|
| TMEM16A下调缓解坐骨神经分支选择性损伤大鼠的神经病理性痛 |
| Down-regulation of TMEM16A relieves neuropathic pain in sciatic nerve injury rats |
| |
| DOI：10.12089/jca.2019.09.017 |
| 中文关键词: TMEM16A 神经病理性痛 背根神经节 痛觉过敏 |
| 英文关键词: TMEM16A Neuropathic pain Dorsal root ganglion Hyperalgesia |
| 基金项目:国家级大学生创新创业训练计划项目(201810759310) |
|
| 摘要点击次数: 2877 |
| 全文下载次数: 753 |
| 中文摘要: |
目的 探讨TMEM16A在坐骨神经分支选择性损伤(SNI)神经病理性痛模型大鼠中的作用。
方法 成年雄性SD大鼠,体重180～220 g,随机分为假手术组(sham组,n=6)和SNI组(n=12),sham组大鼠仅暴露左侧坐骨神经分支;SNI组行SNI。在术前1 d、术后3、7、10、14 d测定大鼠热缩足潜伏期(TWL)、冷缩足潜伏期(CWL)和机械缩足阈值(MWT)。采用Western blot测定术前1 d和术后7、14 d术侧背根神经节(DRG)中TMEM16A蛋白含量。另取72只雄性SD大鼠随机分为四组: sham+生理盐水组(CS组)、SNI+生理盐水组(SS组)、SNI+CaCCinh-A01组(SC组)和SNI+T16Ainh-A01组(ST组)，每组18只。在给药前3 d行鞘内置管术,CS、SS组大鼠术后14 d鞘内单次注射生理盐水10 μl,SC、ST组大鼠相同时点鞘内单次注射10 μl浓度为1 mg/ml的特异性钙激活氯通道(CaCCs)抑制剂CaCCinh-A01或T16Ainh-A01,在给药后的8 h内每隔1 h测定TWL、CWL和MWT。另设相同四组大鼠于术后第12天开始每隔6 h分别鞘内注射10 μl的生理盐水、CaCCinh-A01或T16Ainh-A01,共注射5次,于术后第14天提取术侧DRG进行Western blot和免疫荧光实验,观察TMEM16A蛋白含量及TMEM16A分布特点。
结果 与sham组比较,SNI组术后3、7、10、14 d CWL明显延长(P<0.05),MWT明显降低(P<0.05),TWL差异无统计学意义。与术前1 d比较,SNI组TMEM16A蛋白含量在术后7、14 d明显增高,且术后14 d明显高于术后7 d(P<0.05)。与SS组比较,SC组和ST组CWL从给药后1 h开始降低,3 h达到最低，且在给药后的1～4 h内SC组CWL明显小于ST组(P<0.05);MWT从给药后1 h开始升高,分别在2 h和3 h达到最高且在给药后的1、2、4、7和8 h内SC组MWT明显高于ST组(P<0.05);TWL在各时点差异均无统计学意义。与SS组比较,SC组和ST组TMEM16A蛋白含量明显降低(P<0.05),且ST组明显低于SC组(P<0.05)。免疫荧光结果显示TMEM16A主要表达在与伤害感受相关的中小神经元上。
结论 TMEM16A可能在SNI诱导的持续性痛觉过敏中起关键作用。TMEM16A可为神经病理性痛提供新的药物靶点。 |
| 英文摘要: |
Objective To evaluate the role of TMEM16A in neuropathic pain elicited by sciatic nerve injury (SNI).
Methods Adult male Sprague-Dawley rats weighing 180-220 g were randomly divided into sham-operated group (group sham, n = 6) and group SNI (n = 12). Sham-operated rats only exposed left sciatic nerve branches; group SNI underwent sciatic nerve injury. Rat behavioral measurements were performed 1 day before surgery and 3, 7, 10, 14 days after surgery including thermal withdrawal latency (TWL), cold withdrawal latency (CWL), and mechanical withdrawal threshold (MWT). The expression of TMEM16A protein in DRG was measured by Western blot on the 1st day before operation and on the 7th and 14th day after surgery. Another 72 male Sprague-Dawley rats were randomly divided into four groups (n = 18): sham+saline group (group CS), SNI+saline group (group SS), SNI+CaCCinh-A01 group (group SC) and SNI+T16Ainh-A01 group (group ST). Intrathecal catheterization was performed 3 days before administration. The groups CS and SS received a single intrathecal injection of 10 μl of normal saline on the 14th day after surgery. In the groups SC and ST, a single intrathecal injection of 10 μl of specific CaCCs inhibitor CaCCinh-A01 or T16Ainh-A01 at a concentration of 1 mg/ml was performed. The rats underwent behavior measurements every 1 h within 8 h after administration. In the same four groups of rats, 10 μl of normal saline, CaCCinh-A01 or T16Ainh-A01 were injected intrathecally every 6 hours from the 12th day after surgery with total 5 times. The DRG was extracted on the 14th day after operation. Western blot and immunofluorescence experiments were performed to observe the changes of TMEM16A protein content and the temporal and spatial distribution of TMEM16A.
Results Compared with the group sham, CWL was significantly increased (P < 0.05), MWT was significantly decreased (P < 0.05), and TWL was not significantly different 3, 7, 10, and 14 days after surgery in the group SNI. Compared with 1 day before surgery, the protein content of TMEM16A in DRG of group SNI increased 7 days after operation, and it was significantly higher 14 days after surgery than 7 days after surgery (P < 0.05). Compared with the group SS, the CWL of the group SC and the group ST decreased from 1 h after administration, and reached the lowest at 3 h. The CWL of the group SC was significantly shorter than that of the group ST within 1-4 h after administration (P < 0.05). The MWT was increased from 1 h after administration and reached the highest at 2 h and 3 h, respectively. The MWT of the group SC was significantly higher than that of the group ST at 1, 2, 4, 7 and 8 h after administration (P < 0.05). There was no statistically significant difference in TWL at each time point. Compared with group SS, the protein contents of TMEM16A in group SC and group ST were significantly lower (P < 0.05), and it was significantly lower in group ST than in group SC (P < 0.05). Immunofluorescence results showed that TMEM16A was mainly expressed in small neurons associated with nociception.
Conclusion TMEM16A may play a key role in SNI-induced persistent hyperalgesia. TMEM16A provides a new drug target for neuropathic pain. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
