Cui Luying, Ding Yi, Feng Yan, Chen Shuhuai, Xu Yingqing, Li Meng, Hu Manli, Qiu Zhengying, Ding Mingxing
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
Mol Neurobiol. 2017 Mar;54(2):1429-1439. doi: 10.1007/s12035-016-9759-8. Epub 2016 Feb 4.
Acupuncture tolerance is the gradual decrease in analgesic effect due to its prolonged application. However, its mechanism in terms of miRNA is still unknown. To explore the role of miRNAs in electroacupuncture (EA) tolerance of rats using deep sequencing, rats with more than a 50 % increase in tail flick latency (TFL) in response to EA were selected for this experiment. EA tolerance was induced by EA once daily for eight consecutive days. The hypothalami were harvested for deep sequencing. As a result, 49 differentially expressed miRNAs were identified and validated by real-time PCR. Of them, let-7b-5p, miR-148a-3p, miR-124-3p, miR-107-3p, and miR-370-3p were further confirmed to be related to EA tolerance by an intracerebroventricular injection of agomirs or antagomirs of these miRNAs. Potential targets of the 49 miRNAs were enriched in 9 pathways and 282 gene ontology (GO) terms. Five miRNAs were confirmed to participate in EA tolerance probably through the functional categories related to nerve impulse transmission, receptor signal pathways, and gene expression regulation, as well as pathways related to MAPK, neurotrophin, fatty acid metabolism, lysosome, and the degradation of valine, leucine, and isoleucine. Our findings reveal a characterized panel of the differentially expressed miRNAs in the hypothalamus in response to EA and thus provide a solid experimental framework for future analysis of the mechanisms underlying EA-induced tolerance.
针刺耐受是指由于针刺的长期应用导致镇痛效果逐渐降低。然而，其在微小RNA（miRNA）方面的机制仍不清楚。为了利用深度测序探索miRNA在大鼠电针（EA）耐受中的作用，本实验选取了对EA反应时甩尾潜伏期（TFL）增加超过50%的大鼠。通过连续八天每天一次EA诱导产生EA耐受。采集下丘脑进行深度测序。结果，鉴定出49个差异表达的miRNA，并通过实时聚合酶链反应进行了验证。其中，通过脑室内注射这些miRNA的激动剂或拮抗剂，进一步证实了let-7b-5p、miR-148a-3p、miR-124-3p、miR-107-3p和miR-370-3p与EA耐受有关。这49个miRNA的潜在靶标富集在9条通路和282个基因本体（GO）术语中。证实5个miRNA可能通过与神经冲动传递、受体信号通路、基因表达调控相关的功能类别，以及与丝裂原活化蛋白激酶（MAPK）、神经营养因子、脂肪酸代谢、溶酶体以及缬氨酸、亮氨酸和异亮氨酸降解相关的通路参与EA耐受。我们的研究结果揭示了下丘脑对EA反应时差异表达的miRNA特征谱，从而为未来分析EA诱导耐受的潜在机制提供了坚实的实验框架。
