鞘内注射重组大鼠脂质运载蛋白-2对大鼠吗啡镇痛效能的影响  

The effect of intrathecal injection of recombinant rat lipocalin-2 on morphine analgesia efficacy in rats

作　　者：王芬[1] 谢滔[1] 江姿潞 刘清珍[1] 刘健[1] 李伟彦[1] 

机构地区：[1]南京军区南京总医院麻醉科,210002

出　　处：《临床麻醉学杂志》2015年第9期896-900,共5页Journal of Clinical Anesthesiology

基　　金：江苏省自然科学基金面上研究项目(BK20131328)

摘　　要：目的 观察鞘内注射重组大鼠脂质运载蛋白-2（LCN2）对大鼠吗啡镇痛效能的影响,并探讨其分子机制。方法 健康雄性SD大鼠32只,体重150~180g,采用随机数字表法将鞘内置管成功的大鼠随机分为四组（n=8）：Ⅰ组为对照组：鞘内注射MES缓冲液10μl,Ⅱ组、Ⅲ组和Ⅳ组分别鞘内注射10μl含LCN2蛋白0.02、0.2、2μg的MES溶液,每天1次,连续5d。分别在鞘内给药前和给药后第6、7、8天皮下注射吗啡10mg/kg,吗啡注射前和注射后45min测定大鼠热辐射缩足潜伏期（PWTL）,并计算最大可能镇痛效应百分比（MPE）。第8天行为学测试结束后处死动物,取脊髓腰膨大,采用Western blot法检测磷酸化p38丝裂原活化蛋白激酶（p-p38 MAPK）的表达;采用免疫组织化学法检测星型胶质细胞标志物胶质纤维酸性蛋白（GFAP）的表达。结果 与Ⅰ组和给药前比较,Ⅱ组大鼠鞘内给药后基础PWTL和MPE差异无统计学意义,Ⅲ组、Ⅳ组鞘内给药后第6、7、8天基础PWTL和MPE均明显降低（P〈0.05）;与Ⅰ组比较,Ⅱ组鞘内给药后脊髓腰膨大内pp38 MAPK、脊髓背角GFAP表达差异无统计学意义,Ⅲ组、Ⅳ组脊髓腰膨大内p-p38MAPK、脊髓背角GFAP表达明显增加（P〈0.05）。结论 大鼠连续5d鞘内注射LCN2蛋白0.2、2μg能够诱导热痛敏并导致吗啡镇痛效能下降,其机制可能与活化脊髓内星型胶质细胞和p38 MAPK有关。Objective To investigate the effect of intratheeal injection （IT） of recombinant rat lipocalin-2 （LCN2） on morphine analgesia efficacy in rats and the possible related molecular mecha- nism. Methods After successful intrathecal implantation with PE-10 catheter, male Sprague-Dawley rats weighing 150-180 grams were rando rely assigned to 4 groups （n=8） ： group I ： control group, IT MES solution 10 μl once a day for 5 consecutive days. Group Ⅱ-Ⅳ ： IT MES solution 10μl contai- ning LCN2 0. 02, 0.2, 2 μg once a day for 5 consecutive days respectively. Morphine （10 mg/kg） was subcutaneously injected before （Record as Day 0） and after （Day 6,7,8） the IT administration. Paw withdrawal latency to thermal stimuli （PWTL） was measured before morphine injection and 45 minutes after morphine injection. The percentage of maximal possible effect （MPE） was calculated later. Animals were sacrificed on the 8th day after behavioral test and the lumbar enlargement seg- ments of the spinal cord were removed for determination of the expression of phosphorylated-p38 mi- togen-activated protein kinase （p-p38 MAPK） by Western Blot analysis and glial fibrillary acid protein （GFAP） by immunohistochemical. Results Before IT, the basic PWTL and MPE of four groups had no significant difference. Compared to group I and before IT, after IT administration basic PWTL and MPE were significantly reduced while p-p38 MAPK and GFAP were markedly up-regulated in group Ⅲ and group Ⅳ （P〈0.05）. Conclusion IT administration of LCN2 0. 2 or 2 μg once a day for 5 consecutive days can induce thermal hyperalgesia and reduce morphine analgesic efficacy in rats pos- sibly through activating astrocytes and p38 MAPK in the spinal cord.

关 键 词：脂质运载蛋白-2 吗啡镇痛效能 脊髓 磷酸化P38丝裂原活化蛋白激酶 星型胶质细胞 

分 类 号：R614[医药卫生—麻醉学]