脉冲射频大鼠CCI模型背根神经节对脊髓小胶质细胞IRF8表达的影响

发布时间：2018-01-27 04:36

  本文关键词： 神经病理性疼痛 脉冲射频 背根神经节 小胶质细胞 IRF8 反义寡核苷酸　出处：《福建医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：【目的】本研究选用结扎大鼠一侧坐骨神经主干制备慢性坐骨神经缩窄损伤(CCI)模型,通过一般行为学、50%机械刺激缩爪反应阈值(50%MWT)、热辐射刺激缩爪反应潜伏期(TWL)观察判断对大鼠背根神经节(DRG)进行脉冲射频(PRF)治疗及鞘内注射干扰素调节因子8(IRF8)反义寡核苷酸是否具有缓解疼痛的作用,并通过Western-Blot、免疫组化(IHC)法检测大鼠脊髓组织中IRF8、离子钙接合蛋白Iba1表达来阐述脉冲射频背根神经节缓解疼痛的可能机制。【方法】第一部分脉冲射频大鼠CCI模型背根神经节对痛阈和脊髓IRF8表达的影响采用大鼠CCI模型,于造模后第7天行背根神经节脉冲射频治疗。造模前,造模后1、3、5、7天,脉冲射频后1、7、14天,通过观察一般行为学,50%MWT、TWL检测大鼠痛觉改变。造模后7天,脉冲射频后7、14天,Western-Bolt、IHC法检测IRF8蛋白水平。第二部分鞘内注射IRF8反义ODN对大鼠CCI模型痛阈和脊髓IRF8、Iba1表达的影响建立鞘内置管和CCI模型,于造模后12h分别进行鞘内注射IRF8反义ODN、IRF8错义ODN、等体积NS,每天1次,共6天。造模前,造模后1、3、5、7、10、12、14天大鼠实施一般行为学、50%MWT,TWL测定。造模后第7、14天采用Western-Blot法检测IRF8、Iba1蛋白表达。【结果】1、与Sham组比,CCI组、CCI+PRF组、CCI+NPRF组大鼠于神经损伤后右后爪50%MWT和TWL均显著降低(P0.01),同时脊髓IRF8蛋白表达上调(P0.05)。脉冲射频后1、7、14天CCI+PRF组与治疗前比较50%MWT和TWL升高(P0.01),并显著高于CCI组和CCI+NPRF组(P0.01),但仍低于Sham组(P0.01),脊髓IRF8蛋白的表达则相应减少(P0.05)。2、与Sham组比较,NS组、MM组大鼠神经损伤后右后爪50%MWT和TWL均显著降低(P0.01),脊髓IRF8、Iba1蛋白表达则增强(P0.05)。AS组与Sham组相比于CCI后第10天起右后爪50%MWT和TWL下降(P0.01),但仍明显高于MM组、NS组(P0.01)。CCI后第7、14天IRF8、Iba1蛋白表达,AS组较MM组、NS组显著减弱(P0.05),但仍强于Sham组(P0.05)。AS组第14天IRF8、Iba1蛋白较第7天增强(P0.05)。【结论】1、PRF可以减轻CCI模型痛敏,其机制可能是PRF抑制大鼠脊髓IRF8的表达,从而缓解疼痛。2、IRF8是诱导外周神经损伤后痛觉敏化的关键因子,提前给予阻断IRF8可以缓解神经病理性疼痛。
[Abstract]:[Objective] this study selected one side of sciatic nerve trunk for ligation of rat chronic constriction injury of the sciatic nerve (CCI) model, the general behavior of 50% mechanical stimulation, paw withdrawal threshold (50%MWT), thermal radiation to stimulate the paw withdrawal latency (TWL) in rat dorsal root ganglion (DRG) were observed pulsed radiofrequency (PRF) therapy and intrathecal injection of interferon regulatory factor 8 (IRF8) or antisense oligonucleotides has the effect of pain relief, and by Western-Blot, immunohistochemistry (IHC) method for the detection of IRF8 in spinal cord of rats, calcium binding protein Iba1 expression to explain the possible mechanism of RF pulse to relieve the pain of dorsal root ganglion [method] the first part. Effects of pulsed radiofrequency CCI model of rat dorsal root ganglion and spinal IRF8 expression on the pain threshold of the rat CCI model was used to model seventh days after dorsal root ganglion pulsed radio frequency treatment. Before modeling, modeling After 1,3,5,7 days, 1,7,14 days after the observation of pulsed radiofrequency, the general behavior, 50%MWT, TWL in rats were detected pain. 7 days after modeling, 7,14 days after pulsed RF Western-Bolt, detection of IRF8 protein level by IHC. The second part of intrathecal injection of IRF8 antisense ODN on CCI rat model of pain threshold and spinal cord IRF8, Iba1 expression and the effects of intrathecal catheter and CCI model, after modeling 12h respectively by intrathecal injection of IRF8 antisense ODN, IRF8 missense ODN, volume NS, 1 times a day, a total of 6 days. Before modeling, after modeling 1,3,5,7,10,12,14 rats in the implementation of the general behavior, 50%MWT, TWL determination of the first 7,14 days after modeling. Western-Blot method was used to detect IRF8 and Iba1 protein expression. [results] 1, compared with the Sham group, CCI group, CCI+PRF group, CCI+NPRF group of rats after nerve injury in the right hind paw of 50%MWT and TWL were significantly decreased (P0.01), while the expression of IRF8 protein in the spinal cord adjustment (P0.05). Pulsed RF 1,7,14 days after CCI +PRF缁勪笌娌荤枟鍓嶆瘮杈，

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