p15与c-Myc在TGF-β1诱导的大鼠心肌细胞肥大中的表达及与TGF/Smad信号通路的关系

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p15与c-Myc在TGF-β1诱导的大鼠心肌细胞肥大中的表达及与TGF/Smad信号通路的关系
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作者单位:(1.广州医科大学附属第一医院老年科 广州心血管病研究所,广东省广州市 510120;2.广州胸科医院神经内科,广东省广州市 510095;3.广州医科大学附属第一医院老年科,广东省广州市 510120;4.广州医科大学附属第二医院心内科广州心血管病研究所,广东省广州市510260;5.南方医科大学附属珠江医院心血管病实验室,广东省广州市 510515)
作者简介:石永英,博士,副主任医师,研究方向为高血压左心室肥厚的预防与治疗,E-mail为shiyongying121@163.com。
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基金项目:广东省医学科学技术研究基金项目(A2016376)

Study on p15 and c-Myc with TGF/Smad signal pathway of cardiomyocyte hypertrophy induced by TGF-β1 in rat

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1.Department of Geriatrics, the First Affiliated Hospital of Guangzhou Medical University & Guangzhou Institute of Cardiovascular Disease, Guangzhou, Guangdong 510120, China;2.Department of Neurology, Guangzhou Chest Hospital, Guangzhou, Guangdong 510095, China;3.Department of Geriatrics, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, China;4.Department of Cardiology, the Second Affiliated Hospital of Guangzhou Medical University & Institute of Cardiovascular Disease, Guangzhou, Guangdong 510260, China;5.Institute of Cardiovascular Disease, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China)

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目的探讨转化生长因子β1(TGF-β1)诱导的大鼠心肌细胞肥大中Smad信号通路及其目的蛋白p15的表达。方法建立TGF-β1诱导的体外大鼠心肌细胞肥大模型。PI染色标记细胞双链RNA法检测心肌细胞RNA表达量,间接检测心肌细胞肥大。实时荧光定量PCR检测心肌细胞肥大相关基因β肌球蛋白重链(β-MHC)的表达。Smad2 siRNA干扰Smad信号通路表达并检测其相关蛋白表达。 Western blot检测心肌细胞p-Smad2、Smad2、Smad2/3、p15及c-Myc蛋白表达。结果 TGF-β1可诱导体外培养的心肌细胞肥大基因的表达,β-MHC的表达明显高于对照组(P＜0.01)。PI染色标记细胞双链RNA法检测发现,TGF-β1组RNA含量明显增高(P＜0.01),与对照组相比,TGF-β1可明显上调p-Smad2、Smad2、Smad2/3及Smad通路目的蛋白p15、c-Myc的表达(P＜0.01)。给予Smad2 siRNA特异性干扰后,p15表达较TGF-β1组减少(P＜0.01)。结论 TGF-β1可能通过Smad蛋白通路诱导心肌细胞肥大形成,此过程中相关目的蛋白p15 表达增加。

Abstract:

Aim To investigate pathway mechanism about transforming growth factor-β1 (TGF-β1) activated cardiomyocyte hypertrophy. Methods Fresh isolated neonatal rat cardiomyocytes induced by TGF-β1 were used to examine formation of cell pathway. p-Smad2, Smad2, Smad2/3, p15 and c-Myc proteins were measured by Western blot. Cardiomyocyte hypertrophy was measured by PI statining and RNA expressions by real-time PCR. Smad signal pathway expression and it's related protein expression were induced by TGF-β1 and siRNA interference. Results TGF-β1 could induce the expression of hypertrophic gene in cultured cardiomyocytes in vitro, and the expression of β-MHC was significantly higher than that of the control group (P＜0.01). The content of RNA in TGF-β1 group was significantly increased by PI staining (P ＜0.01). Compared with the control group, TGF-β1 significantly increased the expression of p15 and c-Myc in p-Smad2, Smad2, Smad2/3 and Smad pathway (P＜0.01). The expression of p15 of Smad2 siRNA group was less than that of TGF-β1 group (P＜0.01). Conclusion TGF-β1 may induce cardiomyocyte hypertrophy through the Smad protein pathway, and in this process the expression of p15 is increased.

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石永英,卢俊江,陈广原,刘世明,陈敏生. p15与c-Myc在TGF-β1诱导的大鼠心肌细胞肥大中的表达及与TGF/Smad信号通路的关系[J].中国动脉硬化杂志,2018,26(3):245~252.

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收稿日期:2017-09-27
最后修改日期:2017-11-02
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在线发布日期: 2018-04-03

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