艾芬地尔重复鞘内注射对骨癌痛模型小鼠痛行为的影响

Effect of repeated intrathecal injection of ifenprodil on pain behaviors in mice with bone cancer pain

目的观察艾芬地尔重复鞘内注射对小鼠骨癌痛的影响。方法96只雄性C3H／HeJ小鼠随机分为肿瘤给药组（T组）、肿瘤对照组（c组）和假手术组（S组）。将含2×10^5个纤维肉瘤NCTC2472细胞的最小必需培养基（Ⅱ．MEM）注射到小鼠右侧股骨远端骨髓腔内，制作骨癌痛模型。假手术组注入不含肿瘤细胞的α-MEM。T组在术后第14天开始，鞘内注射艾芬地尔5μl（10μg），每天1次，连续3天。C组和s组在相同时间点分别给予相同体积的溶媒。各组于接种前1天，接种后第3天、第5天、第7天、第10天、第14天、第17天、第19天和第23天检测痛行为学指标，包括小鼠的自发抬足次数、机械性缩足反射阈值（PWMT）和热缩足反射潜伏期（PwTL）。按照分组在相应时间点处死小鼠，用Western-blotting方法检测脊髓腰膨大NR2B蛋白表达水平。结果肿瘤细胞接种后第14天，和s组及术前基础值相比，T组小鼠自发性抬足[（12．88±1．64）次]显著增加，PWMT（0．39±0．17）g和PWTL（11．59±1．67）S明显下降（P〈0．05）。脊髓NR2B蛋白（2．12±0．13）显著增加（P〈0．05）。肿瘤细胞接种后第17天，第19天，第23天，和第14天及C组相比，T组自发性抬足[（5．13±1．38）次，（4．70±1．58）次，（5．64±1．17）次]明显减少，PWMT[（1．10±0．65）g，（0．95±0．56）g，（1．05±0．26）g]和PWTL[（15．17±1．27）s，（15．93±2．18）s，（16．28±1．48）S]显著增加（P〈0．05）。脊髓NR2B蛋白[（1．42±0．17），（1．67±0．53），（1．14±0．79）]显著减少（P〈0．05）。结论重复鞘内注射艾芬地尔能显著改善骨癌痛小鼠的痛行为，降低脊髓NR2B蛋白。

Objective To investigate the effect of ifenprodil in the mice of bone cancer pain. Methods 96 male C3 H/HeJ mice were divided randomly into tumor group （Group T）, control group （Group C ） and sham group（ Group S）. The a-minimal essence media （α-MEM） with 2 × 10^5 osteosarcoma NCTC 2472 cells were implanted into the intramedullary space of the right femurs of mice to induce ongoing bone cancer related pain behaviors. The sham group was inoculated by oL-MEM without any cells. On the 14th d after inoculation, pain ethol- ogy indexes such as the spontaneous lifting behaviors, the paw withdrawal mechanical threshold（PWMT） and the paw withdrawal thermal latency（ PWTL） were observed on 1 d before inoculation and on 3 d,5 d,7 d, 10 d, 14 d, 17 d, 19 d ,23 d after inoculation. Lumbar intumescentia of mice in each group were taken out to investigate the expression level of NR2B western blot after pain behaviors tests at the same time point after intrathecal injection. Results （1） At dayl4 after the operation, the obvious increasing of spontaneous lifting behaviors （（ 12.88 ±1.64 ） ） and the expression of N R2B （2.12 ± 0.13 ）, the significant decreasing of PWMT （ （0.39 ±0.17 ）g） and PWTL（ （ll. 59 ± 1.67）s） were observed in group T compared with group S and preoperative base level（P〈 0.05 ）. （2） At day 17 ,day 19 and day 23 after the operation,compared with the basal level of day14 before admin- istration and group C,the spontaneous lifting behaviors （ （5.13±1.38） , （4.70 ± 1.58） , （5.64 ± 1. 17） ） of group T were obviously decreased, PWMT （ （ 1. 10 ± 0.65 ） g, （0.95 ±0.56 ） g, （ 1.05 ± 0.26 ） g） and PWTL （（15.17±1.27）s,（15.93 ±2. 18）s,（16.28 ±1.48）s） were increased, the expression of NR2B（（1.42 ± 0.17） ,（1.67 ±0.53） ,（1.14 ±0.79）） were significantly decreased（P〈0.05）. Conclusion Repeated intrathca1 injection of ifenprodil can efficiently relieve spontaneous lifting behaviors, mechanical hyperalgia and thermal hyperalgia and decrease the expression of lumbar intumescentia NR2B in the mouse model of bone cancer pain.