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邢春蕊,陈应奇,孙 曌,刘英莲.针刺通过调节 GABA-Gln 代谢对失眠大鼠症状的改善及对自发活动的影响[J].中国比较医学杂志,2021,31(5):47~52.
针刺通过调节 GABA-Gln 代谢对失眠大鼠症状的改善及对自发活动的影响
Improvement of symptoms and spontaneous activity of insomniac rats by regulating GABA-Gln metabolism
投稿时间:2020-08-06  
DOI:10. 3969 / j.issn.1671-7856. 2021. 05. 008
中文关键词:  失眠  针刺  γ-氨基丁酸  谷氨酸
英文关键词:insomnia  acupuncture  γ-aminobutyric acid  glutamic acid
基金项目:
作者单位E-mail
邢春蕊 海口市人民医院 健康管理体检科, 海口 570802 chon4521g@ 163.com 
陈应奇 海南医学院中医学院,海口 570802 chon4521g@ 163.com 
孙 曌 海南医学院中医学院,海口 570802  
刘英莲 海南医学院中医学院,海口 570802  
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中文摘要:
       目的 研究针刺对失眠大鼠症状的改善及对自发活动的影响,并探讨其可能作用机制。 方法 腹腔注射对氯苯丙氨酸(PCPA)建立失眠大鼠模型,随机分为失眠组、针刺组和西药组,各 10 只,另设对照组 10 只。 针刺组和西药组分别采用针刺及灌胃地西泮片进行干预,失眠组和对照组固定后灌胃生理盐水,干预 7 d。 旷场实验检测自发活动;戊巴比妥钠协同睡眠实验检测睡眠潜伏期和睡眠时间;RT-qPCR 检测视交叉上核生物钟基因 clockperiod2(per2)水平;高效液相检测下丘脑 γ-氨基丁酸(GABA)、谷氨酸(Glu)含量;Western blot 检测下丘脑 GAD、GS 蛋白表达水平。 结果 模型组大鼠出现失眠典型症状。 与模型组比较,针刺组和西药组水平运动和垂直 运动得分升高,睡眠潜伏期缩短、睡眠时间延长,视交叉上核 clockper2 mRNA 相对表达量升高,下丘脑 Glu 水平降低、GABA 水平升高、Glu / GABA 降低,GAD 蛋白相对表达量升高、GS 蛋白相对表达量降低(P<0. 001,P<0. 01,P< 0. 05);针刺组与西药组上述指标比较,差异无统计学意义(P>0. 05)。 结论 针刺对失眠大鼠症状及自发活动具有一定改善作用,可能通过调节 GABA-Gln 发挥作用。
英文摘要:
       Objective To investigate the role of microRNA (miR)-20a-5p regulating vascular endothelial growth factor (VEGF) pathway in mice with oxygen-induced retinopathy (OIR). Methods The experiment was divided into normal group, model group, hyperoxia control group, miR-20a-5p high expression group, with 24 mice in each group. In addition to the normal group, the rest of the mice were placed in the oxygen tank with ( 75. 00 ± 2. 00)% oxygen concentration from the 7th day after birth to establish the OIR mice model, and after 5 days of continuous hyperoxia environment, they were fed in normoxia, at 1 day before the end of hyperoxia environment, 1 μL phosphate buffered saline (PBS) was injected into the vitreous cavity of the hyperoxia control group, 1 μL miR-20a-5p agomir (1 μmol / L) was injected into the vitreous cavity of the high expression group, and the model group did nothing. After the end of hyperoxia, the normal group was kept in the air for normal feeding. The experiment was carried out after normal air feeding for more 5 days. The expressions of miR-20a-5p, VEGF, vascular endothelial growth factor receptor (VEGFR)-1 and VEGFR-2 were detected by real-time fluorescence quantitative PCR ( qRT-PCR); retinal vascular morphology was observed by retinal patch; hematoxylin eosin (HE) staining was used to count the endothelial cells of retinal neovascularization; and VEGF positive cells were detected by immunohistochemistry. Results In the model group and hyperoxia control group, on the 17th day of life, the radial large blood vessels from the optic disc extended roundly and irregularly, there were a lot of new blood vessels, the structure and distribution of neovascularization were disordered and the capillary network was occluded; and compared with the model group and the hyperoxia control group, the miR-20a-5p high expression group had no obvious radial vascular circuitous, less irregular vascular expansion and less neovascularization. Compared with those in the normal group, the level of miR-20a-5p in retina in model group and hyperoxia control group was lower (P< 0. 05), and the number of nuclei in retinal vascular endothelium, VEGF protein positive area percentage, expressions of VEGF, VEGFR-1 and VEGFR-2 mRNAs in retina were higher (P< 0. 05); meanwhile, compared with those in the model group and the hyperoxia control group, the level of miR-20a-5p in retina in miR-20a-5p high expression group was lower (P< 0. 05), and the number of nuclei in retinal vascular endothelium, VEGF protein positive area percentage, expressions of VEGF, VEGFR-1 and VEGFR-2 mRNAs in retina were higher ( P< 0. 05). Conclusions Increasing miR-20a-5p can inhibit VEGF pathway and decrease retinal neovascularization in OIR mice, so as to protect the retina of OIR mice.
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