[1]梁冰,董铁立.miR-19a对慢性压迫性损伤大鼠神经病理性疼痛的影响及其机制[J].第三军医大学学报,2018,40(07):590-595.
 LIANG Bing,DONG Tieli.Effect of miR-19a on neuropathic pain in chronic constriction injury rats and its mechanism[J].J Third Mil Med Univ,2018,40(07):590-595.
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miR-19a对慢性压迫性损伤大鼠神经病理性疼痛的影响及其机制(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第07期
页码:
590-595
栏目:
基础医学
出版日期:
2018-04-15

文章信息/Info

Title:
Effect of miR-19a on neuropathic pain in chronic constriction injury rats and its mechanism
作者:
梁冰董铁立
河南中医药大学第一附属医院麻醉科;郑州大学第二附属医院麻醉科
Author(s):
LIANG Bing DONG Tieli

Department of Anesthesiology, the First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan Province, 450099;Department of Anesthesiology, the Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, 450014, China

关键词:
miR-19a慢性压迫性损伤SOCS3机械性触诱发痛热痛觉过敏大鼠
Keywords:
miR-19a chronic constriction injury SOCS3 mechanical touch-evoked pain thermal hyperalgesia rats
分类号:
R362.21; R394.3; R441.1
文献标志码:
A
摘要:

目的    观察miR-19a对慢性压迫性损伤(chronic constriction injury,CCI)大鼠神经病理性疼痛的作用,并探讨其潜在机制。方法    75只雄性SD大鼠按随机数字表法分为3组:假手术组,行假手术;CCI组,坐骨神经结扎法构建CCI大鼠模型;CCI+ inhibitor组,CCI大鼠鞘内注射miR19a inhibitor。于术前当天(0 d)和术后3、7、14、21 d观察机械缩足反射阈值(mechanical withdrawal threshold,MWT)和热刺激反射潜伏期(thermal withdrawal latency,TWL)的变化。采用RT-qPCR检测腰椎脊髓中促炎因子IL-6、TNF-α、IL-1β mRNA及JAK2和STAT3表达的变化,Western blot检测SOCS3蛋白水平的表达,荧光素酶报告基因技术验证miR-19a的靶基因。结果     与假手术组比较,CCI组大鼠脊髓的miR-19a表达显著上升(P<0.05),MWT显著降低,TWL显著缩短(P<0.05)。与CCI组比较,CCI+inhibitor组MWT显著升高,TWL显著延长(P<0.05)。RT-qPCR检测结果表明,CCI可显著促进促炎因子IL-6、TNF-α、IL-1β mRNA的表达;而与CCI组比较,CCI+inhibitor组IL-6、TNF-α、IL-1β mRNA表达水平均显著降低(P<0.05)。另一方面,与假手术组比较,CCI组大鼠术后脊髓中JAK2和STAT3 mRNA水平明显增加;与CCI组比较,CCI+inhibitor组术后脊髓中JAK2和STAT3的表达显著降低(P<0.01)。荧光素酶报告基因检测结果表明JAK2/STAT3上游的负调控因子SOCS3是miR-19a的靶基因。Western blot检测结果表明CCI组大鼠脊髓中SOCS3水平较假手术组升高,且SOCS3水平在CCI术后第7天达到最高,而后呈下降趋势;与CCI组比较,CCI+ inhibitor组SOCS3表达明显上升(P<0.05)。结论    miR-19a inhibitor能缓解CCI大鼠机械性触诱发痛和热痛觉过敏,该作用与SOCS3介导的JAK2/STAT3通路有关。

Abstract:

Objective    To determine the effect of miR-19a on neuropathic pain in rats and investigate the potential mechanisms. Methods     A rat model of chronic constriction injury (CCI) was established by sciatic nerve ligation. A total of 75 male SD rats were randomly divided into 3 groups, that is, sham group, CCI group, CCI+inhibitor group (intrathecal injection of miR-19a inhibitor). The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) of each group were observed on the surgery day (regarded as 0 d) and 3, 7, 14 and 21 d post-operatively. Simultaneously, the mRNA expression levels of IL-6, TNF-α, IL-1β, JAK2 and STAT3 in the lumbar spinal cord were detected with RT-qPCR. Then, luciferase report gene assay was used to verify the target genes of miR-19a. Results     Compared to the sham group, miR-19a was highly expressed in the spinal cord tissue of CCI group (P<0.05), while MWT and TWL were significantly decreased (P<0.05). However, MWT and TWL were markedly increased in the CCI+inhibitor group when compared with the CCI group (P<0.05). What is more, RT-qPCR analysis showed that compared with the sham group, the pro-inflammatory factors, such as IL-6, TNF-α and IL-1β were highly increased in the CCI group, while they were decreased in the CCI+inhibitor group. Compared with the sham group, the mRNA expression levels of JAK2 and STAT3 were significantly enhanced in the spinal cord tissues of CCI rats, while inhibition of miR19a reversed these effects (P<0.05). Further luciferase assay identified that SOCS3, an upstream suppressor of JAK2/STAT3, was a target gene of miR-19a. Western blot assay showed the level of SOCS3 was highly expressed in the CCI group when compared with sham group, and it reached the highest at day 7 post-operatively. Compared with CCI group, the SOCS3 expression was markedly increased in CCI+inhibitor group (P<0.05). Conclusion     miR-19a inhibitor alleviates the mechanical withdrawal pain and thermal hyperalgesia, and this function is associated with SOCS3mediated JAK2/STAT3 pathway.

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更新日期/Last Update: 2018-04-10