黄芪甲苷对阿霉素诱导的HL-1细胞凋亡的抑制作用及机制研究

贾婷; 崔梁瑜; 祁玉营; 刘欢; 薛松妍; 王方圆; 马静

doi:10.12125/j.chj.202306065

黄芪甲苷对阿霉素诱导的HL-1细胞凋亡的抑制作用及机制研究

Protective effect of astragaloside IV on adriamycin-induced HL-1 cell injury and its mechanism

摘要

摘要:
目的探讨黄芪甲苷（astragaloside IV，ASIV）对阿霉素诱导HL-1细胞凋亡的影响及机制，为进一步研发治疗阿霉素诱导心肌损伤的新药提供可靠的实验依据。
方法将HL-1 心肌细胞分为正常对照组（Control组）、模型组（Model组）、1 μmol/L ASIV 、2 μmol/L ASIV 及4 μmol/L ASIV 组等5组。用 5.2 μmol/L的 DOX 溶液放入细胞培养箱中培养 12 h制备心肌细胞损伤模型后，以1 μmol/L、2 μmol/L、4 μmol/L 的 ASIV 溶液加入细胞损伤模型中，再放入细胞培养箱中培养12 h。CCK8 法确定阿霉素造模药物浓度，流式细胞术确定浓度阿霉素造模时间，荧光染色法检测不同浓度ASIV对阿霉素诱导的HL-1心肌细胞凋亡、线粒体膜电位水平及ROS水平；分光光度检测法检测ASIV 对阿霉素诱导的HL-1 心肌细胞上清NO含量。
结果 5.2 μmol/L的DOX 培养12 h诱导HL-1心肌细胞凋亡模型。与Model组相比，2 μmol/L 和4 μmol/L ASIV组细胞凋亡明显减少（P<0.01），NO含量显著增加（P<0.01）；ASIV各剂量组Annexin V-FITC表达均有降低趋势（P<0.01）、ASIV 1 μmol/L、ASIV 2 μmol/L Mito-Tracker Red CMXRos表达均明显升高（P<0.05），ASIV 4 μmol/L Mito-Tracker Red CMXRos表达均明显升高（P<0.01），ASIV 1 μmol/L、ASIV 2 μmol/L和ASIV 4 μmol/L组ROS表达显著降低（P<0.05）。
结论 ASIV可通过减轻心肌细胞膜电位下降，降低ROS 表达，提高 NO含量，减轻氧化应激损伤，从而抑制阿霉素诱导的心肌细胞凋亡，且呈剂量依赖性。

Abstract:
AIM To investigate the protective effect of astragaloside IV (ASIV) on doxorubicin-induced HL-1 cell injury and its related mechanism, and to provide cardiac therapy for doxorubicin-induced myocardial injury.
METHODS HL-1 cardiomyocytes were divided into normal control group, model group, 1 μmol/L ASIV group, 2 μmol/L ASIV group and 4 μmol/L ASIV group. The myocardial cell injury model was made with DOX solution of 5.2 μmol/L and cultured in a cell incubator for 12h. ASIV solutions of 1 μmol/L, 2 μmol/L and 4 μmol/L were added into the culture bottles of the corresponding groups and then they were placed into the cell culture box for culture. After 12 hours, the ASIV solutions were removed to detect the relevant indexes. CCK8 method was used to determine the concentration of adriamycin modeling drug, flow cytometry was used to observe the concentration of adriamycin modeling time, and fluorescence staining was used to detect the level of apoptosis, mitochondrial membrane potential and ROS of adriamycin induced HL-1 cardiomyocytes by ASIV. The content of NO in the supernatant of adriamycin induced HL-1 cardiomyocytes was detected by spectrophotometry.
RESULTS The model concentration of adriamycin was 5.2 μmol/L. The apoptosis time of HL-1 myocardial cells induced by adriamycin was 12h. After adriamycin administration, apoptosis was significantly increased in model group (P<0.01) and it significantly decreased in 2 μmol/L and 4 μmol/L ASIV groups (P<0.01). Compared with those in model group, ROS expression was decreased and NO concentration and mitochondrial intima potential were increased in ASIV treatment groups. In 4 μmol/L ASIV group, ROS concentration was significantly decreased, while NO concentration and mitochondrial intima potential were significantly increased (P<0.05).
CONCLUSION ASIV may inhibit adriamycin induced cardiomyocyte apoptosis in a dose-dependent manner by reducing the decline of membrane potential of cardiomyocytes, reducing ROS expression, increasing NO content, and alleviating oxidative stress injury.

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