慢病毒介导的DREAM沉默治疗在坐骨神经慢性压迫模型大鼠疼痛中的作用及其对GALR1的表达调控

发布时间：2019-01-02 08:04

【摘要】：目的甘丙肽受体1(galanin receptor 1,GALR1)信号通路在痛觉调节中发挥重要作用,该研究利用坐骨神经慢性压迫(chronic constriction injury of the sciatic nerve,CCI)大鼠模型,探索慢病毒介导的DREAM沉默治疗在CCI模型大鼠疼痛中的作用及其对GALR1的表达调控。方法选择健康雄性SD大鼠24只,随机分为4组(n=6):即RNA干扰组、空白载体组、单纯CCI组和正常对照组。鞘内置管前后分别测定基础痛阈,RNA干扰组和空白载体组于CCI后经微导管给予pKCSHR-Puro/GFP-DREAM慢病毒和空白载体,并测定腰段脊髓内下游调控元件拮抗分子(downstream regulatory element antagonist,DREAM)和GALR1的蛋白表达。结果大鼠痛阈的变化:手术同侧热痛阈和机械痛阈测定结果表明,CCI处理后,RNA干扰组、空白载体组及单纯CCI组在各个时间点热痛阈和机械痛阈较正常对照组均显著降低(P0.01);RNA干扰组鞘内注射后较注射前痛阈显著升高,治疗后相同时间点RNA干扰组较空白载体组及单纯CCI组痛阈亦显著升高。Western blotting结果表明,与其他实验组对比,RNA干扰组的DREAM蛋白表达水平显著下调,而GALR1蛋白表达水平较空白载体组、单纯CCI组亦有显著下调。结论 DREAM可以调控GALR1的表达,提示甘丙肽受体1信号通路参与DREAM基因调节大鼠神经病理性疼痛。
[Abstract]:Objective to study the effect of glycine receptor 1 (galanin receptor 1 (GALR1) signal pathway on the regulation of pain in rat model of chronic compression of sciatic nerve (chronic constriction injury of the sciatic nerve,CCI). To explore the role of lentivirus-mediated DREAM silencing in pain in CCI model rats and the regulation of GALR1 expression. Methods 24 healthy male SD rats were randomly divided into 4 groups: RNA interference group, blank vector group, simple CCI group and normal control group. The basal pain threshold was measured before and after intrathecal catheterization. The RNA interference group and blank carrier group were treated with pKCSHR-Puro/GFP-DREAM lentivirus and blank vector via microcatheter after CCI, and the antagonist molecule (downstream regulatory element antagonist, of the downstream regulatory element in the lumbar spinal cord was measured. DREAM) and GALR1 protein expression. Results the changes of pain threshold in rats: the results of ipsilateral thermal pain threshold and mechanical pain threshold measurement showed that after CCI treatment, RNA interference group, The thermal pain threshold and mechanical pain threshold of blank carrier group and simple CCI group were significantly lower than those of normal control group at each time point (P0.01). The pain threshold of RNA interference group was significantly higher than that of pre-injection group, and the pain threshold of RNA interference group was significantly higher than that of blank carrier group and CCI group at the same time after treatment. The results showed that the pain threshold of RNA interference group was significantly higher than that of other experimental groups. The expression level of DREAM protein in RNA interference group was significantly down-regulated, while GALR1 protein expression level was significantly lower than that in blank vector group and CCI alone group. Conclusion DREAM can regulate the expression of GALR1, suggesting that glycidyl receptor 1 signaling pathway is involved in the regulation of neuropathic pain by DREAM gene in rats.
【作者单位】：湖南省人民医院麻醉科;中南大学湘雅医院麻醉科;
【基金】：国家自然科学基金(No:30872427) 湖南省自然科学基金(No:05JJ40136) 湖南省科技计划资助项目(No:2012SK3241)
【分类号】：R-332

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