鞘内注射入β-内啡肽基因重组腺病毒对慢性神经病理痛大鼠的镇痛作用  

Analgesic effect of intrathecal recombinant adenovirus containing human beta-endophin gene in a rat model of neuropathic pain

作　　者：尤圣武[1] 徐学武[1] 俞卫锋[1] 于布为[2] 钱其军[3] 苏长青[3] 王星华[3] 

机构地区：[1]第二军医大学附属东方肝胆外科医院麻醉科,上海市200438 [2]上海第二医科大学瑞金医院麻醉科 [3]第二军医大学东方肝胆外科医院肿瘤病毒基因治疗研究室

出　　处：《中华麻醉学杂志》2005年第6期428-432,共5页Chinese Journal of Anesthesiology

摘　　要：目的观察鞘内注射入β-内啡肽(β-EP)基因重组腺病毒(Ad-NEP)对慢性神经病理痛大鼠的镇痛作用。方法36只雄性SD大鼠,体重210-260 g,随机分为手术组(n=26)、假手术组(n= 5)、空白对照组(n=5),手术组分为三个亚组:Ad-NEP组(n=9)、绿色荧光蛋白基因重组腺病毒(Ad- GFP)组(n=9)、生理盐水组(n=8)。大鼠腹腔注射氯胺酮100 mg/kg和阿托品50 mg/kg麻醉后,手术组构建大鼠坐骨神经慢性捆扎(CCI)模型,假手术组进行同样手术,但不捆扎坐骨神经,空白对照组不进行手术;手术组经L5,6间隙蛛网膜下腔置入PE-10导管,7d后分别注射1×108pfu的Ad-NEP、Ad- GFP和40μl生理盐水。于手术后7 d(T0)、鞘内注射前当日(T1)、注射后1 d(T2)、1周(T3)、2周(T4)、3 周(T5)、4周(T6)、5周(T7)分别测定五组大鼠的左右足热痛阈;T3时取Ad-NEP、Ad-GFP组大鼠各1 只,取脊髓L3-6段作免疫组化检测;鞘内注射后10 d取Ad-NEP、Ad-GFP组大鼠,经腹腔内注射1 mg/kg 纳络酮,每间隔10min记录右足热痛阈(t0-9,共观察90min),并分别测定这两组T1-7时的脑脊液内β- EP浓度。结果Ad-GFP组和生理盐水组大鼠右足热痛阈明显低于假手术组和空白对照组(P< 0.01)。Ad-NEP组在T0,1,7时右足热痛阈明显低于假手术组和空白对照组(P<0.01),在T2-6时的右足热痛阈高于T0,也高于Ad-GFP组和生理盐水组(P<0.05或0.01),t1-6时右足热痛阈低于t0时(P< 0.01),与Ad-GFP组和生理盐水组比较差异无统计学意义(P>0.05)。Ad-NEP组脊髓外膜可见明显的橙黄色浓染带,后角区可见少量橙黄色细胞,T2-7时脑脊液内β-EP浓度均高于Ad-GFP组(P< 0.01。)结论鞘内注射重组腺病毒Ad-NEP对慢性神经病理痛大鼠有明显的镇痛效果,作用时间长达4周以上。Objective To examine the analgesic effect of intrathecal （i. t. ） adenovirus containing humanbeta-endorphin （β-EP） gene in a rat model of neuropathic pain produced by chronic constrictive injury （CCI）.Methods Thirty-six male SD rats weighing 210-260 g were randomly divided into 3 groups： （1） blank controlgroup （ n = 5） ; （2） sham-operated group （ n = 5） ; （3） neuropathic pain group （ n = 26）. The neuropathic pain group was further divided into 3 subgroups： Ad-NEP subgrouop （n = 9）; Ad-GFP （the recombinant adenovirus containing green fluorescent protein） subgroup （n = 9） and normal saline （NS） subgroup （n = 8）. The animals were anesthetized with intraperitoneal ketamine 100 mg · kg^-1 and atropine 50 mg · kg^-1 . Neuropathic pain was produced by ligation of right sciatic nerve according to the technique described by Bennet and Xie. In shamoperated group the sciatic nerve was exposed but not ligated. In blank control group no operation was performed.Seven days after the surgery a PE-10 catheter was placed in the subarachnoid space at L5.6 according to the method of Milligan et al. Seven days after catheter placement 1 × 10^8 pfu Ad-NEP, Ad-GFP and 0.9% saline were injected i.t. via the catheter respectively. The paw-withdrawal latency to radiant heat was measured before surgery（T0,baseline）, the day of i.t. injection （T1） and 1 day （T2）, 1 w （T3）, 2 w （T4）, 3 w （T5）, 4 w （T6） 5 w（T7） after i.t. injection. At one week after i.t. administration one animal in Ad-NEP subgroup and Ad-GFP subgroup was killed and the lumbar segment （ L3-6） of the spinal cord was removed for immuno-histochemical examination. Naloxone 1 mg·kg^-1 was given intraperitoneally in Ad-NEP subgroup （n = 8） and Ad-GFP subgroup（ n = 8） at 10 days after i.t. injection. Pain threshold to thermal stimulation of the right paw was measured before（t0） and from 10-90 min after intraperitoneal naloxone injection at an interval of 10 min （t1-9�

关 键 词：鞘内注射 人β-内啡肽 基因重组腺病毒 慢性神经病理痛 镇痛 

分 类 号：R96[医药卫生—药理学]