| 摘要: |
| 目的通过观察3型酸敏感离子通道（ASIC3）信号通路阻断剂阿米洛利对髓核致炎大鼠背根神经节（DRG）中ASIC3表达的影响，探索ASIC3在髓核组织诱导发生根性疼痛过程中的可能机制。方法48只大鼠采用随机数字表法分为对照组、模型组和阿米洛利组3组，每组16只。对照组在暴露L5DRG后直接缝合。模型组取出大鼠尾部的髓核组织置于暴露的L5DRG上，缝合切口。阿米洛利组在模型组的基础上，于术后2周内每天腹腔内注射20滋g/kg阿米洛利。对照组和模型组不进行药物干预。造模成功后分别于术后2、4、6、8、10、12d检测大鼠后足的机械刺激缩足阈值（PWT）。采用RT-PCR法和Westernblot法测定ASIC3、TNF-α和IL-6mRNA和蛋白表达水平。术后1、3、7和14d采用ELISA法检测外周血及DRG上清液中的TNF-α和IL-6水平，并检测一氧化氮（NO）水平。结果与对照组比较，术后12d时模型组大鼠后足的PWT显著下降（P＜0.01），术后14d模型组大鼠DRG中ASIC3的mRNA和蛋白水平上调（均P＜0.01），外周血和DRG中NO、TNF-α、IL-6水平均上调（均P＜0.01）。与模型组比较，阿米洛利组大鼠后足的PWT在8d内下降，之后逐渐增加，术后10和12d大鼠后足的PWT明显高于模型组（均P＜0.01），术后14dDRG中ASIC3的mRNA和蛋白水平下降（均P＜0.05），外周血和DRG中NO、TNF-α、IL-6水平均显著降低（均P＜0.01）。结论ASIC3可能在由髓核突出引起的神经根性疼痛中起重要作用。ASIC3可能通过上调NO及细胞炎性因子（TNF-α和IL-6）的表达并增强这些介质的疼痛加强作用，诱导痛觉过敏从而引起神经根性疼痛。 |
| 关键词: 椎间盘 髓核 3 型酸敏感离子通道 炎症 |
| DOI：10.12056/j.issn.1006-2785.2017.39.24.2017-918 |
| 分类号: |
| 基金项目:浙江省中医药科技计划项目（2015ZA157）；杭州市科技计划重点项目（20150733Q60） |
|
| Effect of amiloride on ASIC3 expression in dorsal root ganglion following nerve root inflammation induced by nucleus pulposus in rats |
|
|
|
Guangxing Hosptial Affiliated to Zhejiang Medicine University
|
| Abstract: |
| Objective To investigate the effect of amiloride on the expression of acid sensing ion channel 3 (ASIC3) in dorsal root ganglion (DRG) following nerve root inflammation induced by nucleus pulposus in rats. Methods Forty eight SD rats were randomly divided into control group(Sham, n=16), inflammation group (NP, n=16), inflammation+amiloride group (NP+Ami, n=16). The L5 DRG was exposed in rats of NP and NP+Ami groups by surgery, and sham surgery was performed in sham group. Intraperitoneal injection of amiloride 20滋g/kg was given to rats in NP+Ami group q.d for 2 weeks after surgery, and no amiloride was given to rats in sham group and NP group．The mechanical pain withdrawal threshold (PWT) was evaluated at 2, 4, 6, 8, 10 and 12d after modeling. The expression of ASIC3, TNF- α, IL-6 mRNA and protein were examined by Real-time PCR and Western blot, respectively. TNF-α and IL-6 concentrations present in peripheral blood and DRG were determined at 1, 3, 7 and
14d after the surgical procedures using ELISA method. NO levels were examined by biochemistry test. Results Rats exposed to the NP showed decreased PWT at 12d after surgery (P ＜0.01), and the levels of ASIC3 and the NO, TNF- α, IL-6 were up-regulated in DRG induced by NP 14d after surgery(all P＜0.01). Compared to NP group the mechanical PWT decreased at 8d after surgery and then gradually increased at 10 and 12d after surgery in NP+Ami group (all P ＜0.01). ASIC3 was down-regulated (both P＜0.05), and the levels of NO, TNF-α, IL-6 were significantly decreased at 14d after surgery (all P＜ 0.01) in NP+Ami group. Conclusion ASIC3 may upgrade the expression of NO and inflammatory cytokines TNF-α and IL-6 to
induce hyperalgesia and never root pain. |
| Key words: Intervertebral disc Nucleus pulposus ASIC3 Inflammation |
