| 血管紧张素Ⅱ诱导的新生小鼠心肌细胞肥大模型 |
| 投稿时间：2015-04-06 修订日期：2015-04-10 点此下载全文 |
| 引用本文：周恒,唐其柱,邓伟,卞洲艳,袁园,倪健.血管紧张素Ⅱ诱导的新生小鼠心肌细胞肥大模型[J].医学研究杂志,2015,44(11):26-30 |
| DOI：
10.11969/j.issn.1673-548X.2015.11.008 |
| 摘要点击次数: 1077 |
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| 基金项目:国家自然科学基金资助项目(81300070、81270303、81470516);教育部高等学校博士学科点专项科研基金资助项目(20130141130010) |
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| 中文摘要:目的 探索新生小鼠心肌细胞的原代培养方法,建立血管紧张素Ⅱ(AngⅡ)诱导的小鼠心肌细胞肥大模型。方法 使用0.03%的胰酶+0.04%的2型胶原酶多次消化分离心肌细胞,差时贴壁法纯化心肌细胞。1μmol/L AngⅡ刺激心肌细胞肥大,α-actinin染色进行心肌细胞纯度鉴定及面积检测,实时定量RT-PCR检测心肌细胞肥大标志物的表达,Western blot法检测蛋白质磷酸化水平。结果 培养得到的小鼠心肌细胞具有较高纯度及存活力;AngⅡ刺激后心肌细胞面积增大、蛋白合成增加,ANP、BNP、β-MHC等心肌细胞肥大标志物的mRNA表达水平较对照组明显升高(P<0.05),而α-MHC表达则下降(P<0.05);此外,AngⅡ刺激组ERK、JNK与p38的磷酸化水平较对照组明显上调(P<0.05)。结论 使用改良的新生小鼠心肌细胞原代培养方法,成功培养了具有较高纯度及存活力的小鼠心肌细胞,并建立了AngⅡ诱导的小鼠心肌细胞肥大模型。 |
| 中文关键词:心肌细胞 原代培养 血管紧张素Ⅱ 肥大 丝裂原活化蛋白激酶 |
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| AngiotensinⅡ Induced Hypertrophy of Neonatal Mouse Cardiomyocytes. |
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| Abstract:Objective To explore the protocol of primary culture of neonatal mouse cardiomyocytes and establish angiotensinⅡ-induced hypertrophic model. Methods The cardiomyocytes were isolated by 0.03% trypsin and 0.04% collagenase type Ⅱ digestion, and purified by differential attachment technique. Cardiomyocyte hypertrophy was induced using 1mol/L angiotensinⅡ, areas of cardiomyocytes were detected using α-actinin staining, expression of hypertrophic markers were measured by real-time quantitative RT-PCR, and protein phosphorylations were measured by Western blot. Results Cultured mouse cardiomyocytes exhibited high purity and viability. Angiotensin Ⅱ induced increases in cell area, protein synthesis, and expression levels of ANP, BNP and β-MHC, and decrease in expression level of α-MHC compared with those in control group(P<0.05). In addition, angiotensin Ⅱ significantly increased the phosphorylation levels of ERK, JNK and p38 compared with control group(P<0.05). Conclusion We obtained neonatal mouse cardiomyocytes with high purity and viability by using the modified method of primary culture, and established angiotensin Ⅱ-induced hypertrophic model of mouse cardiomyocytes. |
| keywords:Cardiomyocyte Primary culture Angiotensin Ⅱ MAPKs |
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