| 沉默BHLHE41对血管紧张素Ⅱ诱导的 H9C2心肌细胞肥大和凋亡的影响 |
| 投稿时间：2024-09-18 修订日期：2024-10-09 点此下载全文 |
| 引用本文：徐路晴,余娜,江克兵,沈涤非.沉默BHLHE41对血管紧张素Ⅱ诱导的 H9C2心肌细胞肥大和凋亡的影响[J].医学研究杂志,2025,54(2):70-76 |
| DOI：
10.11969/j.issn.1673-548X.2025.02.012 |
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| 基金项目:国家自然科学基金资助项目(81660039) |
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| 中文摘要:目的 探究基本螺旋-环-螺旋家族成员e41(basic helix-loop-helix family member e41, BHLHE41)对血管紧张素Ⅱ(angiotensin Ⅱ,AngⅡ)刺激下的H9C2大鼠心肌细胞肥大和凋亡的影响。方法 为探讨AngⅡ刺激下的BHLHE41的表达水平,将细胞分为两组,即正常组和AngⅡ组。为了探究沉默BHLHE41(silencing BHLHE41, siBHLHE41)对心肌细胞肥大与凋亡的影响,将心肌细胞分为4组,即空白组[转染小干扰RNA(siRNA)+磷酸盐缓冲盐水(phosphate buffered saline,PBS)]、siBHLHE41组(转染siBHLHE41+PBS)、模型组(转染siRNA+AngⅡ)和实验组(转染siBHLHE41+AngⅡ)。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction, RT-qPCR)法检测BHLHE41、β-肌球蛋白重链(β-myosin heavy chain, β-MHC)、心房钠尿肽(atrial natriuretic peptide, ANP)、B型钠尿肽(B-type natriuretic peptide, BNP)、聚腺苷二磷酸核糖聚合酶(poly ADP-ribose polymerase, PARP)、B淋巴细胞瘤2基因(B-cell lymphoma-2, Bcl-2)、B细胞淋巴瘤-2相关X蛋白(Bcl-2-associated X protein, Bax)和p53的 mRNA表达水平。采用Western blot法检测BHLHE41、Bax、Bcl-2、PARP及p53的蛋白表达水平。鬼笔环肽染色观察心肌细胞表面积,原位末端脱氧核苷酸转移酶标记法(terminal deoxynucleotidyl trann sferase-mediated dUTP-biotin nick end labeling,TUNEL)观察心肌细胞凋亡情况。结果 与正常组比较,AngⅡ组BHLHE41mRNA水平及蛋白表达水平均升高。与空白组比较,模型组β-MHC、ANP、BNP mRNA表达水平上调,PARP、Bax、p53 蛋白和mRNA表达水平明显升高,Bcl-2蛋白及mRNA表达水平明显降低,心肌细胞表面积明显增大,心肌细胞凋亡率显著升高(P＜0.05)；与模型组比较,实验组β-MHC、ANP、BNP mRNA表达水平上升,PARP、Bax、p53蛋白和mRNA表达水平明显升高,Bcl-2蛋白及mRNA表达水平明显降低,心肌细胞表面积增大,心肌细胞凋亡率显著升高(P＜0.05)。结论 沉默BHLHE41可以加重AngⅡ诱导的H9C2心肌细胞肥大与凋亡,这可能与凋亡相关信号通路p53激活有关。 |
| 中文关键词:心肌细胞 肥大 细胞凋亡 BHLHE41 大鼠 |
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| Effect of Silencing BHLHE41 on Angiotensin II-induced Hypertrophy and Apoptosis of H9C2 Cardiomyocytes. |
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| Abstract:Objective To investigate the effects of basic helix-loop-helix family member e41 (BHLHE41) on angiotensin Ⅱ (AngⅡ)-induced hypertrophy and apoptosis of H9C2 cardiomyocytes. Methods The expression level of BHLHE41stimulated by AngⅡ was investigated, and the cells were divided into normal group and AngⅡ group. To investigate the effect of silencing BHLHE41 (siBHLHE41) on cardiomyocyte hypertrophy and apoptosis, the cells were divided into 4groups:blank group [transfected with small interfering RNA (siRNA)+ phosphate buffered saline (PBS)], siBHLHE41group [transfected with siBHLHE41 +PBS], model group (transfected with siRNA+ AngⅡ) and experimental group (transfected with siBHLHE41+ AngⅡ). The mRNA levels of BHLHE41, β-myosin heavy chain (β-MHC), atrial natriuretic peptide (ANP), B-type natriuretic peptide (BNP), poly ADP-ribose polymerase (PARP), B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-2-associated X protein (Bax), and p53 were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The protein expression levels of BHLHE41, Bax, Bcl-2, PARP and p53 were detected by Western blot. Phalloidin staining was used to observe the surface area of cardiomyocytes, TUNEL staining was used to observe cardiomyocytes apoptosis. Results Compared with the normal group, the mRNA and protein expression levels of BHLHE41 was higher in the AngⅡ group. Compared with blank group, the mRNA expression levels of β-MHC, ANP, BNP in the model group were upregulated, and the protein and mRNA expression levels of PARP, Bax and p53 in model group were significantly increased, the expression level of Bcl-2 was significantly decreased, the surface area of cardiomyocytes was significantly increased, and the apoptosis rate of cardiomyocytes was significantly increased (P＜0.05). Compared with model group, the mRNA expression levels of β-MHC, ANP and BNP in the experimental group were upregulated, and the protein and mRNA expression levels of PARP, Bax and p53 in experimental group were significantly increased, the expression level of Bcl-2 was significantly decreased, the surface area of cardiomyocytes was increased, and the apoptosis rate of cardiomyocytes was significantly increased (P＜0.05). Conclusion Silencing BHLHE41 can aggravate AngⅡ-induced hypertrophy and apoptosis of H9C2 cardiomyocytes, which may be related to the p53 activation of the apoptotic signaling pathway. |
| keywords:Cardiomyocytes Hypertrophy Apoptosis BHLHE41 Rats |
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