Liu Xing, Geng Jiefeng, Bu Huilian, Ma Junqi, Ai Yanqiu
Department of Anesthesiology, Pain and Perioperative Medicine, The First Affiliated Hospital of Zhengzhou University No. 1, East Jianshe Rd Zhengzhou 450052 China
Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University Zhengzhou China.
RSC Adv. 2019 Dec 11;9(70):40895-40902. doi: 10.1039/c9ra06264e. eCollection 2019 Dec 9.
: Morphine is a commonly used analgesic drug. However, long-term use of morphine will cause tolerance which limits its clinical application in pain treatment. MicroRNAs (miRNAs) have been reported to be involved in the morphine tolerance, but the underlying mechanism is still poorly understood. : Tail flick test was used to measure the maximum possible effect (MPE). Quantitative real-time PCR was employed to detect miR-26b, BDNF, and Wnt5a expression in rat dorsal root ganglia (DRG). Luciferase report assay was introduced to verify the binding relationship between miR-26b and Wnt5a. BDNF, Wnt5a and β-catenin protein level were tested by western blotting. : MiR-26b was down-regulated during the development of morphine tolerance while BDNF was upregulated. Overexpression of miR-26b or BDNF inhibition alleviated morphine tolerance. Wnt5a was directly targeted and inhibited by miR-26b binding to the 3'-UTR of Wnt5a. The Wnt/β-catenin pathway was active in morphine tolerant rats. Moreover, overexpression of Wnt5a could partially enhance miR-26 mimic-mediated morphine tolerance, while a Wnt5a inhibitor could attenuate the tolerance. : The present study demonstrated that miR-26b overexpression alleviated morphine tolerance by inhibiting BDNF the Wnt/β-catenin pathway in rats, highlighting a promising target for the treatment of morphine tolerance.
吗啡是一种常用的镇痛药物。然而,长期使用吗啡会导致耐受性,这限制了其在疼痛治疗中的临床应用。据报道,微小RNA(miRNA)参与了吗啡耐受性的形成,但其潜在机制仍知之甚少。
采用甩尾试验测量最大可能效应(MPE)。运用定量实时聚合酶链反应检测大鼠背根神经节(DRG)中miR-26b、脑源性神经营养因子(BDNF)和Wnt5a的表达。引入荧光素酶报告基因检测以验证miR-26b与Wnt5a之间的结合关系。通过蛋白质免疫印迹法检测BDNF、Wnt5a和β-连环蛋白的蛋白水平。
在吗啡耐受性形成过程中,miR-26b表达下调,而BDNF表达上调。miR-26b过表达或BDNF抑制可减轻吗啡耐受性。miR-26b通过与Wnt5a的3'-非翻译区(3'-UTR)结合直接靶向并抑制Wnt5a。Wnt/β-连环蛋白信号通路在吗啡耐受大鼠中处于激活状态。此外,Wnt5a过表达可部分增强miR-26模拟物介导的吗啡耐受性,而Wnt5a抑制剂可减轻耐受性。
本研究表明,miR-26b过表达通过抑制BDNF及Wnt/β-连环蛋白信号通路减轻大鼠吗啡耐受性,为吗啡耐受性的治疗提供了一个有前景的靶点。