中国中药杂志

2019, v.44(08) 1642-1647

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乌头碱抑制血管紧张素Ⅱ诱导的心肌细胞肥大
Aconitine ameliorates cardiomyocyte hypertrophy induced by angiotensin Ⅱ

王宁宁;王佳;谭洪玲;王宇光;高月;马增春;
WANG Ning-ning;WANG Jia;TAN Hong-ling;WANG Yu-guang;GAO Yue;MA Zeng-chun;Guangdong Pharmaceutical University;Institute of Radiation Medicine,Academy of Military Medical Sciences;

摘要(Abstract):

研究乌头碱(aconitine,AC)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导H9c2细胞肥大的抑制作用,并探讨其作用机制。通过AngⅡ诱导H9c2细胞肥大建立模型,并分别与不同浓度的乌头碱共同处理,Western blot法检测atrial natriuretic peptide(ANP)、brain natriuretic peptide(BNP)、β-myosin heavy chain(β-MHC)、α-smooth muscle actin(α-SMA)在蛋白水平的表达;实时荧光定量PCR技术(qRT-PCR)检测肥大基因ANP,BNP,β-MHC mRNA的表达。采用激光扫描共聚焦显微镜检测肌原纤维中的重要组成成分F-actin标记的荧光强度。乌头碱可明显逆转AngⅡ所诱导的H9c2细胞总蛋白含量的升高; qRTPCR检测结果显示乌头碱可明显抑制AngⅡ所诱导的ANP,BNP,β-MHC mRNA的上调; Western blot法检测提示乌头碱可明显抑制AngⅡ所诱导的ANP,BNP,β-MHC蛋白的上调;荧光标记检测F-actin的表达水平,乌头碱可以抑制AngⅡ所诱导的F-actin的表达。乌头碱明显下调AngⅡ所诱导的心肌细胞肥大的多项指标,揭示乌头碱可能通过抑制肥大因子的上调来缓解AngⅡ所引起的心肌肥大,这可能是乌头碱发挥治疗作用的机制之一。
This paper was aimed to investigate the inhibitory effect of aconitine(AC) on angiotensin Ⅱ(Ang Ⅱ)-induced H9 c2 cell hypertrophy and explore its mechanism of action. The model of hypertrophy was induced by Ang Ⅱ(1×10-6 mol·L-1),and cardiomyocytes were incubated with different concentrations of AC. Western blot was used to quantify the protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and α-smooth muscle actin(α-SMA). Real-time quantitative PCR(qRT-PCR) was used to quantify the mRNA expression levels of cardiac hypertrophic markers ANP,BNP and β-MHC. In addition,the fluorescence intensity of the F-actin marker,an important component of myofibrils,was detected by using laser confocal microscope. AC could significantly reverse the increase of total protein content in H9 c2 cells induced by Ang Ⅱ; qRT-PCR results showed that AC could significantly inhibit the ANP,BNP and β-MHC mRNA up-regulation induced by AngⅡ. Western blot results showed that AC could significantly inhibit the ANP,BNP and β-MHC protein up-regulation induced by AngⅡ. In addition,F-actin expression induced by Ang Ⅱ could be inhibited by AC,and multiple indicators of cardiomyocyte hypertrophy induced by Ang Ⅱ could be down-regulated,indicating that AC may inhibit cardiac hypertrophy by inhibiting the expression of hypertrophic factors,providing new clues for exploring the cardiovascular protection of AC.

关键词(KeyWords): 乌头碱;血管紧张素Ⅱ;心肌肥大;H9c2细胞
aconitine;angiotensin Ⅱ;cardiac hypertrophy;H9c2 cells

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(81673633);国家自然科学基金重点项目(81630102)

作者(Authors): 王宁宁;王佳;谭洪玲;王宇光;高月;马增春;
WANG Ning-ning;WANG Jia;TAN Hong-ling;WANG Yu-guang;GAO Yue;MA Zeng-chun;Guangdong Pharmaceutical University;Institute of Radiation Medicine,Academy of Military Medical Sciences;

DOI: 10.19540/j.cnki.cjcmm.20190117.002

参考文献(References):

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