NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应  

Analgesic effect of NR2B peptide as subunit vaccine against neuropathic pain in rats

作　　者：冯颢[1] 王公明[1] 王红[1] 陈建平[1] 杨少兵[1] 许爱军[1] 曹菲[1] 田玉科[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉科,武汉市430030

出　　处：《中华麻醉学杂志》2008年第9期783-786,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金资助项目（30471660）

摘　　要：目的评价NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应。方法成年雌性SD大鼠，体重180～200g，制备保留性神经损伤（SNI）模型，7d后选取SNI模型制备成功的大鼠18只，随机分为3组（n=6），NR2B组沿背部两侧皮下多点注射NR2B多肽疫苗50μl（含NR2B100μg）3次，每隔2周注射1次；PBS组和KLH组分别给予PBS50μl和匙孔虫戚血蓝蛋白100μg，均用弗氏佐剂稀释至100μl。分别于制备SNI前1d（基础值）、第1次免疫前1d及第3次免疫后14d时测定机械痛阈；分别于制备SNI前1d及第3次免疫后14d时取血清和脑脊液，测定NR2B抗体滴度，然后处死大鼠，取L3-5，脊髓组织，其中3只采用免疫组化法测定脊髓背角胶质纤维酸性蛋白表达水平，反映星形胶质细胞激活程度；另外3只采用Western blotting法测定脊髓背角NR2B蛋白表达水平。结果NR2B组第3次免疫后14d时血清及脑脊液中均可检测到NR2B抗体，而SNI前1d时未检测到NR2B抗体，PBS组及KLH组各时点均未检测到NR2B抗体。与基础值比较，各组机械痛阈均降低（P〈0．05）；与PBS组和KLH组比较，NR2B组第3次免疫后机械痛阈升高，脊髓背角星形胶质细胞激活程度降低，NR2B蛋白表达下调（P〈0．05）。结论NR2B多肽疫苗对神经病理性痛大鼠具有镇痛作用，其机制与进入脊髓的NR2B抗体下调了NR2B蛋白表达有关。Objective To evaluate the analgesic effect of NR2B peptide as subunit vaccine against neuropathic pain in rats. Methods Healthy adult female SD rats weighing 180-200 g were used in this study. Spared nerve injury （SNI） was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve. Seven days after successful SNI surgery, 18 animals were randomly divided into 3 groups （ n = 6 each） ： group Ⅰ phosphate buffer solution （PBS）, group Ⅱ NR2B and group Ⅲ KLH. The animals in NR2B group was immunized with subcutaneous NR2B peptide vaccine 50 μl （containing NR2B peptide 100 μg） 3 times at 2 week intervals. Group PBS （50 μl） and KLH （ 100 μg） served as control groups. Threshold to noxious mechanical stimulation was measured 1 day before surgery （baseline）, 1 day before the 1st immunization and 14 days after the 3rd immunization using UGO mechanical pain threshold testing instrument （Baseline Co, Italy）. Blood and cerebral spinal fluid （CSF） samples were obtained 1 d before SNI surgrery （baseline） and 14 d after the 3rd immunization for detection of NR2B antibody. The animals were then killed and the lumbar segment （ L3- 5 ） of the spinal cord was removed for determination of expression of GFAP protein （by immuno-histochemistry） and NR2B protein （by Western blotting）. Results NR2B antibody was detected in serum and CSF at 14 d after the 3rd immunization in group NR2B, but not in group PBS and KLH at any interval. The threshold to mechanical stimulation was significantly decreased after SNI as compared with the baseline values in all 3 groups. The paw withdrawal threshold to mechanical stimulation after the 3rd immunization was significantly higher and the degree of activation of astrocytcs and the expression of NR2B protein in the dorsal horn of the spinal cord were significantly lower in NR2B group than in PBS and KLH groups. Conclusion The NR2B peptide vaccine can be used as a vaccine against

关 键 词：受体 N-甲基-D-天冬氨酸 疫苗 神经痛 

分 类 号：R741[医药卫生—神经病学与精神病学]