Tat-LK15运载NR2B siRNA治疗慢性炎性疼痛的效果  

The effects of NR2b siRNA carring vector Tat-LK15 on chronic pain

作　　者：杨雪[1] 吴昊鹏 彭捷[2] 陆建华[2] 

机构地区：[1]广州中医药大学第二附属医院广东省中医院麻醉科,广东广州510120 [2]广州军区广州总医院麻醉科,广东广州510010

出　　处：《广东医学》2017年第3期333-336,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81100817;81371233)

摘　　要：目的探讨大鼠鞘内注射细胞穿透肽Tat-LK15介导NR2B siRNA治疗大鼠慢性炎性疼痛的可行性。方法健康雄性SD大鼠72只,6周龄,体质量180~200 g,采用随机数字表法分为4组(n=18):空白对照组(C组)、慢性炎性痛模型组(CFA组)、Tat-LK15/NR2B siRNA组(si Tat-LK15组)和Tat-LK15/阴性对照siRNA组(nc Tat-LK15组)。除C组不给予任何处理外,其余大鼠右后趾皮下注射完全弗氏佐剂(CFA)制作慢性炎性痛模型。CFA组、si Tat-LK15组及nc Tat-LK15组于模型制备后第1、3、5天分别鞘内注射生理盐水、Tat-LK15/NR2B siRNA及Tat-LK15/阴性对照siRNA 10μL。每组取12只大鼠于造模前1 d及鞘内注射后3、7、14、21 d时测定机械缩足反应阈(PWMT)和热缩足反应潜伏期(PWTL)。每组取6只大鼠于鞘内注射后7 d处死后取脊髓,检测鞘内注射对大鼠脊髓NR2B蛋白表达的影响。结果鞘内注射后7 d,si Tat-LK15组大鼠脊髓NR2B蛋白表达下调。鞘内注射后3、7、14及21 d,si Tat-LK15组PWMT与PWTL与CFA组相比明显升高,差异有统计学意义(P<0.01),而nc Tat-LK15组PWMT与PWTL较CFA组相比差异无统计学意义(P>0.05)。结论鞘内注射Tat-LK15/NR2B siRNA可有效减轻CFA所致大鼠慢性炎性疼痛。Objective To discuss the effects of intrathecal injection of TAT- LK15 /NR2 B siRNA on chronic pain relieving in vivo. Methods Adult male SD rats were randomly divided into 4 groups,Group C（ control group,no treatment）,Group CFA（ inflammatory pain induced by freund＇s adjuvant complete and intrathecal injection of saline）,Group si Tat- LK15（ inflammatory pain model and intrathecal injection of Tat- LK15 / NR2 B siRNA） and Group nc Tat-LK15（ inflammatory pain model and intrathecal injection of Tat- LK15 / scrambled siRNA）. The changes of NR2 B expression were detected using western- blot in SCDH of chronic inflammatory pain rats following intrathecal injection. Pain control efficacy was evaluated by 50% mechanical withdrawal threshold（ PWMT） and thermal withdrawal latency（ PWTL）. Results NR2 B protein expression was efficiently inhibited by intrathecal injection of Tat- LK15 /NR2 B siRNA complexes compared with CFA group（ P〈0. 01）,while complexes injection of TAT- LK15 / scrambled siRNA did not show this inhibitory effect. Moreover,injection of Tat- LK15 / NR2 B siRNA complexes significantly increased PWMT and PWTL in chronic inflammatory rats compared with CFA group（ P〈0. 01）. Conclusion TAT- LK15 can efficiently deliver siRNA targeting NR2 B in vivo and relieve chronic inflammatory pain.

关 键 词：注射 鞘内 细胞穿透肽 基因疗法 转染 RNA 小分子干扰 受体 N-甲基-D-天冬氨酸 炎性痛 

分 类 号：R745.4[医药卫生—神经病学与精神病学]