脊神经结扎诱导神经病理痛大鼠脊髓背角BDNF的含量和表达上调  

UPREGULATION OF THE BRAIN-DERIVED NEUROTROPHIC FACTOR IN THE SPINAL DORSAL HORN OF NEUROPATHIC RATS INDUCED BY SPINAL NERVE LIGATION

作　　者：廖斐斐[1] 耿山景[1] 刘小旦[1] 党文豪[1] 蔡捷[1] 刘风雨[1] 万有[1] 韩济生[1] 邢国刚[1] 

机构地区：[1]北京大学神经科学研究所,基础医学院神经生物学系,教育部和卫生部神经科学重点实验室

出　　处：《中国疼痛医学杂志》2009年第3期162-165,170,共5页Chinese Journal of Pain Medicine

基　　金：国家自然科学基金(30370470);科技部国家重点基础研究发展计划"973"资助项目(2007CB512501);教育部留学回国人员科研启动基金(2008890)资助

摘　　要：目的：研究脊神经结扎（SNL）大鼠脊髓背角BDNF的含量和蛋白表达的变化,并探讨其在神经病理痛形成中的作用。方法：按照双盲随机的原则,将30只雄性SD大鼠随机分为SNL手术组和假手术对照组,每组各15只大鼠,进行酶联免疫吸附实验（ELISA）和免疫组化染色,检测神经病理痛大鼠脊髓背角BDNF的含量和蛋白表达随时间的变化。结果：（1）ELISA定量分析显示,脊神经结扎可以显著上调脊髓背角BDNF的含量。SNL后,背角BDNF的含量在24 h内即可由手术前的（91.09±7.1）pg/mg总蛋白上升到（160.75±15.0）pg/mg总蛋白（P〈0.01）,这种上调的高峰可以持续到SNL后的48 h;然而在假手术大鼠,背角BDNF的含量则没有明显的变化。与假手术组相比,SNL大鼠背角BDNF上调的高峰期在24~48 h（P〈0.01）,此后其含量开始逐渐恢复,至SNL后28 d基本恢复至术前对照水平（P〉0.05）;（2）免疫组化染色显示,在脊神经结扎的24h内,SNL大鼠结扎侧背角BDNF的蛋白表达也呈现明显的时间依赖性上调变化。SNL后12 h,背角浅层BDNF免疫反应的平均光密度值即可由手术前的0.18±0.01上调到0.25±0.03（P〈0.05）。与假手术组相比,在SNL后的12 h和24 h,SNL大鼠结扎侧背角浅层BDNF的免疫反应较假手术大鼠也明显上调,其平均光密度值分别由假手术大鼠的0.19±0.02和0.18±0.01上调到0.25±0.03（P〈0.05）和0.23±0.01（P〈0.05）。结论：脊神经结扎可以呈时间依赖性的上调脊髓背角BDNF的含量和蛋白表达,这种上调的BDNF可能在外周神经损伤所引起的神经病理痛的早期发挥作用。Objective ： sal horn in spinal nerve investigate the changes of BDNF content and immunoresponse in the spinal dorligated （SNL） rats and to explore the role of spinal BDNF in the development of neuropathic pain. Methods： Thirty male Sprague-Dawley rats were randomly divided into SNL-operated group and sham-operated group, and each group has fifteen rats. The changes of BDNF content and immunoresponse in the spinal dorsal horn after SNL- or sham-operated surgery were detected by enzymelinked immunosorbent assay （ELISA） and immunohistochemical staining techniques. Results ： （ 1 ） The BDNF content measured by ELISA was significantly increased after spinal nerve ligation. The BDNF content increased to （160.75 ± 15.0） pg/mg total protein from （91.09 ±7.1 ） pg/mg total protein of preoperation during 24 hours of SNL surgery （ P 〈 0.01 ）, and the maximal enhancement lasted for 48 hours. No significant changes in the BDNF content were observed in shame-operated rats. As compared with sham-operated rats, the significant enhancement of the BDNF content was observed at 24 - 48 h after surgery in SNL rats （P 〈0.01 ）, and then the content was decreased gradually. The BDNF content recovered to the level of pre-operation at 28 d after SNL （ P 〉 0. 05 ）. （2） The BDNF-immunoresponses （BDNF-ir） in the spinal dorsal horn were also increased significantly in a time-dependent manner during 24 hours of SNL. The mean IOD value of the BDNF-ir was increased to 0.25 ±0.03 from 0.18 0.01 of the pre-operation at 24 h of SNL （P 〈 0. 05）. As compared with sham-operated rats, the significant enhancement of the BDNF-ir was observed at 12 h （0.25 ± 0.03 versus 0.19± 0.02, P 〈 0. 05 ） and 24 h （0.23±0.01 versus 0.18 ± 0.01, P 〈 0.05）. Conclusion： Up-regulation of the BDNF content and immunoresponses in the spinal dorsal horn occurs in a time-dependent manner after spinal nerve ligation. The dorsal horn BDNF plays an important role in the development of neuropathic pain a

关 键 词：脑源性神经营养因子 脊髓背角 神经病理痛 免疫组化 ELISA 脊神经结扎 

分 类 号：R741[医药卫生—神经病学与精神病学]