论文部分内容阅读
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: To investigate the changes of BDNF content and protein expression in spinal dorsal horn in spinal cord ligation (SNL) rats and to explore its role in the development of neuropathic pain. Methods: According to the principle of double-blind randomization, 30 male SD rats were randomly divided into SNL operation group and sham operation control group, 15 rats in each group. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry , To detect neuropathic pain rats spinal cord dorsal BDNF content and protein expression over time. Results: (1) ELISA quantitative analysis showed that ligation of spinal nerves significantly increased the content of BDNF in spinal dorsal horn. After SNL, the content of BDNF in the dorsal horn increased from (91.09 ± 7.1) pg / mg total protein before surgery to (160.75 ± 15.0) pg / mg total protein (P <0.01) within 24 h Can continue to 48 h after SNL; however, sham operation rats, BDNF content in the dorsal horn did not change significantly. Compared with the sham-operation group, the peak of BDNF upregulation in the dorsal horn of SNL rats was 24-48 h (P <0.01), and the content of BDNF began to recover gradually from the beginning to the 28th day after SNL (P> 0.05). (2) Immunohistochemical staining showed that BDNF protein expression in the ligation dorsal horn of SNL rats also showed a significant time-dependent up-regulation in 24h of ligation of spinal nerves. At 12 h after SNL, the mean optical density of BDNF immunoreactivity in the dorsal horn was increased from 0.18 ± 0.01 before surgery to 0.25 ± 0.03 (P <0.05). Compared with the sham group, the immunoreactivity of BDNF in the superficial layers of dorsal horn in SNL rats was significantly increased at 12 h and 24 h after SNL compared with that in sham-operated rats, and the average optical density values of sham-operated rats 0.19 ± 0.02 and 0.18 ± 0.01 to 0.25 ± 0.03 (P <0.05) and 0.23 ± 0.01 (P <0.05) respectively. Conclusion: Ligation of spinal nerves can up-regulate the content and protein expression of BDNF in spinal dorsal horn in a time-dependent manner. This upregulation of BDNF may play an important role in the early stage of neuropathic pain induced by peripheral nerve injury.