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张诗渊.miR-181b 调控 Mex3B 蛋白对动脉粥样硬化斑块形成及分子机制研究[J].中国比较医学杂志,2020,30(8):76~85.
miR-181b 调控 Mex3B 蛋白对动脉粥样硬化斑块形成及分子机制研究
miR-181b regulates atherosclerotic plaque formation by targeting Mex3B expression
投稿时间:2020-01-02  
DOI:10. 3969 / j.issn.1671-7856. 2020. 08. 012
中文关键词:  miR-181b  动脉粥样硬化  肌肉过量蛋白-3B  大鼠
英文关键词:miR-181b  atherosclerosis  Mex3B  rat
基金项目:
作者单位E-mail
张诗渊 南昌大学第二附属医院超声科,南昌 330006 p3gnmt@ 163.com 
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中文摘要:
      目的 探讨 miR-181b 靶向调控肌肉过量蛋白-3B(muscle excess protein-3,Mex3B)的表达在动脉粥 样硬化(atherosclerosis,AS)斑块的形成及其分子作用机制。 方法 将雄性 SD 大鼠随机分成正常组、模型组、对照组和实验组。对照组和实验组分别尾静脉注射 miR-181b-inhibitor-NC、miR-181b-inhibitor 的混合液,正常组、模型组注射等剂量的生理盐水;注射完毕 24 h 后将维生素 D3 以 600000 U/ kg 的剂量一次性腹腔注射进模型组、对照组、 实验组大鼠体内后,每日以 100 g 高脂饲料喂养构建 AS 模型大鼠,正常组大鼠始终以等量基础饲料喂养。 喂养 60 d后,对大鼠进行颈动脉超声检查,测量管腔动脉后壁内膜-中层厚度( intima-media thickness of the posterior wall of the artery,IMT)和斑块面积( square, S) 参数。 RT-PCR 检测各组 miR-181b 和 Mex3B 蛋白的 mRNA 的表达; Western blot 检测各组 Mex3B 蛋白的表达水平;荧光素酶报告基因分析 miR-181b 和 Mex3B 的调控关系。 结果 与 正常组相比,模型组大鼠颈动脉内膜增生明显,颈动脉 IMT、斑块 S、miR-181b 和 Mex3B 蛋白的 mRNA 和蛋白的表达明显增大,与模型组相比,实验组大鼠大鼠颈动脉内膜增生明显减小,颈动脉 IMT、斑块 S、miR-181b 和 Mex3B 蛋 白的 mRNA 和蛋白的表达明显减小,差异均具有统计意义(P<0. 05)。 荧光素酶报告基因分析证明 miR-181b 与 Mex3B 靶向结合。 结论 miR-181b 通过靶向调控 Mex3B 的表达参与动脉粥样硬化中的炎性应激反应,抑制 miR- 181b 的表达能明显抑制炎症反应,抑制 AS 中斑块的形成。
英文摘要:
      Objective We investigated the role of miR-181b in regulating atherosclerotic plaque formation in atherosclerosis (AS) by targeting Mex3B expression. Methods Male Sprague-Dawley rats were randomly divided into a normal group, model group, control group, and experimental group. A mixture of miR-181b-inhibitor-NC and miR-181b- inhibitor was injected into the control group and the experimental group, while rats in the normal group and model group received the same amount of saline. At 24 hours after the injection, a 600, 000 U/ kg dose of vitamin D3 was intraperitoneally injected into the rats in the model group, control group, and experimental group, which then received a daily 100 g high-fat diet. The rats in the normal group always received the same amount of a basic diet. After 60 days of feeding, the rats underwent carotid ultrasound examination to measure the intima-media thickness of the posterior wall of the artery (IMT) and the plaque area (square, S). RT-PCR was used to detect miR-181b expression and the mRNA of Mex3B protein, and a Western blot was used to detect the expression of Mex3B protein. A luciferase reporter gene analysis was used to examine the regulatory relationship between miR-181b and Mex3B. Results Compared with the normal group, carotid intima hyperplasia was evident in the model group, and we observed significant increases in carotid IMT, plaque S, miR-181b and Mex3B mRNA expression, and Mex3B protein expression (P< 0. 05). Compared with the model group, carotid intima hyperplasia was evident in the experimental group, and we observed significant decreases in carotid IMT, plaque S, miR-181b and Mex3B mRNA expression, and Mex3B protein expression (P< 0. 05). Luciferase reporter gene analysis demonstrated that miR-181b targets Mex3B. Conclusions miR-181b is involved in the inflammatory stress response in atherosclerosis by targeting Mex3B expression. Inhibition of miR-181b expression may significantly inhibit inflammatory responses and reduce plaque formation in AS.
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