MiR-124 靶向调控 CX3CR1 影响骨癌痛小鼠痛觉
| 作者: |
1胡焓,
1冯丹,
1李少军
1 武汉市第一医院疼痛科，武汉 430022 |
| 通讯: |
冯丹
Email: fengdanvipvd@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2019.04.005 |
| 基金: | 武汉市卫生和计划生育委员会科研项目 (WX17D04)。 |
摘要
目的：探讨microRNA-124(miR-124)与趋化因子受体CX3CR1的调控机制及其对骨癌痛( bone cancer pain，BCP)小鼠痛觉行为的影响。方法：将L i es肺癌细胞注入小鼠左侧胫骨的骨髓腔内，构建BCP模型；将健康雄性小鼠64只随机分为8组( n=8)：BCP组、Sh a n组、Sham+Veh组、BCP+Veh组、BCP+miR-NC组、BCP+miR-124 mimics组、BCP+CX3CR1in组、BCP+miR-124+CX3CR1组；分别检测各组小鼠的热缩足潜伏期(thermal w ithdrawal latenc y，TWL)和机械性缩足反射阈值(mechanical w ithdrawal threshold，MWT)；RT-qPCR和Western印迹法分别检测BCP小鼠中miR-124和CX3CR1的表达；双荧光素报告基因实验和Western印迹法验证miR-124和CX3CR1之间的靶向调控关系。结果：MiR-124在BCP小鼠中表达下调；过表达miR-124能提高其TWL和MWT值；CX3CR1是miR-124的靶基因；敲低CX3CR1可显著提高TWL和MWT值；过表达CX3CR1能够显著逆转miR-124上调对BCP小鼠痛觉行为的影响。结论：MiR-124负调控CX3CR1参与调控BCP小鼠的痛觉。
关键词：
骨癌痛；miR-124；CX3CR1；MWT；TWL
MiR-124 participates in pain in mice with bone cancer pain by targeting CX3CR1
CorrespondingAuthor: FENG Dan Email: fengdanvipvd@163.com
DOI: 10.3978/j.issn.2095-6959.2019.04.005
Foundation: This work was supported by the Health and Family Planning Commission of Wuhan Municipality, China (WX17D04).
Abstract
Objective: To investigate the regulatory mechanism of microRNA-124 (miR-124) and CX3C chemokine receptor 1 (CX3CR1) and their effects on pain behaviors in mice with bone cancer pain (BCP). Methods: The Lies lung cancer cells were injected into the bone marrow cavity of the left tibia of the mouse to establish a BCP model. Sixty-four healthy male mice were divided into eight groups (n=8): BCP group, Sham group, Sham+Veh group, BCP+Veh group, BCP+miR-NC group, BCP+miR-124 group, BCP+CX3CR1in group, BCP+miR-124+CX3CR1 group. RT-qPCR and Western blot were performed to measure the levels of miR-124 and CX3CR1 in BCP mice. Thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) assays were conducted to determine heat pain threshold and mechanical withdrawal threshold, respectively. The relationship between miR-124 and CX3CR1 was confirmed by luciferase reporter and western blot assays. Results: The level of miR-124 was downregulated in BCP mice. Moreover, overexpression of miR-124 increased the values of TWL and MWT. CX3CR1 is a direct target gene of miR-124. CX3CR1 knockdown significantly elevated values of TWL and MWT. Interestingly, upregulation of CX3CR1 undermined the effects of miR-124 overexpression on pain in BCP mice. Conclusion: miR-124 was involved in pain of BCP mice via inhibiting CX3CR1.
Keywords：
bone cancer pain; miR-124; CX3CR1; mechanical withdrawal threshold; thermal withdrawal latency