈值和辐射热痛阈值相比,B 组差异有统计学意义(P<0.05)。B 组大鼠患侧腰段脊髓背角Ⅰ～Ⅳp38α和 p38β免疫反应吸光值明显高于 A 组,两组相比差异有统计学意义(均 P<0.05)。荧光双标显示患侧腰段脊髓背角 p38α与神经元特异性核蛋白标志物 NeuN 有共表达,p38β与小胶质细胞标志物 OX-42有共表达。结论脊髓 p38α和 p38β参与了骨癌痛的发生和发展,p38α主要在脊髓神经元表达,而 p38β则在脊髓小胶质细胞中表达。
Objective To observe the distribution and cellular localization of p38α and p38p in the ipsilateral lumbar spinal cord of rat model of bone cancer pain and to explore its mechanism in bone cancer pain. Methods Twenty female Sprague-Dawley rats were randomly divided into 2 groups (10 rats in each group): A group (control group): 3 μl Hanks solution was injected into the left medial tibial bone marrow; B group (model group): the left upper tibia bone marrow 3 μl of MADB-106 rat breast cancer cells (4.8 × 10 -3 / μl) were injected into the lumen. Preoperative and postoperative 14 d, the rats in each group were observed mechanical pain and radiation pain threshold changes every other day. On the 14th day, the 14 ~ 6 segments of the spinal cord of rats were taken and the distribution and cell localization of p38α and p38β immunoreactive products in ipsilateral lumbar spinal cord of the bone cancer pain model were observed by immunohistochemical ABC and fluorescent double labeling . Results There was no significant difference in the paw withdrawal thresholds of rats in group A between mechanical and radiation thermal pain stimuli at different time points after operation. In group B, Compared with group A, the difference was statistically significant (P <0.05) on the 6th day and the group A on the 14th day Significance (P <0.05). The immunoreactivity of Ⅰ ~ Ⅳp38α and p38β in the ipsilateral lumbar spinal dorsal horn of group B was significantly higher than that of group A (P <0.05). Fluorescence double labeling showed that the ipsilateral spinal cord dorsal horn p38α co-expressed with neuron-specific nuclear protein marker NeuN, and p38β co-expressed with microglial marker OX-42. Conclusion The spinal cord p38α and p38β are involved in the occurrence and development of bone cancer pain. P38α is mainly expressed in spinal cord neurons, while p38β is expressed in spinal cord microglia.