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目的:研究芒果苷对柔红霉素致心肌细胞凋亡的拮抗作用及其可能机制。方法:取原代培养心肌细胞以1μmol/L柔红霉素(daunorubicin,DNR)建立心肌细胞凋亡模型,以不同浓度芒果苷(25、50、100μmol/L)干预,选择银杏叶提取物EGB761(25μg/mL)作为阳性对照药物。应用光学倒置显微镜及电子显微镜观察凋亡细胞形态改变,用流式细胞术(Annexin V–FITC/PI双染色法)检测细胞凋亡率,用分光光度法检测细胞裂解液中Caspase-3酶活性,用实时荧光定量RT-PCR方法检测P53、Fas及FasL表达水平。结果:芒果苷作用于柔红霉素诱导的心肌细胞凋亡模型后,能明显抑制柔红霉素诱导的心肌细胞凋亡,降低心肌细胞凋亡率,下调Fas和FasL表达、抑制Caspase-3活化,随着药物浓度增加,抑制凋亡作用增强,其作用强度与芒果苷浓度有关,呈剂量依赖性。结论:芒果苷能有效抑制柔红霉素诱导的心肌细胞凋亡,其作用机制可能与下调Fas和FasL表达水平及抑制Caspase-3活化有关。
Objective: To study the antagonistic effect of mangiferin on daunomycin-induced cardiomyocyte apoptosis and its possible mechanism. Methods: Myocardial cell apoptosis model was established by primary cultured cardiomyocytes with 1μmol / L daunorubicin (DNR). The effects of different concentrations of mangiferin (25, 50, 100μmol / L) (25 μg / mL) as a positive control drug. Morphological changes of apoptotic cells were observed with optical inverted microscope and electron microscope. Apoptosis rate was detected by flow cytometry (Annexin V-FITC / PI double staining), and the activity of Caspase-3 in cell lysate was detected by spectrophotometry The expression of P53, Fas and FasL were detected by real-time fluorescence quantitative RT-PCR. Results: Mangiferin inhibited daunomycin-induced cardiomyocyte apoptosis, decreased cardiomyocyte apoptosis, decreased Fas and FasL expression, and inhibited Caspase-3 after daunorubicin-induced cardiomyocyte apoptosis model Activation, with the drug concentration increases, the role of inhibition of apoptosis increased, the role of strength and the concentration of mangiferin in a dose-dependent manner. CONCLUSION: Mangiferin can effectively inhibit daunomycin-induced cardiomyocyte apoptosis. Its mechanism may be related to the down-regulation of Fas and FasL expression and the inhibition of Caspase-3 activation.