国际麻醉学与复苏杂志   2014, Issue (8): 4-4
    
鞘内注入布比卡因对孕鼠神经功能及背根神经节天冬氨酸特异性半胱氨酸蛋白酶-9 mRNA的影响
崔睿, 徐世元, 胡微, 蔡清香, 张鸿飞1()
1.深圳市妇幼保健院
Effect of intrathecal bupivacaine on sensor motor function and on expression of cysteinyl aspartate specific protease-9 in dorsal root ganglion in pregnant rats
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摘要：

目的　研究鞘内注入布比卡因对孕鼠神经功能的影响，脊神经细胞的凋亡及其信号转导机制。　方法　取鞘内置管成功孕鼠18只，按完全随机分组方法分成3组（每组6只），孕鼠对照组（PN组）、2％布比卡因组（P2B组）与4％布比卡因组（P4B组）。另取鞘内置管成功非孕鼠18只，按完全随机方法分成3组（每组6只）：非孕鼠对照组（N组）、2％布比卡因组（2B组）与4％布比卡因组（4B组）。PN组与N组：鞘内注射生理盐水30 μl，其余4组相应鞘内分别注入2％或4％布比卡因30 μl。鞘内注药后不同时间点测定甩尾反应潜伏期（tail-flick latency， TFL），用最大效应百分比（maximal possible effect， MPE）表示，并进行下肢运动功能评分（motor function， MF）。4 d后各组大鼠行断颈处死，分离背根神经节，逆转录?蛳聚合酶链反应（reverse transcriptive polymerase chain reaction， RT-PCR）检测天冬氨酸特异性半胱氨酸蛋白酶（cysteinyl aspartate specific protease， caspase）-8和caspase-9 mRNA的表达。　结果　注药后MPE随时间发生显著变化（P<0.05），而且分组与时间之间存在交互作用（P<0.05），10 min~2 h之间，2B组、4B组、P2B组和P4B组MPE均高于N组和PN组（P<0.05），在1 h时点，P2B组和P4B组MPE均高于N组、PN组、2B组和4B组（P<0.05）；在10 min~4 h 之间2B组、4B组、P2B组和P4B组MF评分较N组与PN组均明显升高；与N组（851±201）和PN组（755±289）比较，2B组（3 852±1 247），4B组（4 273±1 513），P2B组（4 507±1 437）和P4B组（5 147±1 780） caspase?蛳9 mRNA表达均上调（P<0.05）。　结论　妊娠期间孕鼠背根神经节细胞对布比卡因的阻滞效能敏感性增大。鞘内注入相同浓度和剂量布比卡因，孕鼠感觉、运动神经阻滞持续时间明显长于非孕鼠。鞘内注入布比卡因会导致大鼠背根神经节内caspase?蛳9 mRNA表达上调。
关键词： 布比卡因；　妊娠；　神经毒性；　天冬氨酸特异性半胱氨酸蛋白酶-9；　凋亡
Abstract:

Ｏｂｊｅｃｔｉｖｅ　To investigate the effect of intrathecal bupivacaine on sensor motor function and on expression of caspase-9 and caspase-8 in dorsal root ganglion in pregnant or nonpregnant rats.　Methods　Eighteen female SD rats weighting 200 g-220 g and eighteen late pregnant rats weighting 350 g-400 g were used in this study. All the pregnant rats in which PE-10 catheter were successfully placed without complication were divided into 3 groups randomly（n=6）：normal pregnant control group （group PN）， 2％ bupivacaine group（group P2B） and 4％ bupivacaine group（group P4B）. All the nonpregnant rats in which PE-10 catheter were successfully placed without complication were divided into 3 groups randomly（n=6）， normal nonpregnant-control group （group N）， 2％ bupivacaine group （group 2B） and 4％ bupivacaine group（group 4B）. The rats of group PN and group N received normal saline 30 μl intrathecal. The rats of group P2B and group 2B received 2％ bupivacaine 30 μl intrathecal. The rats of group P4B and group 4B received 4％ bupivacaine 30 μl intrathecal Tail-flick test and motor function were performed at 10， 20， 30 min， 1， 2， 4 h， 1， 2， 3 d and 4 d after intrathecal administration. TF latencies were converted to the percent of maximal possible effect （MPE）. Four days later， the animals were sacrificed and the dorsal root ganglion were removed for determination of mRNA using reverse transcriptive polymerase chain reaction（RT-PCR）.　Results　MPE were different among the groups according timepoints after intrathecal. Among in different groups have ainteraction. MPE in group P2B and P4B were significantly higher compared with those in group 2B and 4B after intrathecal 2， 4 h and 1 d. MF scores were significantly different after intrathecal 10 min-4 h in rats， group 2B， 4B， P2B and P4B motor block time was longer than group N and PN. Rats in group 2B， 4B， P2B and P4B showed significantly increase the expression of caspase-9 mRNA in dorsal root ganglion compared with those in group N and PN.　Conclusions　Intrathecal injection of the same concentration and dosage of bupivacaine， pregnant rats sensory， motor nerve block lasted significantly longer than nonpregnant rats. The result indicated that the sensitivity of bupivacaine increased in pregnancy rats. Bupivacaine showed significantly increase the expression of caspase-9 mRNA in dorsal root ganglion cells.
Key words: Bupivacaine；　Pregnancy；　Neurotoxicity；　Cysteinyl aspartate specific protease-9；　Apoptosis