CAJ | 학술논문

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-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hPPE mRNA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.
목적 구건함인전뇌배태원기인적선상관병독재체(rAAV/hPPE),장차병독재체통과불동도경주사지만성동통모형대서체내,관찰rAAV개도적hPPE체내전염적진통작용.방법 장양성대조질립rAAV/hPPE분별경주망막하、미정맥화골격기3충도경주사지만성제압상동통모형대서체내,관찰주사후대서열통역화궤계통역적변화,8주후,처사대서,운용역전록-취합매련반응(RT-PCR)방법측정대서조직hPPE적표체,용방사면역방법측정대서기관조직중량안산-뇌배태(L-EK)적함량.결과 행위학결과설명3충급약방식균가재5d～8주내대좌골신경만성제압(CCI)열통역화궤계통역현저승고；L-EK재척수조직중표체수평최고적시초내주사조이재혈관화심기조직중표체최고적시정맥주사조；PCR결과표명3충급약방식도치불동조직hPPE mRNA표체수평증고.결론 장rAAV/hPPE주사도CCI대서체내,rAAV능장hPPE도입역감세포,병획득교호적진통효과.
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 injection ways.Methods Constructed rAAV/hPPE was injected into rats by different routes:subarachnoid cavity,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 different organs.The expression level of Leucin-enkephalin (L-EK) in spinal cord was highest in the intrathecal injection group,and that in vein and myocardium 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 release hyperalgesia in rat CCI models.