鞘内注射TRESK-shRNA慢病毒对大鼠痛阈及脊髓JNK磷酸化的影响

发布时间：2018-02-06 05:51

  本文关键词： TRESK 背根神经节 脊髓 JNK　出处：《广东医学》2016年21期 　论文类型：期刊论文

【摘要】：目的评价鞘内注射TRESK-shRNA慢病毒对大鼠痛阈及脊髓JNK磷酸化的影响。方法健康雄性SD大鼠,体质量200~250 g,进行鞘内置管术。取鞘内置管成功的大鼠36只,采用随机数字表法分为3组(n=12):空白对照组(C组)、TRESK-shRNA慢病毒组(TRESK组)和阴性慢病毒组(V组)。TRESK组和V组分别鞘内注射TRESK-shRNA慢病毒和阴性慢病毒,10μL慢病毒,病毒滴度为1×10~8IFU/m L,1次/d,连续7 d;C组于相同时点鞘内注射10μL生理盐水。各组大鼠分别于鞘内注射前1 d(T0)及鞘内注射1 d(T1)、3 d(T2)、5 d(T3)、7 d(T4)测定机械刺激缩足阈(MWT)和热刺激缩足潜伏期(TWL)。鞘内注射7 d疼痛行为学测定结束后处死大鼠,取L4~5段背根神经节(DRG)和脊髓组织,realtime PCR检测背根神经节TRESK mRNA表达,Western blot检测脊髓JNK磷酸化水平。结果 TRESK组T3、T4时的MWT明显低于C组(P0.05)。与C组相比,TRESK组L4~5术侧DRG的TRESK mRNA表达明显降低(P0.05)。与C组相比,TRESK组L4~5脊髓的JNK磷酸化水平明显增高(P0.05)。结论鞘内注射TRESK-shRNA慢病毒可降低大鼠的机械痛阈,可能与激活脊髓JNK磷酸化有关。
[Abstract]:Objective to evaluate the effect of intrathecal injection of TRESK-shRNA lentivirus on pain threshold and JNK phosphorylation in spinal cord of rats. Thirty-six rats with successful intrathecal catheterization were divided into 3 groups by random digital table: control group C). TRESK-shRNA lentivirus group (TRESK group) and negative lentivirus group (group V). Group V and group V received intrathecal injection of TRESK-shRNA lentivirus and negative lentivirus, respectively. 10 渭 L lentivirus with a titer of 1 脳 10 ~ (8) IFU / m ~ (-1) per day for 7 days; In group C, 10 渭 L saline was injected intrathecally at the same time point. Rats in each group received intrathecal injection of T0 (1 day before intrathecal injection) and intrathecal injection of 1 day T1 (3 d) and 5 d (T3). After 7 days of intrathecal injection, the rats were killed after 7 days of intrathecal injection. The expression of TRESK mRNA in dorsal root ganglion (DRG) and spinal cord was detected by realtime PCR. Western blot was used to detect the level of JNK phosphorylation in spinal cord. Results the level of MWT in TRESK group was significantly lower than that in C group at T3 and T4, compared with that in C group. The expression of TRESK mRNA in DRG in TRESK group was significantly lower than that in group C (P 0.05). The level of JNK phosphorylation in TRESK group was significantly higher than that in the control group. Conclusion Intrathecal injection of TRESK-shRNA lentivirus can reduce the mechanical pain threshold of rats. It may be related to activation of JNK phosphorylation in the spinal cord.
【作者单位】： 广东省佛山市第一人民医院麻醉科;
【基金】：国家自然科学基金资助项目(编号:81300974) 广东省自然科学基金资助项目(编号:2015A030313899) 广东省医学科研基金资助项目(编号:A2015013)
【分类号】：R402
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本文编号：1493738