补阳还五汤对舒张性心衰大鼠心肌线粒体能量代谢及AMPK/PPARα信号通路的影响
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摘要
目的:探讨补阳还五汤基于腺苷酸激活蛋白激酶(AMPK)/过氧化物酶体增殖物激活受体α(PPARα)信号通路对舒张性心衰(DHF)大鼠心肌线粒体能量代谢的影响及其机制研究。方法:将48只SD大鼠随机分为假手术组与模型组,模型组大鼠运用腹主动脉缩窄法建立DHF大鼠模型。将造模成功的大鼠随机分为模型组,补阳还五汤组(12.72 g·kg~(-1)·d~(-1)),酒石酸美托洛尔组(0.004 5 g·kg~(-1)·d~(-1)),灌胃给予相应药物,假手术组、模型组给予等量的去离子水,各组均每日灌胃1次。药物连续干预8周后,采用酶联免疫吸附测定(ELISA)检测大鼠外周血中单磷酸腺苷(AMP),二磷酸腺苷(ADP),三磷酸腺苷(ATP)的含量;采用投射电镜检测心肌线粒体超微结构的变化;采用蛋白免疫印迹法(Western blot)检测大鼠心肌组织中AMPK,PPARα,PPARγ辅助激活因子1α(PGC-1α)的蛋白表达量。结果:与假手术组比较,模型组大鼠AMP,ADP含量显著升高,ATP含量显著下降(P<0.05,P<0.01);与模型组比较,补阳还五汤组、酒石酸美托洛尔组大鼠AMP含量明显降低(P<0.01),ADP含量下降(P<0.05),ATP含量升高。与假手术组比较,模型组大鼠线粒体数量减少,形态异常;与模型组比较,补阳还五汤组、酒石酸美托洛尔组大鼠线粒体数量明显增加,形态明显改善。与假手术组比较,各组大鼠AMPK蛋白表达量无统计学差异;与假手术组比较,模型组PPARα,PGC-1α蛋白表达水平明显降低(P<0.05)。与模型组比较,补阳还五汤组、酒石酸美托洛尔组PPARα,PGC-1α蛋白表达水平明显升高(P<0.05)。结论:补阳还五汤可能是通过改善线粒体结构和功能,激活AMPK并上调AMPK/PPARα信号通路的表达,从而改善衰竭心脏的能量代谢,延缓心衰进展。
Objective: To study the effect of Buyang Huanwu Tang on myocardial energy metabolism in rats with diastolic heart failure( DHF) based on adenosine monophosphate( AMP)-activated protein kinase( AMPK)/peroxisome proliferators-activated receptors α( PPARα) signaling pathway, and investigate its mechanism of action. Method: The 48 SD rats were randomly divided into sham operation group and model group.DHF rat model was established by abdominal aorta constriction method. The successfully modeled rats were randomly divided into model group,Buyang Huanwu Tang group( 12. 72 g·kg~(-1)·d~(-1)),metoprolol tartrate group( 0. 004 5 g·kg~(-1)·d~(-1)),with corresponding drugs in each group by intragastric administration. The sham operation group and model group were given with equal amount of deionized water,once a day. After 8 weeks of continuous drug intervention,the contents of adenosine monophosphate( AMP),adenosine diphosphate( ADP) and adenosine triphophate( ATP) in peripheral blood of rats were determined by enzyme linked immunosorbent assay( ELISA).The changes of myocardial mitochondrial ultrastructure were detected by electron microscope. The protein expression levels of AMPK,PPARα and peroxisome proliferator-activated receptor-γ coactivator-1α( PGC-1α) in rat myocardium were detected by Western blot. Result: As compared with sham operation group,the contents of AMP and ADP in model group were increased significantly,and ATP content was decreased significantly( P <0. 05,P < 0. 01). As compared with the model group,the AMP content was decreased significantly in Buyang Huanwu Tang group and metoprolol tartrate group( P < 0. 01),and the content of ADP was decreased while ATP content was increased( P < 0. 05). As compared with the sham operation group,the number of mitochondria in the model group was decreased,the morphology was abnormal; as compared with the model group,the number of mitochondria in Buyang Huanwu Tang group and metoprolol tartrate group was increased obviously,and the morphology was obviously improved. As compared with the sham operation group,there was no statistically significant difference in the expression of AMPK protein between various groups,but the expression levels of PPARα and PGC-1α protein in the model group were decreased significantly( P < 0. 05). As compared with the model group,the expression levels of PPARα and PGC-1α protein in Buyang Huanwu Tang group and metoprolol tartrate group were increased significantly( P < 0. 05). Conclusion: Buyang Huanwu Tang may improve the energy metabolism of the failed heart and delay the progression of heart failure by improving the structure and function of mitochondria,activating AMPK and up-regulating the expression of AMPK/PPARα signaling pathway.
Objective: To study the effect of Buyang Huanwu Tang on myocardial energy metabolism in rats with diastolic heart failure( DHF) based on adenosine monophosphate( AMP)-activated protein kinase( AMPK)/peroxisome proliferators-activated receptors α( PPARα) signaling pathway, and investigate its mechanism of action. Method: The 48 SD rats were randomly divided into sham operation group and model group.DHF rat model was established by abdominal aorta constriction method. The successfully modeled rats were randomly divided into model group,Buyang Huanwu Tang group( 12. 72 g·kg~(-1)·d~(-1)),metoprolol tartrate group( 0. 004 5 g·kg~(-1)·d~(-1)),with corresponding drugs in each group by intragastric administration. The sham operation group and model group were given with equal amount of deionized water,once a day. After 8 weeks of continuous drug intervention,the contents of adenosine monophosphate( AMP),adenosine diphosphate( ADP) and adenosine triphophate( ATP) in peripheral blood of rats were determined by enzyme linked immunosorbent assay( ELISA).The changes of myocardial mitochondrial ultrastructure were detected by electron microscope. The protein expression levels of AMPK,PPARα and peroxisome proliferator-activated receptor-γ coactivator-1α( PGC-1α) in rat myocardium were detected by Western blot. Result: As compared with sham operation group,the contents of AMP and ADP in model group were increased significantly,and ATP content was decreased significantly( P <0. 05,P < 0. 01). As compared with the model group,the AMP content was decreased significantly in Buyang Huanwu Tang group and metoprolol tartrate group( P < 0. 01),and the content of ADP was decreased while ATP content was increased( P < 0. 05). As compared with the sham operation group,the number of mitochondria in the model group was decreased,the morphology was abnormal; as compared with the model group,the number of mitochondria in Buyang Huanwu Tang group and metoprolol tartrate group was increased obviously,and the morphology was obviously improved. As compared with the sham operation group,there was no statistically significant difference in the expression of AMPK protein between various groups,but the expression levels of PPARα and PGC-1α protein in the model group were decreased significantly( P < 0. 05). As compared with the model group,the expression levels of PPARα and PGC-1α protein in Buyang Huanwu Tang group and metoprolol tartrate group were increased significantly( P < 0. 05). Conclusion: Buyang Huanwu Tang may improve the energy metabolism of the failed heart and delay the progression of heart failure by improving the structure and function of mitochondria,activating AMPK and up-regulating the expression of AMPK/PPARα signaling pathway.
引文
[1]From A M,Scott C G,CHEN H H.The development of heart failure in patients with diabetes mellitus and preclinical diastolic dysfunction:a population-based study[J].J Am Coll Cardiol,2010,55(4):300-305.
[2]Neubauer S.The failing heart—an engine out of fuel[J].New Engl J Med,2007,356(11):1140.
[3]ZHANG J,WEI C,WANG H,et al.Protective effect of qiliqiangxin capsule on energy metabolism and myocardial mitochondria in pressure overload heart failure rats[J].Evid Based Compl Alt,2013,doi:10.1155/2013/378298.
[4]Reznick R M,Shulman G I.The role of AMP-activated protein kinase in mitochondrial biogenesis[J].J Physiol,2010,574(1):33-39.
[5]平政,曹雪滨.PGC-1α活性调节的信号通路[J].国际心血管病杂志,2012,39(2):89-91.
[6]Kanda H,Nohara R,Hasegawa K,et al.A nuclear complex containing PPARalpha/RXRalpha is markedly downregulated in the hypertrophied rat left ventricular myocardium with normal systolic function[J].Heart Vessels,2000,15(4):191-196.
[7]Garnier A,Fortin D,Deloménie C,et al.Depressed mitochondrial transcription factors and oxidative capacity in rat failing cardiac and skeletal muscles[J].J Physiol,2010,551(2):491-501.
[8]Zaha V G,Young L H.AMP-activated protein kinase regulation and biological actions in the heart[J].Circ Res,2012,111(6):800-814.
[9]倪量,王硕仁,赵明镜,等.部分缩窄大鼠腹主动脉致心肌肥厚动物模型的病证结合研究[J].中国中医基础医学杂志,2007,13(10):757-759.
[10]倪量,王硕仁,赵明镜,等.腹主动脉部分缩窄大鼠模型的心肌肥厚特点[J].中国比较医学杂志,2007,17(4):214-218.
[11]李兵,柳君泽,陈丽芬.缺氧对大鼠心肌线粒体能量代谢和腺苷酸转位酶活性的影响[J].中国病理生理杂志,2006,22(3):460-463.
[12]张军芳,王磊,魏聪,等.慢性心力衰竭能量代谢重构与治疗进展[J].中国老年学杂志,2014,34(4):1115-1117.
[13]Reznick R M,Shulman G I.The role of AMP-activated protein kinase inmitochondrial biogenesis[J].J Physiol,2010,574(1):33-39.
[14]Alp P R,Newsholme E A,Zammit V A.Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates[J].Biochem J,1976,154(3):689-700.
[15]Rowe G C,Jiang A,Arany Z.PGC-1 coactivators in cardiac development and disease[J].Circ Res,2010,107(7):825-838.
[16]张明丽,姜瑞雪.益气活血汤对慢性心力衰竭气虚血瘀证患者心功能及生活质量的影响[J].中国实验方剂学杂志,2015,21(19):162-166.
[17]王懿,张艳,礼海.益气活血方干预PGC-1α调控心衰心肌细胞能量代谢重构的作用机制[J].中国实验方剂学杂志,2015,21(6):169-173.
[2]Neubauer S.The failing heart—an engine out of fuel[J].New Engl J Med,2007,356(11):1140.
[3]ZHANG J,WEI C,WANG H,et al.Protective effect of qiliqiangxin capsule on energy metabolism and myocardial mitochondria in pressure overload heart failure rats[J].Evid Based Compl Alt,2013,doi:10.1155/2013/378298.
[4]Reznick R M,Shulman G I.The role of AMP-activated protein kinase in mitochondrial biogenesis[J].J Physiol,2010,574(1):33-39.
[5]平政,曹雪滨.PGC-1α活性调节的信号通路[J].国际心血管病杂志,2012,39(2):89-91.
[6]Kanda H,Nohara R,Hasegawa K,et al.A nuclear complex containing PPARalpha/RXRalpha is markedly downregulated in the hypertrophied rat left ventricular myocardium with normal systolic function[J].Heart Vessels,2000,15(4):191-196.
[7]Garnier A,Fortin D,Deloménie C,et al.Depressed mitochondrial transcription factors and oxidative capacity in rat failing cardiac and skeletal muscles[J].J Physiol,2010,551(2):491-501.
[8]Zaha V G,Young L H.AMP-activated protein kinase regulation and biological actions in the heart[J].Circ Res,2012,111(6):800-814.
[9]倪量,王硕仁,赵明镜,等.部分缩窄大鼠腹主动脉致心肌肥厚动物模型的病证结合研究[J].中国中医基础医学杂志,2007,13(10):757-759.
[10]倪量,王硕仁,赵明镜,等.腹主动脉部分缩窄大鼠模型的心肌肥厚特点[J].中国比较医学杂志,2007,17(4):214-218.
[11]李兵,柳君泽,陈丽芬.缺氧对大鼠心肌线粒体能量代谢和腺苷酸转位酶活性的影响[J].中国病理生理杂志,2006,22(3):460-463.
[12]张军芳,王磊,魏聪,等.慢性心力衰竭能量代谢重构与治疗进展[J].中国老年学杂志,2014,34(4):1115-1117.
[13]Reznick R M,Shulman G I.The role of AMP-activated protein kinase inmitochondrial biogenesis[J].J Physiol,2010,574(1):33-39.
[14]Alp P R,Newsholme E A,Zammit V A.Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates[J].Biochem J,1976,154(3):689-700.
[15]Rowe G C,Jiang A,Arany Z.PGC-1 coactivators in cardiac development and disease[J].Circ Res,2010,107(7):825-838.
[16]张明丽,姜瑞雪.益气活血汤对慢性心力衰竭气虚血瘀证患者心功能及生活质量的影响[J].中国实验方剂学杂志,2015,21(19):162-166.
[17]王懿,张艳,礼海.益气活血方干预PGC-1α调控心衰心肌细胞能量代谢重构的作用机制[J].中国实验方剂学杂志,2015,21(6):169-173.