骨癌痛大鼠脊髓NF-κB、IL-6和TNF-α表达的变化
Changes in expression of NF-κB, IL-6 and TNF-α in spinal cord in a rat model of bone cancer pain
摘要目的 评价骨癌痛大鼠脊髓NF-κB、IL-6和TNF-α表达的变化.方法 雌性健康SD大鼠72只,体重150~180 g,随机分为3组(n=24):对照组(C组)、假手术组(S组)和骨癌痛组(BP组).BP组于大鼠左侧胫骨干骺端骨髓腔内注射5μl Walker 256(1×105)乳腺癌细胞制备骨癌痛模型,S组左胫骨上端注射Hank液5μl.分别于肿瘤细胞接种前、接种后2、4、6、9、12、14、16、18和21 d时测定机械阈值.分别于肿瘤细胞接种后6、12和18 d时,处死3只大鼠,取L4-6脊髓组织,采用RT-PCR法测定NF-κB p65 mRNA、IL-6 mRNA和TNF-α mRNA的表达;另处死3只大鼠,取L4-6脊髓组织,计数NF-κBp65阳性细胞,采用免疫组化法测定NF-κB p65表达.结果 与C组和S组比较,BP组机械痛阈降低,脊髓NF-κB p65及其mRNA、IL-6 mRNA和TNF-αmRNA的表达上调,NF-κB p65阳性细胞计数增加(P<0.05或0.01);C组和S组间上述指标比较差异无统计学意义(P>0.05).结论 大鼠胫骨骨髓腔内注射肿瘤细胞通过激活脊髓NF-κB,导致炎性细胞因子IL-6和TNF-α大量释放,从而诱发骨癌痛.
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abstractsObjective To evaluate the changes in expression of NF-κB, IL-6 and TNF-α in spinal cord in a rat model of bone cancer. Methods Seventy-two female SD rats weighing 150-180 g were randomly divided into 3 groups (n = 24 each): control group (group C);sham operation group (group S) and bone cancer pain group (group BP). Bone cancer was induced by intra-tibial inoculation of 1 × 105 Walker 256 breast cancer cells. Paw withdrawal threshold to mechanical stimulation was measured with yon Frey filaments. The expression of NF-κB p65, IL-6 and TNF-α mRNA in the spinal cord was determined by RT-PCR and the expression of NF-κB p65 by immuno-histochemistry and NF-κB p65 positive cell count was determined. Results The paw withdrawal threshold was significantly lower and the expression of NF-κB p65, NF-κB p65 mRNA, IL-6 mRNA, TNF-α mRNA and NF-κB p65 positive cell count in the spinal cord were significantly higher in group BP than in group C and S ( P <0.05 or 0.01 ). Conclusion Intra-tibial inoculation of Walker 256 breast cancer cells activates NF-κB in the spinal cord, leading to the increased release of IL-6 and TNF-α and mechanical hyperalgesia.
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