黄芪注射液调节PI3K/AKT通路改善心肌梗死后心肌重塑
|
摘要
目的探讨黄芪注射液(Astragalus injection,AS)对大鼠心肌梗死后心肌重塑的逆转作用及其作用机制。方法通过结扎左冠状动脉前降支建立大鼠心肌梗死后心肌重塑模型,实验分为假手术组、模型组、AS低剂量组(1 m L/kg)及高剂量组(3 m L/kg)。研究AS治疗6周对其生存百分率、血清中α肌球蛋白重链(α-MHC)和大鼠N端前脑钠素(NT-PROBNP)的影响;同时HE染色及Masson染色以检测心脏组织病理学变化及其心肌纤维化情况; ELISA法检测各组血清中MDA、NO及e NOS的变化; Western blotting法检测心肌细胞PI3K、P-AKT、及Pe NOS蛋白的表达。结果 AS高低剂量组与模型组比较,生存百分率有所提高,但无显著性差异;而AS高剂量组α-MHC及NT-PROBNP较模型组显著降低(P<0. 05); HE及Masson染色形态学检查显示,与模型组比较,AS高剂量组炎细胞浸润、出血及纤维化程度减轻;高剂量组血清中MDA降低(P<0. 001)、NO及e NOS的升高(P<0. 05); Western blot结果显示,AS组的PI3K、p-AKT表达量高于模型组(P<0. 05),p-eNOS表达亦显著上升。结论黄芪注射液可能通过激活PI3K/AKT通路降低氧化应激水平,从而抑制心肌梗死后心肌重塑。
Objective To examine the effect of Astragalus( AS) injection on post-myocardial infarction-induced cardiac remodeling in rats,and explore potential mechanisms underlying such effects. Methods A cardiac remodeling post-myocardial infarction model was prepared in rats,which were divided into four groups: sham operation,myocardial infarction( model group),low-dose AS( ASL,1 m L/kg),and high-dose AS( ASH,3 m L/kg). Survival rate of the rats and activities of the alpha isoform of myosin heavy chain( α-MHC) and N-terminal pro b-type natriuretic peptide( NTpro BNP) in rat serum were measured after 6-week treatment. Histopathological changes and myocardial fibrosis were observed by light microscopy using hematoxylin and eosin,and Masson trichome staining. Quantification of malondialdehyde( MDA),nitric oxide( NO),and endothelial nitric oxide synthase( e NOS) were carried out using enzyme-linked immunosorbent assays. Protein expression of phosphatidylinositol 3 kinase( PI3 K),phosphorylated AKT( p-AKT),and pe NOS were analyzed by western blotting. Results Compared with the model group,the survival rate of the ASL and ASH group were increased,but not significantly( P > 0. 05). Meanwhile,α-MHC and NT-proBNP contents in the ASH group were reduced( P < 0. 05). In addition,inflammation,hyperemia,and interstitial fibrosis were reduced in the ASH group.Moreover,the ASH group exhibited increased protein expression of PI3 K and p-AKT in the injured heart( P < 0. 05). pe NOS expression in the ASH group was increased compared with the model group( P < 0. 05). Conclusions AS elicits an obvious protective effect on cardiac remodeling after myocardial infarction in rats. Activation of the PI3 K/AKT pathway may activate p-eNOS,which is involved in the mechanism by which AS prevents oxidative stress during cardiac remodeling.
Objective To examine the effect of Astragalus( AS) injection on post-myocardial infarction-induced cardiac remodeling in rats,and explore potential mechanisms underlying such effects. Methods A cardiac remodeling post-myocardial infarction model was prepared in rats,which were divided into four groups: sham operation,myocardial infarction( model group),low-dose AS( ASL,1 m L/kg),and high-dose AS( ASH,3 m L/kg). Survival rate of the rats and activities of the alpha isoform of myosin heavy chain( α-MHC) and N-terminal pro b-type natriuretic peptide( NTpro BNP) in rat serum were measured after 6-week treatment. Histopathological changes and myocardial fibrosis were observed by light microscopy using hematoxylin and eosin,and Masson trichome staining. Quantification of malondialdehyde( MDA),nitric oxide( NO),and endothelial nitric oxide synthase( e NOS) were carried out using enzyme-linked immunosorbent assays. Protein expression of phosphatidylinositol 3 kinase( PI3 K),phosphorylated AKT( p-AKT),and pe NOS were analyzed by western blotting. Results Compared with the model group,the survival rate of the ASL and ASH group were increased,but not significantly( P > 0. 05). Meanwhile,α-MHC and NT-proBNP contents in the ASH group were reduced( P < 0. 05). In addition,inflammation,hyperemia,and interstitial fibrosis were reduced in the ASH group.Moreover,the ASH group exhibited increased protein expression of PI3 K and p-AKT in the injured heart( P < 0. 05). pe NOS expression in the ASH group was increased compared with the model group( P < 0. 05). Conclusions AS elicits an obvious protective effect on cardiac remodeling after myocardial infarction in rats. Activation of the PI3 K/AKT pathway may activate p-eNOS,which is involved in the mechanism by which AS prevents oxidative stress during cardiac remodeling.
引文
[1] Palazzuoli A,Nuti R. Heart failure:pathophysiology and clinical picture[J]. Contrib Nephrol,2010; 164:1-10.
[2]王萧,林传权,许庆文,等. Caspase-3在心衰兔模型细胞凋亡过程中的变化[J].中国实验动物学报,2009;(3):201-205
[3] Cao DJ,Wang ZV,Battiproluetal PK,et al. Histone deacetylase(HDAC)inhibitors attenuate cardiac hypertrophy by suppressing autophagy[J]. Proc Natl Acad Sci U S A,2011,108(10):4123-4128.
[4]顾文超,周晓慧,林芳,等. T细胞亚群在心室重构中的作用研究进展[J].中国比较医学杂志,2017,(10):85-88.
[5] Hori M, Nishida K. Oxidative stress and left ventricular remodelling after myocardial infarction[J]. Cardiovasc Res,2009,81(3):457-464.
[6] Anilkumar N, Sirker A, Shah AM,et al. Redox sensitive signaling pathways in cardiac remodeling,hypertrophy and failure[J]. Front Biosci(Landmark Ed),2009,14:3168-3187.
[7] Seddon M,Looi YH,Shah AM. Oxidative stress and redox signalling in cardiac hypertrophy and heart failure[J]. Heart,2007 93(8):903-907.
[8] Zhang Y,Tocchetti CG,Krieg T,et al. Oxidative and nitrosative stress in the maintenance of myocardial function[J]. Free Radical Biol Med,2012,53(8); 1531-1540.
[9]原广平,黄平.黄芪注射液治疗慢性心力衰竭的临床疗效评价[J].中国临床药理学与治疗学,2003,8(6):710-711.
[10]曾和松,吴正贤,刘正湘,等.黄芪注射液治疗心功能不全并心律失常的临床观察[J].实用心脑肺血管病杂志,2003,11,(2):71-72.
[11]关凤英,李红,于秀霞,等.黄芪注射液对心肌细胞氧化应激性损伤的保护作用[J].中国康复理论与实践,2010,16(9):830-832.
[12] Jia Y,Zuo D,Li Z,et al. Astragaloside IV inhibits doxorubicininduced cardiomyocyte apoptosis mediated by mitochondrial apoptotic pathway via activating the PI3K/Akt pathway[J].Chem Pharm Bull,2014,62(1):45-53.
[13] Park CW,Kim HW,Lim JH,et al. Vascular endothelial growth factor inhibition by dRK6 causes endothelial apoptosis,fibrosis,and inflammation in the heart via the Akt/e NOS axis in db/db mice[J]. Diabetes,2009,58(11):2666-2676.
[14]郑海娟,王维亭,郝春华,等.地奥心血康对大鼠心肌缺血再灌注损伤的治疗作用[J].天津医药,2015, 43(5):491-495.
[15] Mozaffari MS,Liu JY,Schaffer SW. Effect of pressure overload on cardioprotection via PI3K-Akt:comparison of postconditioning,insulin,and pressure unloading[J]. Am J Hypertens,2010,23(6):668-674.
[16]赵婧,屈艺,母得志. Akt与细胞线粒体凋亡途径[J].生命的化学,2010,30(4):528-532.
[17] Datta SR,Dudek H,Tao X,et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery[J]. Cell,1997,91(2):231-241.
[18] Almhanna K,Strosberg J,Malafa M. Targeting AKT protein kinase in gastric cancer[J]. Anticancer Res,2011,31(12):4387-4392.
[19] Mount PF,Kemp BE,Power DA. Regulation of endothelial and myocardial NO synthesis by multi-site e NOS phosphorylation[J]. J Mol Cell Cardiol,2007,42(2):271-279.
[20] Zhu M,Feng J,Lucchinetti E,et al. Ischemic postconditioning protects remodeled myocardium via the PI3KPKB/Akt reperfusion injury salvage kinase pathway[J]. Cardiovasc Res,2006; 72(1):152-162.
[21] Zancheta D, Troiano JA, Potje SR, The PI3K-Akt-eNOS pathway is involved in aortic hyporeactivity to phenylephrine associated with late pregnancy in spontaneously hypertensive rats[J]. Life Sci,2015,122:78-86.
[22] Ahsan A, Han G, Pan J, et al. Phosphocreatine protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/e NOS pathway[J].Apoptosis,2015,20(12):1563-1576.
[23] Otani H. The role of nitric oxide in myocardial repair and remodeling[J]. Antioxid Redox Signal,2009,11(8):1913-1928.
[24] Chen LL,Zhu TB,Yin H,et,al. Inhibition of MAPK signaling by e NOS gene transfer improves ventricular remodeling after myocardial infarction through reduction of inflammation[J]. Mol Biol Rep,2010,37(7):3067-3072.
[25] Takimoto E,Champion HC,Li M,et al. Oxidant stress from nitric oxide synthase-3 uncoupling stimulates cardiac pathologic remodeling from chronic pressure load[J]. J Clin Invest,2005,115(5):1221-1231.
[26] Nediani C,Raimondi L,Borchi E,et al. Nitric oxide/reactive oxygen species generation and nitroso/redox imbalance in heart failure:from molecular mechanisms to therapeutic implications[J]. Antioxid Redox Signal,2011,14(2):289-331.
[2]王萧,林传权,许庆文,等. Caspase-3在心衰兔模型细胞凋亡过程中的变化[J].中国实验动物学报,2009;(3):201-205
[3] Cao DJ,Wang ZV,Battiproluetal PK,et al. Histone deacetylase(HDAC)inhibitors attenuate cardiac hypertrophy by suppressing autophagy[J]. Proc Natl Acad Sci U S A,2011,108(10):4123-4128.
[4]顾文超,周晓慧,林芳,等. T细胞亚群在心室重构中的作用研究进展[J].中国比较医学杂志,2017,(10):85-88.
[5] Hori M, Nishida K. Oxidative stress and left ventricular remodelling after myocardial infarction[J]. Cardiovasc Res,2009,81(3):457-464.
[6] Anilkumar N, Sirker A, Shah AM,et al. Redox sensitive signaling pathways in cardiac remodeling,hypertrophy and failure[J]. Front Biosci(Landmark Ed),2009,14:3168-3187.
[7] Seddon M,Looi YH,Shah AM. Oxidative stress and redox signalling in cardiac hypertrophy and heart failure[J]. Heart,2007 93(8):903-907.
[8] Zhang Y,Tocchetti CG,Krieg T,et al. Oxidative and nitrosative stress in the maintenance of myocardial function[J]. Free Radical Biol Med,2012,53(8); 1531-1540.
[9]原广平,黄平.黄芪注射液治疗慢性心力衰竭的临床疗效评价[J].中国临床药理学与治疗学,2003,8(6):710-711.
[10]曾和松,吴正贤,刘正湘,等.黄芪注射液治疗心功能不全并心律失常的临床观察[J].实用心脑肺血管病杂志,2003,11,(2):71-72.
[11]关凤英,李红,于秀霞,等.黄芪注射液对心肌细胞氧化应激性损伤的保护作用[J].中国康复理论与实践,2010,16(9):830-832.
[12] Jia Y,Zuo D,Li Z,et al. Astragaloside IV inhibits doxorubicininduced cardiomyocyte apoptosis mediated by mitochondrial apoptotic pathway via activating the PI3K/Akt pathway[J].Chem Pharm Bull,2014,62(1):45-53.
[13] Park CW,Kim HW,Lim JH,et al. Vascular endothelial growth factor inhibition by dRK6 causes endothelial apoptosis,fibrosis,and inflammation in the heart via the Akt/e NOS axis in db/db mice[J]. Diabetes,2009,58(11):2666-2676.
[14]郑海娟,王维亭,郝春华,等.地奥心血康对大鼠心肌缺血再灌注损伤的治疗作用[J].天津医药,2015, 43(5):491-495.
[15] Mozaffari MS,Liu JY,Schaffer SW. Effect of pressure overload on cardioprotection via PI3K-Akt:comparison of postconditioning,insulin,and pressure unloading[J]. Am J Hypertens,2010,23(6):668-674.
[16]赵婧,屈艺,母得志. Akt与细胞线粒体凋亡途径[J].生命的化学,2010,30(4):528-532.
[17] Datta SR,Dudek H,Tao X,et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery[J]. Cell,1997,91(2):231-241.
[18] Almhanna K,Strosberg J,Malafa M. Targeting AKT protein kinase in gastric cancer[J]. Anticancer Res,2011,31(12):4387-4392.
[19] Mount PF,Kemp BE,Power DA. Regulation of endothelial and myocardial NO synthesis by multi-site e NOS phosphorylation[J]. J Mol Cell Cardiol,2007,42(2):271-279.
[20] Zhu M,Feng J,Lucchinetti E,et al. Ischemic postconditioning protects remodeled myocardium via the PI3KPKB/Akt reperfusion injury salvage kinase pathway[J]. Cardiovasc Res,2006; 72(1):152-162.
[21] Zancheta D, Troiano JA, Potje SR, The PI3K-Akt-eNOS pathway is involved in aortic hyporeactivity to phenylephrine associated with late pregnancy in spontaneously hypertensive rats[J]. Life Sci,2015,122:78-86.
[22] Ahsan A, Han G, Pan J, et al. Phosphocreatine protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/e NOS pathway[J].Apoptosis,2015,20(12):1563-1576.
[23] Otani H. The role of nitric oxide in myocardial repair and remodeling[J]. Antioxid Redox Signal,2009,11(8):1913-1928.
[24] Chen LL,Zhu TB,Yin H,et,al. Inhibition of MAPK signaling by e NOS gene transfer improves ventricular remodeling after myocardial infarction through reduction of inflammation[J]. Mol Biol Rep,2010,37(7):3067-3072.
[25] Takimoto E,Champion HC,Li M,et al. Oxidant stress from nitric oxide synthase-3 uncoupling stimulates cardiac pathologic remodeling from chronic pressure load[J]. J Clin Invest,2005,115(5):1221-1231.
[26] Nediani C,Raimondi L,Borchi E,et al. Nitric oxide/reactive oxygen species generation and nitroso/redox imbalance in heart failure:from molecular mechanisms to therapeutic implications[J]. Antioxid Redox Signal,2011,14(2):289-331.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |