不同途径注射含人脑啡肽原基因重组腺相关病毒对慢性神经病理痛大鼠的镇痛作用  

Analgetic effect mediated by recombinant adeno-accosiated virus with human preproenkephalin gene in different injection ways

作　　者：杨辉[1] 高峰[1] 安珂[1] 田学愎[1] 王鹏[1] 田玉科[1] 江辉[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学教研室,武汉430030

出　　处：《中华实验外科杂志》2013年第7期1493-1496,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872440）

摘　　要：目的构建含人前脑啡肽原基因的腺相关病毒载体（rAAV／hPPE），将此病毒载体通过不同途径注射至慢性疼痛模型大鼠体内，观察rAAV介导的hPPE体内转染的镇痛作用。方法将阳性对照质粒rAAV／hPPE分别经蛛网膜下、尾静脉和骨骼肌3种途径注射至慢性挤压伤疼痛模型大鼠体内，观察注射后大鼠热痛阈和机械痛阈的变化，8周后，处死大鼠，运用逆转录一聚合酶链反应（RT-PCR）方法测定大鼠组织hPPE的表达，用放射免疫方法测定大鼠器官组织中亮氨酸．脑啡肽（L—EK）的含量。结果行为学结果说明3种给药方式均可在5d-8周内对坐骨神经慢性挤压（CCI）热痛阈和机械痛阈显著升高；L—EK在脊髓组织中表达水平最高的是鞘内注射组而在血管和心肌组织中表达最高的是静脉注射组；PCR结果表明3种给药方式导致不同组织hPPEmRNA表达水平增高。结论将rAAV／hPPE注射到CCI大鼠体内，rAAV能将hPPE导入易感细胞，并获得较好的镇痛效果。Objective To observe the analgetic effect of recombinant adeno-accosiated virus with human preproenkephalin gene （rAAV/hPPE） on chronic constriction injury （CCI） rats in different injec- tion ways. Methods Constructed rAAV/hPPE was injected into rats by different routes： subarachnoid cav- ity, tail vein and skeletal muscle. The changes in paw withdrawal mechincal threshod （PWMT） and paw withdrawal thermal latency （PWTL） were observed for 8 weeks to evaluate the hyperalgesia. Two months after injection, reverse transcriptase-polymerase chain reaction （RT-PCR） and radioimmunoassay were used to detect the expression and secretion of hPPE in rats. Results The behavior test results indicated that all three ways of injection could increase the PWMT and PWTL from 5th day to 8th week in CCI rats. RT-PCR revealed that the three ways of injection all could increase the expression of hPPE mRNA in differ- ent organs. The expression level of Leucin-enkephalin （L-EK） in spinal cord was highest in the intrathecal injection group, and that in vein and myoeardium was highest in the tail vein injection group. Conclusion rAAV can transfect hPPE gene into neurons and NSCs with high efficiency, rAAV/hPPE injection can re- lease hyperalgesia in rat CCI models.

关 键 词：腺相关病毒 脑啡肽原 疼痛 

分 类 号：R74[医药卫生—神经病学与精神病学]