Qiliqiangxin Attenuates Phenylephrine-Induced Cardiac Hypertrophy through Downregulation of MiR-199a-5p
Qiliqiangxin Attenuates Phenylephrine-Induced Cardiac Hypertrophy through Downregulation of MiR-199a-5p
复制标题
七力强心通过下调 MiR-199a-5p 减轻去氧肾上腺素诱导的心脏肥大
DOI：
10.1159/000443113
复制
发表时间：
2016-01-01
影响因子：
--
通讯作者：
Li, Xinli
中科院分区：
文献类型：
--
作者：
Zhang, Haifeng;Li, Shanshan;Li, Xinli
背景/目的：芪苈强心（QL）是一种传统中药，长期用于治疗慢性心力衰竭。以往的研究表明，QL可以预防高血压或缺血应激引起的心脏重塑和肥大。然而，关于QL是否可以在体外调节心脏肥大，以及（如果是这样的话）是否通过调节特定的肥大相关microRNA，我们知之甚少。研究方法：分离、培养原代乳鼠心室肌细胞，并在有或无QL（0.5 μg/ml，48 h）预处理的情况下，用苯肾上腺素（PE，50 μmol/L，48 h）诱导体外肥大。α-辅肌动蛋白免疫荧光染色测定细胞表面积。qRT-PCR检测心肌肥厚标志物心钠素（ANP）、脑钠素（BNP）和β-肌球蛋白重链（MYH 7）的mRNA水平。心肌细胞的蛋白质合成通过蛋白质/DNA比率来确定。在PE处理的心肌细胞和来自急性心肌梗死（AMI）小鼠模型的心脏样品中定量miR-199 a-5 p表达水平。miR-199 a-5 p过表达用于确定其在QL对心肌细胞的抗肥大作用中的作用。结果如下：PE可使心肌细胞表面明显增大，ANP、BNP、MYH 7 mRNA水平升高，蛋白/DNA比值升高。所有这些变化都被QL治疗逆转。同时，miR-199 a-5 p在AMI小鼠心肌组织中表达增加。值得注意的是，PE处理的心肌细胞中miR-199 a-5 p的增加被QL处理逆转。此外，miR-199 a-5 p的过表达取消了QL对心肌细胞的抗肥大作用。结论：QL对PE诱导的心肌肥厚有保护作用。miR-199 a-5 p在心脏肥大中增加，而通过用QL处理降低。miR-199 a-5 p抑制是QL对心肌细胞的抗肥大作用所必需的。
Background/Aims: Qiliqiangxin (QL), a traditional Chinese medicine, has long been used to treat chronic heart failure. Previous studies demonstrated that QL could prevent cardiac remodeling and hypertrophy in response to hypertensive or ischemic stress. However, little is known about whether QL could modulate cardiac hypertrophy in vitro, and (if so) whether it is through modulation of specific hypertrophy-related microRNA. Methods: The primary neonatal rat ventricular cardiomyocytes were isolated, cultured, and treated with phenylephrine (PE, 50 µmol/L, 48 h) to induce hypertrophy in vitro, in the presence or absence of pretreatment with QL (0.5 µg/ml, 48 h). The cell surface area was determined by immunofluorescent staining for α-actinin. The mRNA levels of hypertrophic markers including atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and β-myosin heavy chain (MYH7) were assayed by qRT-PCRs. The protein synthesis of cardiomyocytes was determined by the protein/DNA ratio. The miR-199a-5p expression level was quantified in PE-treated cardiomyocytes and heart samples from acute myocardial infarction (AMI) mouse model. MiR-199a-5p overexpression was used to determine its role in the anti-hypertrophic effect of QL on cardiomyocytes. Results: PE induced obvious enlargement of cell surface in cardiomyocytes, paralleling with increased ANP, BNP, and MYH7 mRNA levels and elevated protein/DNA ratio. All these changes were reversed by the treatment with QL. Meanwhile, miR-199a-5p was increased in AMI mouse heart tissues. Of note, the increase of miR-199a-5p in PE-treated cardiomyocytes was reversed by the treatment with QL. Moreover, overexpression of miR-199a-5p abolished the anti-hypertrophic effect of QL on cardiomyocytes. Conclusion: QL prevents PE-induced cardiac hypertrophy. MiR-199a-5p is increased in cardiac hypertrophy, while reduced by treatment with QL. miR-199a-5p suppression is essential for the anti-hypertrophic effect of QL on cardiomyocytes.