摘要
目的:研究鞘内注射(it) NF-κ B活化抑制剂二硫代氨基甲酸吡咯烷(pyrrolidme dithiocarbamate,PDTC)对骨癌痛大鼠机械痛敏及脊髓NF-κB表达的影响,探讨NF-κ B在大鼠骨癌痛中的作用.方法:按照本实验室建立的大鼠胫骨癌痛模型建模.雌性SD大鼠96只,体重150~180 g,按照数字表随机化的原则将大鼠分为4组(n=24):正常对照组(N组)、骨癌痛组(BP组)、生理盐水组(S组)和NF-κB抑制剂组(PDTC组).BP组于大鼠左侧胫骨干骺端骨髓腔内注射5μlWalker256(1×10^5)乳腺癌细胞制备骨癌痛模型,N组仅左胫骨上端注入5 μl Hank's液.于接种后9d鞘内注射生理盐水10 μl或PDTC 20 μg/10μl(生理盐水溶解),2次/d,共3d.于接种前、后2、4、6、9和12d、鞘内给药后1、3、6、12、24 h采用von Frey丝测定大鼠术侧后爪机械缩足反射阈值(paw withdrawal threshold,PWT),PWT测定结束后处死大鼠,取L4~6脊髓,采用real time-PCR法测定NF-κ B p65 mRNA表达,免疫组化法测定NF-κ B p65蛋白表达.结果:接种后6dBP组大鼠的PWT值与基础值相比开始降低,且差异有统计学意义(P<0.05),随着接种时间的延长其下降更加显著.鞘内注射PDTC可显著提高骨癌痛大鼠的PWT值,且与没注射PDTC前PWT值比,差异有统计学意义(P<0.05).与接种后6d时相比较,骨癌痛大鼠脊髓NF-κ B p65 mRNA和蛋白质的表达在接种后9d (P<0.01)和12d(P<0.01)时均显著上调,而鞘内注射PDTC可显著降低大鼠脊髓NF-κ B p65 mRNA和蛋白质的表达(P<0.01).结论:鞘内注射PDTC可减轻大鼠骨癌痛引起的痛敏,NF-κ B的激活可诱导或促进大鼠的胫骨癌痛.
Objective : To investigate the effect of intrathecal (i.t.) pyrrolidine dithiocar-bamate (PDTC, a NF- κ B inhibitor) on the mechanical hyperalgesia and the expression of NF- κ B in the dorsal horn of spinal cord in rats with bone cancer pain, and evaluate the role of NF- κ B in the mechanism of bone cancer pain. Methods: The rat tibial cancer pain model was established in our laboratory. 96 female SD rats weighing 150- 180 g were randomly divided into 4 groups (n = 24): normal control group (group N), bone cancer pain group (group BP), saline group (group S) and NF- κ B inhibitor group (group PDTC). Bone cancer was induced by intra-tibial inoculation of 1 × 10^5 Walker 256 breast cancer cells in group BP. Group N only received 5 μl Hank's liquid via injection through the upper left tibia. At day 9 after the inoculation, 10 μl normal saline or 20 μg/10 μl PDTC (dissolved in normal saline) was administered twice a day for three days. Paw withdrawal threshold (PWT) to mechanical stimulation was measured with yon Frey filaments beforeinoculation and 2, 4, 6, 9,12 d after inoculation as well as 1, 3, 6, 12, 24 h after intrathecal administration. Rats were sacrificed after PWT measurement and L4- 6 spinal cords were collected to determine the expression of NF- κ B p65 mRNA by real time-PCR and the expression of NF- κ B p65 protein by immunohistochemistry. Results: The PWT significantly decreased in group BP at day 6 after inoculation compared to baseline (P 〈 0.05). It further decreased with the prolonging of inoculation. The PWT value significantly increased after intrathecal injection of PDTC compared to the value before the injection (P 〈 0.05). Compared to 6 d after inoculation, the expression levels ofNF- κ B p65 mRNA and protein significantly increased at 9 d (P 〈 0.01) and 12 d (P 〈 0.01) after inoculation, indicating that intrathecal injection of PDTC could obviously inhibit the activation of NF- κ B p65 in the spinal cord induced by bone cancer pain (P 〈 0.01). Conclusion: Intrathecal injection of PDTC can alleviate the mechanical hyperalgesia induced by bone cancer pain. Activation of NF- κ B may induce or contribute to bone cancer pain.
出处
《中国疼痛医学杂志》
CAS
CSCD
北大核心
2014年第5期299-303,307,共6页
Chinese Journal of Pain Medicine
基金
国家自然科学基金(81171057)
江苏省自然科学基金(SBK201121879)
国家青年科学基金(H0903-81000479)
江苏省临床医学科技专项(重点病种的规范化诊疗研究)(SBL201220104)
关键词
NF-κB
骨肿瘤
疼痛
Nuclear factor-kappa B
Bone neoplasms
Pain