Gene gun particle encoding preproenkephalin cDNA produces analgesia against capsaicin-induced bladder pain in rats
Gene gun particle encoding preproenkephalin cDNA produces analgesia against capsaicin-induced bladder pain in rats
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DOI：
10.1016/j.urology.2004.10.070
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发表时间：
2005-04-01
期刊：
影响因子：
2.1
通讯作者：
Yoshimura, N
中科院分区：
文献类型：
--
作者：
Chuang, YC;Yang, LC;Yoshimura, N
目标。利用辣椒素诱导的大鼠膀胱痛模型，评价基因枪和直接注射基因治疗人前脑啡肽原(PPE)基因治疗大鼠膀胱疼痛的效果。阿片肽在调节排尿反射和控制炎性疼痛中起着重要作用。PPE就是这样一种前体分子。将人PPE基因克隆到pCMV载体中，通过直接注射或基因枪法将其导入成年雌性大鼠膀胱壁。基因治疗后第4天和第7天，在乌拉坦麻醉下进行连续膀胱测压，注入生理盐水(0.08mL/min)，然后注入15亩M辣椒素。用免疫组织化学方法检测PPE基因转移后膀胱内脑啡肽的表达。对照组和直接注射PPE基因组大鼠膀胱内注射辣椒素后收缩间期缩短(分别减少65.0%和63.1%)。然而，基因枪组在第4天和第7天对辣椒素滴注的反应显著降低(收缩间期分别减少16.2%和42.8%)。静脉注射阿片类拮抗剂纳洛酮(5 mg/kg)可逆转这种镇痛作用。基因枪组大鼠膀胱脑啡肽免疫反应在第4天增强，第7天减弱。利用基因枪的方法可以有效地将PPE基因转移到膀胱并抑制其伤害性反应。这些结果支持了PPE基因枪给药系统在治疗膀胱痛和其他类型内脏痛方面的潜在临床应用。泌尿外科65：804-810,2005。(C)2005年爱思唯尔公司。
Objectives. To evaluate the efficacy of gene therapy using a gene gun or direct injection for the transfer of human preproenkephalin (PPE) plasmid cDNA using a capsaicin-incluced bladder pain model in rats. Opioid peptides play an essential role in the modulation of micturition reflex and control of inflammatory pain. PPE is one such precursor molecule.Methods. Human PPE cDNA was cloned into a modified pCMV plasmid and delivered into the bladder wall of adult female rats by direct injection or gene gun. At 4 and 7 days after gene therapy, continuous cystometrograms were performed under urethane anesthesia by filling the bladder (0.08 mL/min) with saline, followed by 15 mu M capsaicin. Immunohistochemical staining was used to detect enkephalins in the bladder after PPE cDNA transfer.Results. The intercontraction interval was decreased after intravesical instillation of capsaicin (65.0% and 63.1% decrease) in the control group or direct PPE gene injection group, respectively. However, the gene gun-treated group showed a significantly reduced response to capsaicin instillation at day 4 and day 7 (intercontraction interval 16.2% and 42.8% decrease, respectively). This analgesic effect was reversed by intravenous naloxone, an opioid antagonist (5 mg/kg). Increased enkephalin immunoreactivity in the bladder was observed in the gene gun-treated group at day 4, which was reduced at day 7.Conclusions. The PPE gene can be effectively transferred and suppress the nociceptive response in the bladder using the gene gun method. These results support potential clinical application of PPE gene gun delivery system for the treatment of bladder pain and other types of visceral pain. UROLOGY 65:804-810, 2005. (c) 2005 Elsevier Inc.