MiR-139-3p通过靶向下调TRPV1缓解大鼠神经病理性疼痛

发布时间：2019-03-25 21:54

【摘要】：目的:研究miR-139-3p在大鼠神经病理性疼痛发生发展中的作用及其作用机制。方法:将大鼠随机分为假手术组(sham)、坐骨神经慢性压迫损伤模型组(chronic constriction injury group,CCI)、2 mg/kg miR-139-3p模拟物(mimic)鞘内注射组、4 mg/kg miR-139-3p mimic鞘内注射组。术后的第14 d,Real-time PCR法检测大鼠背根神经节(DRG)中miR-139-3p及TRPV1的表达。于注射前及注射后的第1、3、7、14 d,利用von Frey检测大鼠机械性痛阈及热辐射法检测大鼠热痛阈值。Western Blot法检测DRG中TRPV1的表达;ELISA法检测脊髓L4-L5组织中TNF-α及IL-1β的含量;双荧光素酶报告基因法检测miR-139-3p与TRPV1的靶向关系。结果:(1)与sham组相比较,miR-139-3p在CCI大鼠DRG中的表达显著下降,而TRPV1的表达显著上升(P0.05)。(2)与sham组相比较,CCI组中大鼠机械刺激缩足反射阈值及热刺激缩足反射潜伏期显著下降(P0.05);而与CCI组相比较,鞘内注射miR-139-3p mimic可显著促进大鼠机械刺激缩足反射阈值及热刺激缩足反射潜伏期的上升,抑制TRPV1的表达(P0.05)。(3)与sham组相比较,CCI组大鼠TNF-α及IL-1β含量显著上升(P0.05);而与CCI组相比较,鞘内注射miR-139-3p mimic可显著抑制TNF-α及IL-1β的产生(P0.05)。(4)miR-139-3p mimic转染可显著降低荧光素酶报告基因的荧光强度。结论:miR-139-3p通过靶向下调TRPV1的表达缓解大鼠神经病理性疼痛。
[Abstract]:Aim: to study the role and mechanism of miR-139-3p in the occurrence and development of neuropathic pain in rats. Methods: rats were randomly divided into sham operation group (chronic constriction injury group,CCI), (sham), chronic sciatic nerve compression injury model group (chronic constriction injury group,CCI), 2 mg/kg miR-139-3p analogue (mimic) intrathecal injection group, and 4 mg/kg miR-139-3p mimic intrathecal injection group. On the 14th day after operation, the expression of miR-139-3p and TRPV1 in (DRG) of rat dorsal root ganglion was detected by real time PCR. Before injection and 1,3,7,14 days after injection, the mechanical pain threshold and thermal pain threshold were detected by von Frey, TRPV1 expression in DRG, TNF- 伪 and IL-1 尾 in spinal cord L4-L5 tissue were detected by Western Blot and ELISA method, respectively, and the levels of TNF- 伪 and IL-1 尾 in spinal cord L4-L5 were measured by Western Blot and thermal radiation respectively at the 1st, 3rd, 7th and 14th day after injection. Double luciferase reporter gene method was used to detect the target relationship between miR-139-3p and TRPV1. Results: (1) compared with sham group, the expression of miR-139-3p in DRG of CCI rats decreased significantly, while the expression of TRPV1 increased significantly (P0.05). (2) compared with sham group. In CCI group, the threshold of foot contraction reflex induced by mechanical stimulation and the latency of foot contraction reflex induced by thermal stimulation were significantly decreased (P0.05). Compared with CCI group, intrathecal injection of miR-139-3p mimic significantly increased the threshold of foot contraction reflex induced by mechanical stimulation and the latency of foot contraction reflex induced by thermal stimulation, and inhibited the expression of TRPV1 (P0.05). (3) compared with sham group. The contents of TNF- 伪 and IL-1 尾 in CCI group were significantly higher than those in control group (P0.05). Compared with CCI group, intrathecal injection of miR-139-3p mimic significantly inhibited the production of TNF- 伪 and IL-1 尾 (P0.05). (4). MiR-139-3p mimic transfection significantly decreased the fluorescence intensity of luciferase reporter gene. Conclusion: miR-139-3p can alleviate neuropathic pain in rats by targeting down-regulating the expression of TRPV1.
【作者单位】：广西中医药大学附属瑞康医院神经内科;广西中医药大学附属瑞康医院麻醉科;
【分类号】：R741

【相似文献】