加参方浸膏对H2O2诱导的H9c2心肌细胞凋亡的干预作用研究-中国药房

加参方浸膏对H2O2诱导的H9c2心肌细胞凋亡的干预作用研究

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篇名：	加参方浸膏对H2O2诱导的H9c2心肌细胞凋亡的干预作用研究
TITLE：
摘要：	目的：观察加参方浸膏对过氧化氢（H2O2）诱导的H9c2心肌细胞凋亡的干预作用，并探讨其作用机制。方法：采用MTT法，考察400、200、100、50、25 μmol/L的H2O2与3.0、1.8、0.9、0.3、0.1 mg/mL加参方浸膏对H9c2心肌细胞活性的影响。将H9c2心肌细胞分为正常组、模型组、加参方组，除正常组细胞加入相应培养基外，模型组、加参方组细胞均采用50 μmol/L H2O2处理以诱导细胞发生氧化应激损伤，加参方组在模型组基础上加入0.3 mg/mL加参方浸膏干预处理。采用DAPI染色法，在荧光显微镜下观察细胞形态学变化；采用流式细胞仪检测细胞凋亡率；采用荧光定量逆转录-聚合酶链式反应法测定细胞中胱天蛋白酶3（Caspase-3）、Bax、Bcl-2 mRNA表达水平，Western Blot法测定Caspase-3、Bax、Bcl-2蛋白表达水平。结果：50 μmol/L为H2O2最适造模浓度，0.3 mg/mL为加参方浸膏最适药物干预浓度。与模型组比较，加参方组细胞的形态改善，细胞凋亡率显著降低（P＜0.05）；Caspase-3、Bax mRNA及蛋白表达水平显著降低，Bcl-2 mRNA及蛋白表达水平显著升高（P＜0.05）。结论：加参方浸膏对H2O2诱导的H9c2心肌细胞凋亡有显著的抑制作用，其机制可能与下调细胞中Caspase-3、Bax表达，上调Bcl-2表达有关。
ABSTRACT：	OBJECTIVE： To observe the intervention effects of Jiashen formula extractum on hydrogen peroxide（H2O2）-induced apoptosis of H9c2 cardiomyocytes， and to investigate its mechanism. METHODS： MTT assay was used to investigate the effects of 400， 200， 100， 50， 25 μmol/L H2O2 and 3.0， 1.8， 0.9， 0.3， 0.1 mg/mL Jiashen formula extractum on the activity of H9c2 cardiomyocytes. H9c2 cardiomyocytes were divided into normal group， model group and Jiashen formula group. Model group and Jiashen formula group were given 50 μmol/L H2O2 to induce oxidative stress injury except that normal group was cultured with corresponding culture medium. Jiashen formula group was given 0.3 mg/mL Jiashen formula extractum for intervention on the basis of model group. Cell morphology was observed by fluorescence microscope through DAPI dye. The apoptosis rate was detected by flow cytometry. Fluorescence quantitative RT-PCR was used to detect mRNA expression levels of Caspase-3， Bax and Bcl-2 in cells； Western Blot assay was used to determine the protein expression levels of Caspase-3， Bax and Bcl-2. RESULTS： 50 μmol/L was the most suitable modeling concentration of H2O2； 0.3 mg/mL was the most suitable intervention concentration of Jiashen formula extractum. Compared with model group， cells in Jiashen formula extractum group were significantly improved in morphology and reduced in apoptosis rate； mRNA and protein expression levels of Caspase-3 and Bax were significantly reduced， and those of Bcl-2 were significantly increased （P＜0.05）. CONCLUSIONS： Jiashen formula extractum shows significant inhibitory effect on H2O2-induced apoptosis of H9c2 cardiomyocytes， the mechanism of which may be associated with down-regulation of Caspase-3 and Bax expression and up-regulation of Bcl-2 expression in cardiomyocytes.
期刊：	2018年第29卷第14期
作者：	郝轩轩，王新陆，崔琳，王幼平，李彬，谢世阳，高原，朱明军
AUTHORS：	HAO Xuanxuan，WANG Xinlu，CUI Lin，WANG Youping，LI Bin，XIE Shiyang，GAO Yuan，ZHU Mingjun
关键字：	加参方；浸膏；过氧化氢；H9c2心肌细胞；凋亡；胱天蛋白酶3；Bax；Bcl-2
KEYWORDS：	Jiashen formula； Extractum； Hydrogen peroxide； H9c2 cardiomyocytes； Apoptosis； Caspase-3； Bax； Bcl-2
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