Mechanism of Mongolian medical warm acupuncture in treating insomnia by regulating miR-101a in rats with insomnia.
Mechanism of Mongolian medical warm acupuncture in treating insomnia by regulating miR-101a in rats with insomnia.
复制标题
DOI：
10.3892/etm.2017.4452
复制
发表时间：
2017-07
影响因子：
2.7
通讯作者：
Yuan H
中科院分区：
文献类型：
--
作者：
Bo A;Si L;Wang Y;Bao L;Yuan H
在基因水平上检测温针治疗前后microRNAs（miRNAs或miRs）及靶基因的表达，并研究失眠相关的细胞因子和神经递质。采用雄性Sprague-Dawley大鼠建立PCPA失眠大鼠模型，随机分为正常组、模型组、温针组和药物组。蒙药温针组针刺定会穴、合益穴、心穴。采用蒙药温针治疗失眠大鼠，观察治疗前后失眠大鼠脑组织microRNA的差异表达，建立miR-101 a模拟物和抑制剂。qPCR检测miR-101 a的表达水平。Western blotting检测PAX 8的表达水平。蒙药温针组与正常组比较，差异表达的miRNAs有141个。miR-101 a模拟物转染失眠大鼠后72 h海马细胞中miR-101 a的表达水平显著升高（P<0. 05）。转染miR-101 a抑制剂的神经元细胞活性在72 h时显著升高（P<0.05）。western blotting结果显示，与scramble组相比，miR-101 a mimics组失眠模型大鼠神经细胞PAX 8蛋白表达在72 h时受到抑制，表达下调（P<0. 01）。与空白对照组相比，下丘脑、海马和前额叶皮质中IL-1、IL-2、IL-6和肿瘤坏死因子-α水平均显著降低（P<0.05）。温针组和西药组治疗后去甲肾上腺素、多巴胺、谷氨酸水平均显著降低（P<0.05）。总之，本研究表明，在温针灸治疗的大鼠中，miR-101 a的上调与PAX 8调节直接相关。
MicroRNAs (miRNAs or miRs) and the target genes before and after warm acupuncture at the genetic level were assessed, and the cytokines and neurotransmitters related to insomnia were studied. Male Sprague-Dawley rats were used to create PCPA insomnia rat models and randomly divided into the normal, model, warm acupuncture, and drug groups. The Dinghui Acupoint, Heyi Acupoint, and Xin Acupoint were inserted in the Mongolian medicine warm acupuncture group. The differential expression profile of microRNA in the brain tissue of the insomnia rats was determined before and after Mongolian medicine warm acupuncture for establishment of miR-101a mimics and inhibitor. qPCR was used to detect the expression level of miR-101a. Western blotting was used to detect the expression level of PAX8. The rats receiving Mongolian medicine warm acupuncture had 141 miRNAs with differential expression compared with the normal rats. The expression level of miR-101a in the cells of the hippocampus of the insomnia rats transfected with miR-101a mimics increased significantly at 72 h (P<0.05). The activity of the neuronal cells transfected with miR-101a inhibitor increased significantly at 72 h (P<0.05). The western blotting result indicated that the expression of the PAX8 protein in the neuronal cells of the insomnia model rats was inhibited and downregulated significantly at 72 h after addition of miR-101a mimics compared with that in the scramble added group (P<0.01). The levels of the interleukins IL-1, IL-2, and IL-6 and the tumor necrosis factor-α in the hypothalamus, hippocampus, and prefrontal cortex decreased significantly compared with those in the blank control group (P<0.05). The levels of noradrenaline, dopamine, and glutamic decreased significantly following warm acupuncture or western medicine treatment (P<0.05). In conclusion, this study demonstrates that the upregulation of miR-101a in the rats treated with warm acupuncture is directly associated with PAX8 regulation.