加味丹参饮对大鼠心肌缺血再灌注损伤SSAT活性的影响
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摘要
目的观察加味丹参饮对SD大鼠心肌缺血再灌注损伤(IRI)模型精脒/精胺乙酰转移酶(SSAT)/多胺通路的影响,探讨其抗IRI的机制。方法结扎大鼠冠状动脉左前降支30 min再灌注90 min建立大鼠心肌IRI模型。实验大鼠随机分为正常对照组、假手术组、模型组和加味丹参饮组,每组10只。TTC染色测定大鼠心肌梗死面积,ELISA检测大鼠心肌组织SSAT含量,RT-PCR和Western blot分别检测大鼠心肌组织SSAT m RNA和蛋白表达,HPLC检测大鼠心肌组织多胺(腐胺、精脒、精胺)含量。结果与假手术组比较,模型组大鼠心肌梗死面积、SSAT含量增加,SSAT m RNA和蛋白表达升高,多胺含量降低(P<0.05,P<0.01);与模型组比较,加味丹参饮组大鼠心肌梗死面积、SSAT含量减少,SSAT m RNA和蛋白表达降低,多胺含量升高(P<0.05,P<0.01)。结论加味丹参饮可能通过调节SSAT/多胺通路,增加心肌细胞多胺含量,从而对大鼠心肌IRI发挥保护作用。
Objective To observe the effects of modified Danshen Decoction on spermidine/spermine acetyltransferase(SSAT)/polyamine pathways of SD rats with IRI; To investigate its protective mechanism. Methods The model of IRI was established by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 90 min. The SD rats were randomly divided into the control group, sham-operation group, model group and modified Danshen Decoction group, with 10 rats in each group. The myocardial infarction size was measured by using TTC staining. The contents of SSAT were measured by ELISA. The SSAT m RNA and SSAT protein expression level were detected with real-time fluorescent quantitative PCR method and Western blot, respectively. The contents of polyamines(putrescine, spermidine, spermine) in cardiac tissue were detected by HPLC. Results Compared with sham-operation group, the myocardial infarction size, the SSAT content, the SSAT m RNA and SSAT protein expression levels of model group increased significantly, the contents of polyamines decreased significantly, with statistical significance(P<0.01); Compared with model group, the myocardial infarction size of modified Danshen Decoction group was significantly reduced, while the SSAT content and SSAT m RNA and protein expression level decreased significantly, the contents of polyamines increased, with statistical significance(P<0.05, P<0.01). Conclusion Modified Danshen Decoction can adjust the SSAT polyamine pathways and increase polyamine content in cardiomyocytes, and thus play a role of protection of myocardial ischemia-reperfusion injury.
Objective To observe the effects of modified Danshen Decoction on spermidine/spermine acetyltransferase(SSAT)/polyamine pathways of SD rats with IRI; To investigate its protective mechanism. Methods The model of IRI was established by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 90 min. The SD rats were randomly divided into the control group, sham-operation group, model group and modified Danshen Decoction group, with 10 rats in each group. The myocardial infarction size was measured by using TTC staining. The contents of SSAT were measured by ELISA. The SSAT m RNA and SSAT protein expression level were detected with real-time fluorescent quantitative PCR method and Western blot, respectively. The contents of polyamines(putrescine, spermidine, spermine) in cardiac tissue were detected by HPLC. Results Compared with sham-operation group, the myocardial infarction size, the SSAT content, the SSAT m RNA and SSAT protein expression levels of model group increased significantly, the contents of polyamines decreased significantly, with statistical significance(P<0.01); Compared with model group, the myocardial infarction size of modified Danshen Decoction group was significantly reduced, while the SSAT content and SSAT m RNA and protein expression level decreased significantly, the contents of polyamines increased, with statistical significance(P<0.05, P<0.01). Conclusion Modified Danshen Decoction can adjust the SSAT polyamine pathways and increase polyamine content in cardiomyocytes, and thus play a role of protection of myocardial ischemia-reperfusion injury.
引文
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[6]李鑫辉,黄政德,葛金文.加味丹参饮对血瘀证兔心肌缺血再灌注损伤的影响[J].中国中医药信息杂志,2011,18(6):37-39.
[7]潘小平,黄政德,杨伟峰,等.加味丹参饮对动脉粥样硬化模型小鼠内皮细胞功能及超微结构的影响[J].中国中医药信息杂志,2015,22(11):42-45.
[8]彭霞,黄政德,成细华,等.加味丹参饮预处理调控自噬相关基因Beclin1和Atg5表达抗大鼠心肌缺血再灌注损伤[J].湖南中医药大学学报,2016,36(7):20-24.
[9]赵雅君,徐长庆,李全风,等.应用反向高效液相色谱分析离体心脏灌流大鼠心肌组织中的多胺[J].中国病理生理杂志,2005,21(2):412-413.
[10]赵雅君,王丽娜,李鸿珠,等.大鼠心肌多胺代谢限速酶ODC、SSAT活性分析[J].中国药理学通报,2005,21(5):555-559.
[11]CERRADA-GIMENZ M,PIETIL?M,LOIMAS S,el a1.Continuous oxidative stress due to activation of polyamine catabolism accelerates aging and protects against hepatotoxic insults[J].Transgenic Res,2011,20:387-396.
[12]林岩,肖薇,李波.二氟甲基鸟氨酸对异丙肾上腺素诱导的心肌细胞肥大与凋亡的影响[J].中国药理学通报,2015,31(5):668-672.
[2]DING L,BA X H.Role of ornithine decarboxylase/polyamine pathway in focal cerebral ischemia-reperfusion injury and its mechanism in rats[J].Int J Clin Exp Med,2015,8(11):20624-20630.
[3]韩丽萍,徐长庆,姜春明,等.多胺代谢与大鼠心肌缺血再灌注损伤关系的实验研究[J].中华心血管病杂志,2008,36(4):346-349.
[4]RYU J H,CHO Y S,CHUN Y S,et al.Myocardial SSAT induction via AMPK signaling and its implication for ischemic injury[J].Biochem Biophys Res Commum,2008,366(2):438-444.
[5]王庆高,黄政德,肖健,等.加味丹参饮预处理对乳鼠缺氧/复氧心肌细胞的延迟保护作用及对蛋白激酶C的影响[J].中西医结合心脑血管病杂志,2007,5(10):953-955.
[6]李鑫辉,黄政德,葛金文.加味丹参饮对血瘀证兔心肌缺血再灌注损伤的影响[J].中国中医药信息杂志,2011,18(6):37-39.
[7]潘小平,黄政德,杨伟峰,等.加味丹参饮对动脉粥样硬化模型小鼠内皮细胞功能及超微结构的影响[J].中国中医药信息杂志,2015,22(11):42-45.
[8]彭霞,黄政德,成细华,等.加味丹参饮预处理调控自噬相关基因Beclin1和Atg5表达抗大鼠心肌缺血再灌注损伤[J].湖南中医药大学学报,2016,36(7):20-24.
[9]赵雅君,徐长庆,李全风,等.应用反向高效液相色谱分析离体心脏灌流大鼠心肌组织中的多胺[J].中国病理生理杂志,2005,21(2):412-413.
[10]赵雅君,王丽娜,李鸿珠,等.大鼠心肌多胺代谢限速酶ODC、SSAT活性分析[J].中国药理学通报,2005,21(5):555-559.
[11]CERRADA-GIMENZ M,PIETIL?M,LOIMAS S,el a1.Continuous oxidative stress due to activation of polyamine catabolism accelerates aging and protects against hepatotoxic insults[J].Transgenic Res,2011,20:387-396.
[12]林岩,肖薇,李波.二氟甲基鸟氨酸对异丙肾上腺素诱导的心肌细胞肥大与凋亡的影响[J].中国药理学通报,2015,31(5):668-672.