黄芪甲苷对D-半乳糖诱导原代培养心肌细胞凋亡的保护作用及机制研究
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篇名: 黄芪甲苷对D-半乳糖诱导原代培养心肌细胞凋亡的保护作用及机制研究
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摘要: 目的:研究黄芪甲苷对D-半乳糖(D-gal)诱导原代培养心肌细胞凋亡的保护作用及机制。方法:分离和培养Wistar乳鼠原代心肌细胞,将细胞分为正常对照组(无血清DMEM高糖培养基)、D-gal组(含5 g/L D-gal的无血清DMEM高糖培养基)和黄芪甲苷低、中、高质量浓度组(分别先以含25、50、100 μg/mL黄芪甲苷的无血清DMEM高糖培养基培养1 h,然后换为含有相应质量浓度黄芪甲苷和5 g/L D-gal的无血清DMEM高糖培养基)。采用CCK-8法检测培养48 h后细胞活性,分别采用Hoechst染色法(培养24 h)和流式细胞术(培养48 h)考察细胞凋亡情况,采用实时荧光定量-聚合酶链式反应法检测培养24 h后细胞中凋亡相关因子(Bcl-2、Bax、Caspase-3)和心房钠尿肽(ANP) mRNA表达水平。结果:与正常对照组比较,D-gal组细胞的活性以及细胞中Bcl-2 mRNA水平、Bcl-2/Bax比值降低,细胞凋亡率和细胞中Bax、Caspase-3、ANP mRNA水平升高,差异均有统计学意义(P<0.05或P<0.01)。与D-gal组比较,黄芪甲苷各浓度组细胞的活性以及细胞中Bcl-2 mRNA水平、Bcl-2/Bax比值均升高,细胞凋亡率和细胞中Bax、Caspase-3、ANP mRNA水平均降低,差异均有统计学意义(P<0.05或P<0.01),且具有一定浓度依赖性。结论:黄芪甲苷对D-gal诱导的原代心肌细胞凋亡具有一定的保护作用,其机制可能与升高细胞中Bcl-2/Bax比值和下调细胞中Caspase-3、ANP mRNA的表达有关。
ABSTRACT: OBJECTIVE: To study the protective effect and mechanism of astragaloside Ⅳ on D-galactose (D-gal)-induced primary cardiomyocytes apoptosis. METHODS: Wistar neonatal rat primary cardiomyocytes were isolated and cultured. The cardiomyocytes were divided into normal control group (DMEM high glucose medium without serum), D-gal group (DMEM high glucose medium without serum containing 5 g/L D-gal) and astragaloside Ⅳ low-concentration, medium-concentration and high-concentration groups (after cultured with DMEM high glucose medium without serum containing 25, 50, 100 μg/mL astragaloside Ⅳ for 1 h, and then replaced with DMEM high glucose medium without serum containing corresponding concentration of astragaloside Ⅳ and 5 g/L D-gal). Cardiomyocytes activity was detected by CCK-8 kit after cultured for 48 h. The apoptosis level was detected by Hoechst staining (cultured for 24 h) and flow cytometry (cultured for 48 h). The mRNA expressions of apoptosis related factors (Bcl-2, Bax, Caspase-3) and ANP in cardiomyocytes were detected by RT-PCR after cultured for 24 h. RESULTS: Compared with normal control group, the activity of cardiomyocytes, mRNA level of Bcl-2 and ratio of Bcl-2/Bax were decreased in D-gal group, while apoptosis rate and mRNA levels of Bax, Caspase-3 and ANP were increased, with statistical significance (P<0.05 or P<0.01). Compared with D-gal group, the activity of cardiomyocytes, mRNA level of Bcl-2 and ratio of Bcl-2/Bax were increased in astragaloside Ⅳ groups, while apoptosis rate and mRNA levels of Bax, Caspase-3 and ANP were decreased, with statistical significance (P<0.05 or P<0.01), and in concentration-dependent manner. CONCLUSIONS: Astragaloside Ⅳ can protect D-gal induced primary cardiomyocytes from apoptosis, the mechanism of which may be associated with the increase of Bcl-2/Bax ratio and the decrease of mRNA expressions of Caspase-3 and ANP.
期刊: 2018年第29卷第9期
作者: 王晓霞,刘天龙,刘晶,刘小玲,张勇,肖云峰,刘小雷
AUTHORS: WANG Xiaoxia,LIU Tianlong,LIU Jing,LIU Xiaoling,ZHANG Yong,XIAO Yunfeng,LIU Xiaolei
关键字: 黄芪甲苷;D-半乳糖;原代心肌细胞;凋亡;保护作用
KEYWORDS: Astragaloside Ⅳ; D-galactose; Primary cardiomyocytes; Apoptosis; Protective effect
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