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邱方,周杨,张严焱,金城,刘晓东,吴迎,吕媛媛,石丽君.miR-328 在运动改善高血压大鼠胸主动脉 L 型钙通道表达中的作用[J].中国实验动物学报,2020,28(6):749~758.
miR-328 在运动改善高血压大鼠胸主动脉 L 型钙通道表达中的作用
Effect of miR-328 in the exercise-induced improvement of the L-type calcium channel expression in thoracic aorta from hypertensive rats
投稿时间:2020-06-08  
DOI:10. 3969 / j.issn.1005-4847. 2020. 06. 003
中文关键词:  高血压  miR-328  有氧运动  LTCC  血管平滑肌细胞
英文关键词:hypertension  miR-328  aerobic exercise  LTCC  vascular smooth muscle cells
基金项目:
作者单位E-mail
邱方 北京体育大学运动生理教研室,北京 100084 2019112031@ bsu.edu.cn 
周杨 北京体育大学运动生理教研室,北京 100084  
张严焱 北京体育大学运动生理教研室,北京 100084  
金城 北京体育大学运动生理教研室,北京 100084  
刘晓东 北京体育大学运动生理教研室,北京 100084  
吴迎 1. 北京体育大学运动生理教研室,北京 100084
2. 北京体育大学运动与体质健康教育部重点实验室, 北京 100084 
 
吕媛媛 北京体育大学中国运动与健康研究院,北京 100084  
石丽君 1. 北京体育大学运动生理教研室,北京 100084
2. 北京体育大学运动与体质健康教育部重点实验室, 北京 100084 
shilj@ bsu.edu.cn 
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中文摘要:
       目的 探讨 microRNA-328(miR-328)在有氧运动改善高血压大鼠胸主动脉平滑肌细胞 L 型电压门控钙通道( voltage-gated L-type Ca2+ channel,LTCC) 表达的中促进作用。 方法 选取 12 周龄雄性正常血压大鼠 (Wistar Kyoto,WKY)与自发性高血压大鼠( spontaneously hypertensive rat,SHR) 随机分为正常血压对照组(WKY- C)、正常血压运动组(WKY-EX)、高血压对照组(SHR-C)、高血压运动组(SHR-EX),运动组进行 12 周中等强度跑 台训练。 12 周后采用股动静脉置管术检测大鼠心血管反应;取大鼠胸主动脉,采用 HE 染色检测胸主动脉形态, qPCR 检测 LTCC α1C和 β1 亚基 mRNA、miR-328 表达,Western Blot 检测 α1C和 β1 亚基蛋白表达。离体实验,培养 WKY 胸主动脉平滑肌细胞,脂质体转染 miR-328 mimic 和 miR-328 inhibitor 使之过表达或沉默,检测 LTCC α1C及 β1 亚基 mRNA 和蛋白表达变化。 结果 (1)有氧运动可显著降低 SHR 体重、血压和心率;(2)运动可显著降低 SHR 胸主动脉壁厚;(3)运动可显著降低 SHR 对去甲肾上腺素的升压反应和硝苯地平 Nifedipine(LTCC 阻断剂)的降压反应;(4)SHR-C 组的 LTCC α1C及 β1 亚基 mRNA、蛋白表达显著高于 WKY-C 组,而 SHR-EX 组的 α1C及 β1 亚基 mRNA、蛋白表达显著低于 SHR-C 组;(5)SHR-C 组中 miR-328 的表达显著低于 WKY-C 组,12 周有氧运动后 miR- 328 表达量显著增加;(6)离体实验中,与对照组相比,转染 miR-328 mimic 组 α1C和 β1 亚基 mRNA、蛋白表达显著下调,而转染 miR-328 inhibitor 组 α1C和 β1 亚基 mRNA、蛋白表达显著上调。 结论 规律有氧运动可有效降低 SHR 血压,调控 miR-328 靶向抑制 LTCC α1C和 β1 亚基的表达,可能是运动改善高血压胸主动脉 LTCC 重构的机制之一。
英文摘要:
       Objective The present study investigated the effect of microRNA-328 (miR-328) during the aerobic exercise-induced improvement of voltage-gated L-type Ca2+ channel ( LTCC) expression in thoracic aorta smooth muscle cells from spontaneously hypertensive rats ( SHRs). Methods Twelve-week-old male normotensive Wistar Kyoto rats (WKYs) and SHRs randomly were separated into a control group (SHR-C and WKY-C) and an exercise group (SHR-EX and WKY-EX). Rats in exercise were subjected to moderate-intensity treadmill training for 12 weeks. The cardiovascular responses of rats were then detected by femoral arterial and venous cannulation. The thoracic aorta was stained with hematoxylin and eosin ( HE) for histological assays. The mRNA expression levels of LTCC α1C and β1 subunits and the expression of miR-328 were detected by qPCR. The protein expression levels of LTCC α1C and β1 subunits were detected by Western Blot. For in vitro analysis, primary vascular smooth muscle cells (VSMCs) were isolated and cultured from the thoracic aorta of WKYs. Both miR-328 mimic and miR-328 inhibitor were transfected into cultured arterial myocytes to produce miR-328 overexpression or silencing. The mRNA and protein expression levels of LTCC α 1C and β1 subunits were detected. Results Aerobic exercise significantly reduced body weight, blood pressure, and heart rate in SHRs. Aerobic exercise significantly reduced the wall thickness of thoracic aorta in SHRs. Exercise significantly reduced the pressor response to norepinephrine and the depressor response to nifedipine ( LTCC blocker) in SHRs. The mRNA and protein expression levels of LTCC α1C and β1 subunits were significantly upregulated in the SHR-C group, compared with the WKY- C group, while the mRNA and protein expression levels of LTCC α1C and β1 subunits were downregulated in the SHR-EX group, compared with the SHR-C group. miR-328 expression was significantly lower in the SHR-C group than that in the WKY-C group; this expression significantly increased after aerobic exercise. In the transfection experiment, compared with the negative control ( NC) group, the mRNA and protein expression levels of α1C and β1 subunits were significantly downregulated in the miR-328 mimic transfected group; these expression was significantly upregulated in the miR-328 inhibitor transfected group. Conclusions Regular aerobic exercise may effectively reduce blood pressure in SHR and enable miR-328 to inhibit LTCC α1C and β1 subunit expression. This mechanism might contribute to exercise-mediated improvement of LTCC channel remodeling in hypertensive thoracic aorta.
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