基于PI3K/Akt/mTOR信号通路研究南葶苈子水提液对心力衰竭模型大鼠心室重构的影响
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摘要
目的观察南葶苈子水提液对压力后负荷性心衰模型大鼠心室重构的影响,探讨PI3K/Akt/mTOR信号通路在其中的作用。方法雄性SD大鼠15只,按随机数字表法分为正常组、模型组和南葶苈子组,每组5只。SD大鼠心力衰竭动物模型建立后,正常组及模型组大鼠给予5mL·kg~(-1)·d~(-1)生理盐水灌胃,葶苈子组大鼠给予南葶苈子水提液10g·kg~(-1)·d~(-1)灌胃,共4周。4周后处死大鼠,测定各组大鼠心室肥厚指数;HE染色法和电镜观察心室组织病理变化;Western blot检测各组大鼠左心室心肌组织Bax、Bcl-2、Cleaved-caspase 3、p-Akt、Akt、p-mTOR、mTOR蛋白表达水平。结果南葶苈子组大鼠心室肥厚指数较模型组降低[(0.0033±0.0002)比(0.0037±0.0001),P<0.05];HE染色和电镜显示,南葶苈子组心肌纤维排列不规则和心肌细胞肥厚程度较模型组减轻;Western blot证实,南葶苈子组大鼠Bcl-2/Bax蛋白表达灰度值较模型组上调[(0.75±0.20)比(0.54±0.18),P<0.05];Caspase-3/GAPDH蛋白表达灰度值较模型组下调[(1.31±0.22)比(1.62±0.26),P<0.05];p-Akt/Akt和p-mTOR/mTOR蛋白表达灰度值较模型组上调[p-Akt/Akt:(1.50±0.18)比(1.18±0.21),P<0.05;p-mTOR/mTOR:(1.36±0.32)比(1.16±0.12),P<0.05]。结论南葶苈子水提液能改善压力后负荷性心衰模型大鼠心室重构,其作用机制可能与激活PI3K/Akt/mTOR信号通路抑制心肌细胞凋亡有关。
Objective To observe the effect of aqueous extract from seeds of Descurainia sophia on ventricular remodeling in rats with pressure afterload heart failure and explore the role of PI3K/Akt/mTOR signaling pathway.Methods Fifteen male Sprague-Dawley(SD)rats were divided into normal group,model group and Descurainia sophia group according to random number table method,each had 5 individuals.After the establishment of SD rat model of heart failure,rats in the normal group and model group were given 5ml·kg~(-1)·d~(-1)normal saline gavage administration for 4 weeks,and the Descurainia sophia group was given aqueous extract from seeds of Descurainia sophia instead.Four weeks later,rats were sacrificed and tested.The ventricular hypertrophy index of rats in each group was determined,the pathological changes of ventricular tissue were observed by HE staining and electron microscopy,and the expression levels of Bax,Bcl-2,Cleaved-caspase 3,p-Akt,Akt,p-mTOR and mTOR in left ventricular myocardium were detected by Western blot.ResultsThe ventricular hypertrophy index of rats in the Descurainia sophia group was lower than that in the model group,(0.0033±0.0002)vs(0.0037±0.0001),P<0.05.HE staining and electron microscopy showed that the irregular arrangement of myocardial fibers and the degree of hypertrophy of myocardial cells in the group of Amaranth were less than those in the model group.Western blot revealed that the gray value of Bcl-2/Bax protein expression in the Descurainia sophia group was higher than that in the model group,(0.75±0.20)vs(0.54±0.18),P<0.05;the gray valu of caspase-3/GAPDH expression was lower,(1.31±0.22)vs(1.62±0.26),P<0.05;the gray values of p-Akt/Akt and p-mTOR/mTOR expression were higher,(1.50±0.18)vs(1.18±0.21),(1.36±0.32)vs(1.16±0.12),P<0.05,respectively.Conclusion Aqueous extract from seeds of Descurainia sophia can improve ventricular remodeling in rats with pressure-overload heart failure.Its mechanism may be related to the inhibition of cardiomyocyte apoptosis by activating PI3K/Akt/mTOR signaling pathway.
Objective To observe the effect of aqueous extract from seeds of Descurainia sophia on ventricular remodeling in rats with pressure afterload heart failure and explore the role of PI3K/Akt/mTOR signaling pathway.Methods Fifteen male Sprague-Dawley(SD)rats were divided into normal group,model group and Descurainia sophia group according to random number table method,each had 5 individuals.After the establishment of SD rat model of heart failure,rats in the normal group and model group were given 5ml·kg~(-1)·d~(-1)normal saline gavage administration for 4 weeks,and the Descurainia sophia group was given aqueous extract from seeds of Descurainia sophia instead.Four weeks later,rats were sacrificed and tested.The ventricular hypertrophy index of rats in each group was determined,the pathological changes of ventricular tissue were observed by HE staining and electron microscopy,and the expression levels of Bax,Bcl-2,Cleaved-caspase 3,p-Akt,Akt,p-mTOR and mTOR in left ventricular myocardium were detected by Western blot.ResultsThe ventricular hypertrophy index of rats in the Descurainia sophia group was lower than that in the model group,(0.0033±0.0002)vs(0.0037±0.0001),P<0.05.HE staining and electron microscopy showed that the irregular arrangement of myocardial fibers and the degree of hypertrophy of myocardial cells in the group of Amaranth were less than those in the model group.Western blot revealed that the gray value of Bcl-2/Bax protein expression in the Descurainia sophia group was higher than that in the model group,(0.75±0.20)vs(0.54±0.18),P<0.05;the gray valu of caspase-3/GAPDH expression was lower,(1.31±0.22)vs(1.62±0.26),P<0.05;the gray values of p-Akt/Akt and p-mTOR/mTOR expression were higher,(1.50±0.18)vs(1.18±0.21),(1.36±0.32)vs(1.16±0.12),P<0.05,respectively.Conclusion Aqueous extract from seeds of Descurainia sophia can improve ventricular remodeling in rats with pressure-overload heart failure.Its mechanism may be related to the inhibition of cardiomyocyte apoptosis by activating PI3K/Akt/mTOR signaling pathway.
引文
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[9]Wang Y,Hu Y,Zeng Z,et al.Influence of androgen on myocardial apoptosis and expression of myocardial IR and IRS1 in chronic heart failure rat models[J].Mol Med Rep,2018,17(1):1057-1064.
[10]Ito T,Schaffer S,Azuma J.The effect of taurine on chronic heart failure:actions of taurine against catecholamine and angiotensin II[J].Amino Acids,2013,46(1):111-119.
[11]Maccormack TJ,Callaghan NI,Sykes AV,et al.Taurine depresses cardiac contractility and enhances systemic heart glucose utilization in the cuttlefish,Sepia officinalis[J].Journal of Comparative Physiology B,2016,186(2):215-227.
[12]Green DR,Fitzgerald P.Just So Stories about the Evolution of Apoptosis[J].Curr Biol,2016,26(13):R620-R627.
[13]Lee HJ,Lee EK,Seo YE,et al.Roles of Bcl-2 and caspase-9 and-3 in CD30-induced human eosinophil apoptosis[J].J Microbiol Immunol Infect,2017,50(2):145-152.
[14]Asati V,Mahapatra DK,Bharti SK.PI3K/Akt/mTOR and Ras/Raf/MEK/ERK signaling pathways inhibitors as anticancer agents:Structural and pharmacological perspectives[J].Eur J Med Chem,2016,109:314-341.
[2]Zhou N,Sun YP,Zheng XK,et al.A Metabolomics-Based Strategy for the Mechanism Exploration of Traditional Chinese Medicine:Descurainia sophia Seeds Extract and Fractions as a Case Study[J].Evidence-Based Complementray and Alternative Medicine,2017,2017:2845173.
[3]Ponikowski P,Anker SD,Bueno H,et al.2016 ESC Guidelines for the diagnosis and treatment of acute and chronic heart failure[J].European Heart Journal,2016,37(27):2129-2200.
[4]郭娟,陈长勋.葶苈子对压力负荷性大鼠心室重构及神经内分泌因子和心肌Ⅰ、Ⅲ型胶原的影响[J].中药材,2007,30(8):963-967.
[5]郭娟,陈长勋,沈云辉.葶苈子水提液对动物实验性心室重构的影响[J].中草药,2007,38(10):1519-1523.
[6]孙志强,李超,吴源鸿,等.南葶苈子提取液对心力衰竭大鼠心功能的影响[J].心脑血管病防治,2016,16(6):427-430.
[7]Lin YY,Cheng YJ,Hu J,et al.The Coexistence of Hypertension and Ovariectomy Additively Increases Cardiac Apoptosis[J].Int J Mol Sci,2016,17(12):E2036.
[8]Gupte M,Lal H,Ahmad F,et al.Chronic Neuregulin-1βTreatment Mitigates the Progression of Postmyocardial Infarction Heart Failure in the Setting of Type 1 Diabetes Mellitus by Suppressing Myocardial Apoptosis,Fibrosis,and Key Oxidant-Producing Enzymes[J].J Card Fail,2017,23(12):887-899.
[9]Wang Y,Hu Y,Zeng Z,et al.Influence of androgen on myocardial apoptosis and expression of myocardial IR and IRS1 in chronic heart failure rat models[J].Mol Med Rep,2018,17(1):1057-1064.
[10]Ito T,Schaffer S,Azuma J.The effect of taurine on chronic heart failure:actions of taurine against catecholamine and angiotensin II[J].Amino Acids,2013,46(1):111-119.
[11]Maccormack TJ,Callaghan NI,Sykes AV,et al.Taurine depresses cardiac contractility and enhances systemic heart glucose utilization in the cuttlefish,Sepia officinalis[J].Journal of Comparative Physiology B,2016,186(2):215-227.
[12]Green DR,Fitzgerald P.Just So Stories about the Evolution of Apoptosis[J].Curr Biol,2016,26(13):R620-R627.
[13]Lee HJ,Lee EK,Seo YE,et al.Roles of Bcl-2 and caspase-9 and-3 in CD30-induced human eosinophil apoptosis[J].J Microbiol Immunol Infect,2017,50(2):145-152.
[14]Asati V,Mahapatra DK,Bharti SK.PI3K/Akt/mTOR and Ras/Raf/MEK/ERK signaling pathways inhibitors as anticancer agents:Structural and pharmacological perspectives[J].Eur J Med Chem,2016,109:314-341.