国际麻醉学与复苏杂志   2014, Issue (9): 7-7
    
神经痛大鼠脊髓胶质细胞源性神经营养因子参与 多巴胺D2受体激动剂对痛觉过敏的调制作用
冷鑫, 隽立芹, 马正良, 夏小萍1()
1.徐州医学院
Neurotrophic factor derived from spinal cord glial cell line in rats with neuropathic pain involved in the modulation of dopamine D2 receptor agonists hyperalgesia
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摘要:

目的 研究鞘内注射多巴胺D2受体激动剂喹吡罗对坐骨神经慢性压迫损伤大鼠脊髓水平胶质细胞源性神经营养因子表达的影响,探讨其介导抗伤害作用的可能机制。 方法 实验1:采用完全随机分组方法将30只成年雄性SD大鼠制作坐骨神经慢性压迫损伤(chronic constriction injury of sciatic nerve, CCI)模型分为5组(每组6只):CCI+生理盐水(NS组)、CCI+喹吡罗0.1 μg(Q0.1组)、CCI+喹吡罗1 μg(Q1组)、CCI+喹吡罗5 μg(Q5组)、CCI+喹吡罗10 μg(Q10组),分别在建模后第7天鞘内注射0.1、1、5、10 μg喹吡罗,NS组单次鞘内注射生理盐水10 μl。 于给药前及给药后0.5、1、2、4、8、16 h测定大鼠术侧后足机械缩足反射阈值(paw withdrawal mechanical threshold, PWMT)和热缩足反射潜伏期(paw withdrawal thermal latency, PWTL)。实验2:将54只成年雄性SD大鼠制作CCI模型,并采用完全随机分组方法分为3组(Q5组、Q10组、NS组,每组18只):Q5组、Q10组分别在建模后第7天鞘内注射5、10 μg喹吡罗后0.5、1、2、4、8、16 h处死取材,NS组单次鞘内注射生理盐水10 μl;另取6只正常大鼠作为模型对照组(M组),另取6只正常大鼠作为对照组(C组)。采用免疫印迹法测定脊髓背角胶质细胞源性神经营养因子(glial cell line?蛳derived neurotrophic fact, GDNF)蛋白表达的变化。 结果 实验1: 与NS组比较,Q0.1组注药后各时间点PWMT和PWTL差异无统计学意义(P>0.05),Q1组注药后2 h PWMT [(4.3±1.5) g]及PWTL [(13.2±1.6) s],Q5组注药后1、2、4 h PWMT [(4.7±1.6)、(5.3±1.6)、(4.7±2.1) g]和PWTL[(14.0±1.7)、(15.2±1.5)、(13.4±1.6) s],Q10组注药后1、2、4 h PWMT [(6.0±1.3)、(7.3±1.0)、(5.3±2.1) g]和PWTL [(15.3±1.8)、(17.5±1.2)、(14.9±1.7) s]明显升高(P<0.05)。实验2:与C组比较,M组GDNF表达(0.95±0.09)明显升高(P<0.05)。与M组和NS组比较,GDNF的表达Q5组[(1.47±0.12)、(1.24±0.05)]、Q10组[(1.63±0.08)、(1.27±0.06)]明显增加(P<0.05),并且可以至少持续至注药后4 h。 结论 鞘内注射喹吡罗能够显著增加脊髓水平GDNF表达,同时伴有大鼠痛行为改善,GNDF的表达上调可能参与了多巴胺D2受体激动剂对神经病理性疼痛的调制过程。

关键词: 神经病理性疼痛; 多巴胺激动剂; 脊髓; 胶质细胞源性神经营养因子; 鞘内注射
Abstract:

Objective To study the effects of intrathecal injection of dopamine D2 receptor agonist quinpirole on expression of glial cell line-derived neurotrophic factor in the spinal cord in rats with chronic constrictive injury(CCI) and explore its possible mechanism of mediating antinociception. Methods In this study, models were established CCI in male Sprague-Dawley rats. The experiment was performed as 2 parts. In part one, 30 CCI rats were randomly divided into 5 groups(n=6):CCI+saline(group NS),CCI+quinpirole 0.1 μg(group Q0.1), CCI+quinpirole 1 μg(group Q1), CCI+quinpirole 5 μg (group Q5),CCI+quinpirole 10 μg (group Q10). The drugs were injected intrathecally on day 7 after CCI, respectively. Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured before and at 0.5,1,2,4,8 h and 16 h after intrathecal injection. In part two 54 CCI rats were randomly divided into 3 groups(group Q5, Q10 and NS, n=18). Group Q5 and Q10 were sacrificed at 0.5,1,2,4,8,16 h after intrathecal quinpirole 5,10 μg on day 7 after CCI. Group NS, a single intrathecal injection of saline 10 μl. Group M, another 6 normal rats as model control group. Another 6 normal rats as control group C. The expression of glial cell line-derived neurotrophic fact (GDNF) in the spinal cord was determined by Western blot. Results Part one: compared with group NS, the PWMT and PWTL of group Q0.1 at each time point after injection had no statistically significant difference (P>0.05). The PWMT and PWTL of 2 h of group Q1 [(4.3±1.5) g, (13.2±1.6) s].1,2,4 h of group Q5 [(4.7±1.6),(5.3±1.6),(4.7±2.1) g, (14.0±1.7),(15.2±1.5),(13.4±1.6) s] and Q10 [(6.0±1.3),(7.3±1.0),(5.3±2.1) g,(15.3±1.8),(17.5±1.2),(14.9±1.7) s] after drug administration were significantly promoted(P<0.05). Part two: compared with group C, the expression of GDNF was significantly increased in group M (0.95±0.09)(P<0.05). Compared with group M and NS, group Q5[(1.47±0.12),(1.24±0.05)] and Q10[(1.63±0.08),(1.27±0.06)] significantly increased the expression of GDNF(P<0.05), and it can last until 4 h after injection at least. Conclusions Intrathecal injection of quinpirole can significantly increase the level of GDNF expression in the spinal cord. At the same time, the pain behavior was improved. The upregulation of GNDF expression is involved in the modulation of dopamine D2 receptor agonist on neuropathic pain.

Key words: Neuropathic pain; Dopamine agonist; Spinal; Glial cell line?蛳derived neurotrophic fact; Intrathecal injection