从钙超载角度探讨通脉养心丸心肌保护作用的机制(1)
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摘要：目的从抑制钙超载的角度探讨通脉养心丸抗细胞凋亡保护心肌的作用机制。方法采用大鼠心肌细胞原代培养方法获得正常心肌细胞并建立缺氧损伤细胞模型，采用FLIPR Calcium 4 试剂盒检测药物对胞浆内钙离子浓度变化的影响；Annexin V/PI双染法检测心肌细胞总凋亡率。结果缺氧损伤组心肌细胞胞浆内钙离子浓度显著升高(P<0.01)，通脉养心丸各剂量组变化不显著；再次加药后，通脉养心丸各剂量组胞浆内钙离子浓度均显著低于缺氧损伤组(P<0.01)。缺氧损伤组心肌细胞凋亡率显著升高(P<0.01)，通脉养心丸4 g/kg、8 g/kg剂量组可显著降低心肌细胞凋亡率(P<0.01)。结论抑制心肌细胞钙超载是通脉养心丸抗心肌细胞凋亡保护心肌的作用机制之一。
关键词：通脉养心丸 心肌细胞 钙超载 细胞凋亡
中图分类号：R542.2R289.5文献标识码：A文章编号：1672-1349(2011)05-0562-02
Investigation of Protection Mechanism of Tongmai Yangxin Pill on Myocardium from the Perspective of Calcium Overload
Xiao Yang，Zhang Jiafu，Zhang Ling，et al
Tianjin University of Traditional Chinese Medicine (Tianjin 300193)
Abstract:ObjectiveTo observe the effect of Tongmai Yangxin pill (TYP) on apoptosis and [Ca2+]i in cultured hypoxia cardiomyocytes，and explore the mechanism of myocardial protection.MethodsPrimarily cultured neonate rat cardiomyocytes were used to establish the hypoxia models.[Ca2+]i of cardiomyocytes loaded by Fluo-4 was analyzed by Multiskan Ascent immediately.The apoptosis percentage of cardiomyocytes was measured by Annexin V/PI flow cytometry.ResultsCalcium in hypoxia cardiac myocytes injury group was significantly higher. Calcium in TYP group were significantly lower than that in hypoxic injury group(P<0.01).Myocardial hypoxia significantly increased apoptosis rate(P<0.01).The apoptosis rate was significantly reduce in 4 g/kg TYP group and 8 g/kg TYP group(P<0.01).ConclusionTYP could inhibit calcium overload in myocardial cells，thereby to reduce the incidence of myocardial apoptosis.
Key words：Tongmai Yangxin pill；Cardiomyocytes；Calcium overload；apoptosis
通脉养心丸是由传统组方生脉散与炙甘草汤加减而成，具有养心补血、通脉止痛的功效。临床主要用于胸痹心痛、心悸怔忡等证的治疗。长期临床实践表明，通脉养心丸可有效缓解胸痛、胸闷憋气、心悸气短等症状，减少心绞痛发作频数。但对其药理作用机制的研究尚不完善。本研究采用大鼠心肌细胞原代培养方法获得实验用正常心肌细胞，并建立缺氧损伤细胞模型，从抑制心肌细胞钙超载的角度探讨通脉养心丸抗细胞凋亡，保护心肌的作用机制，为临床应用提供科学依据。
1材料与方法
1.1实验动物Wistar大鼠，50只，体重300 g±20 g，雄性，购于中国医学科学院协和动物研究所。Wistar大鼠，20只，1 d~3 d龄，雌雄不拘，购于中国医学科学院放射研究所。
1.2药品与试剂通脉养心丸(天津中新药业集团股份有限公司乐仁堂制药厂提供，每10丸重1 g,C107074)；DMEM/F12培养基(Hyclone，NUH0180)；胰蛋白酶(1:250 ......
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