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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(P<0.05)。与C组相比,TRESK组L4~5术侧DRG的TRESK mRNA表达明显降低(P<0.05)。与C组相比,TRESK组L4~5脊髓的JNK磷酸化水平明显增高(P<0.05)。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。
Objective To evaluate the effect of intrathecal injection of TRESK-shRNA lentivirus on pain threshold and JNK phosphorylation in spinal cord of rats. Methods Healthy male Sprague-Dawley rats, weighing 200-250 g, were treated with intrathecal catheterization. Totally 36 rats were randomly divided into 3 groups (n = 12): blank control group (C group), TRESK-shRNA lentivirus group (TRESK group) and negative lentivirus group Group V). TRESK group and group V were intrathecal injection of TRESK-shRNA lentivirus and negative lentivirus, 10μL lentivirus, the virus titer of 1 × 10 ~ 8IFU / m L, 1 times / d, for 7 days; C group at the same time Intrathecal injection of 10 μL saline. Rats in each group were subjected to mechanical stimulation of contraction threshold (MWT) at 1 day before intrathecal injection (T0) and 1 d after intrathecal injection (T1), 3 days (T2), 5 days (T3) and 7 days ) And thermal stimulation contracting latency (TWL). After intrathecal injection for 7 days, the rats were sacrificed and the DRGs and spinal cord tissues from L4 to L5 were taken. Real-time PCR was used to detect TRESK mRNA expression in dorsal root ganglion. Western blot was used to detect the level of JNK phosphorylation . Results The MWT at T3 and T4 in TRESK group was significantly lower than that in C group (P <0.05). Compared with group C, the expression of TRESK mRNA in DRG of L4 ~ 5 in TRESK group was significantly decreased (P <0.05). Compared with group C, JNK phosphorylation of L4 ~ 5 spinal cord in TRESK group was significantly increased (P <0.05). Conclusion Intrathecal injection of TRESK-shRNA lentivirus can reduce the mechanical pain threshold in rats, which may be related to activation of spinal cord JNK phosphorylation.