Lu Ying, McNearney Terry A, Wilson Steven P, Yeomans David C, Westlund Karin N
Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX, USA.
Eur J Neurosci. 2008 Mar;27(5):1153-65. doi: 10.1111/j.1460-9568.2008.06076.x.
This study assessed enkephalin expression induced by intra-articular application of recombinant, enkephalin-encoding herpes virus (HSV-1) and the impact of expression on nociceptive behaviours and synovial lining inflammation in arthritic rats. Replication-conditional HSV-1 recombinant vectors with cDNA encoding preproenkephalin (HSV-ENK), or control transgene beta-galactosidase cDNA (HSV-beta-gal; control) were injected into knee joints with complete Freund's adjuvant (CFA). Joint temperatures, circumferences and nociceptive behaviours were monitored on days 0, 7, 14 and 21 post CFA and vector treatments. Lumbar (L4-6) dorsal root ganglia (DRG) and spinal cords were immunostained for met-enkephalin (met-ENK), beta-gal, HSV-1 proteins and Fos. Joint tissues were immunostained for met-ENK, HSV-1 proteins, and inflammatory mediators Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES) and cyclo-oxygenase-2, or stained with haematoxylin and eosin for histopathology. Compared to exuberant synovial hypertrophy and inflammatory cell infiltration seen in arthritic rats treated with CFA only or CFA and HSV-beta-gal, the CFA- and HSV-ENK-treated arthritic rats had: (i) striking preservation of synovial membrane cytoarchitecture with minimal inflammatory cell infiltrates; (ii) significantly improved nociceptive behavioural responses to mechanical and thermal stimuli; (iii) normalized Fos staining in lumbar dorsal horn; and (iv) significantly increased met-ENK staining in ipsilateral synovial tissue, lumbar DRG and spinal cord. The HSV-1 and transgene product expression were confined to ipsilateral lumbar DRG (HSV-1, met-ENK, beta-gal). Only transgene product (met-ENK and beta-gal) was seen in lumbar spinal cord sections. Targeted delivery of enkephalin-encoding HSV-1 vector generated safe, sustained opioid-induced analgesia with protective anti-inflammatory blunting in rat inflammatory arthritis.
本研究评估了关节内应用重组脑啡肽编码疱疹病毒（HSV-1）诱导的脑啡肽表达，以及该表达对关节炎大鼠伤害性反应行为和滑膜衬里炎症的影响。将携带编码前脑啡肽的cDNA的复制条件性HSV-1重组载体（HSV-ENK）或对照转基因β-半乳糖苷酶cDNA（HSV-β-gal；对照）注射到用完全弗氏佐剂（CFA）处理的膝关节中。在CFA和载体处理后的第0、7、14和21天监测关节温度、周长和伤害性反应行为。对腰段（L4-6）背根神经节（DRG）和脊髓进行免疫染色，检测甲硫氨酸脑啡肽（met-ENK）、β-半乳糖苷酶、HSV-1蛋白和Fos。对关节组织进行免疫染色，检测met-ENK、HSV-1蛋白以及炎症介质调节激活正常T细胞表达和分泌因子（RANTES）和环氧化酶-2，或用苏木精和伊红染色进行组织病理学检查。与仅用CFA处理或用CFA和HSV-β-gal处理的关节炎大鼠中出现的滑膜过度增生和炎性细胞浸润相比，用CFA和HSV-ENK处理的关节炎大鼠具有：（i）滑膜细胞膜结构显著保留，炎性细胞浸润极少；（ii）对机械和热刺激的伤害性反应行为显著改善；（iii）腰段背角Fos染色正常化；（iv）同侧滑膜组织、腰段DRG和脊髓中met-ENK染色显著增加。HSV-1和转基因产物表达局限于同侧腰段DRG（HSV-1、met-ENK、β-半乳糖苷酶）。在腰段脊髓切片中仅可见转基因产物（met-ENK和β-半乳糖苷酶）。靶向递送脑啡肽编码HSV-1载体在大鼠炎性关节炎中产生了安全、持续的阿片类药物诱导的镇痛作用，并具有保护性抗炎作用。
