丹参酮ⅡA对扩张型心肌病大鼠心肌细胞凋亡及PI3K/Akt通路的影响
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摘要
目的探究丹参酮ⅡA(STS)对扩张型心肌病(DCM)大鼠心肌细胞凋亡的改善作用以及可能的作用机制。方法制备DCM大鼠,随机数字表法分为DCM组、STS-L、STS-M、STS-H、卡维地洛组(CAR),另设置对照组,每组均12只大鼠,超声检测左心室收缩末期内径(LVIDs)和左心室舒张末期内径(LVIDd)、左心室射血分数(LVEF),HE、Masson染色观察心肌组织病理损伤情况; ELISA法检测血清中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平; TUNEL染色法检测大鼠心肌细胞凋亡情况;免疫印迹(WB)、免疫组化法检测心肌组织中p-磷脂酰肌醇3激酶(PI3K)、p-蛋白激酶B(Akt)、Bax、caspase-3、Bcl-2表达。结果与对照组相比,DCM组LVIDs、LVIDd增大,LVEF降低,心肌组织病理学评分、心肌胶原纤维比例均升高,血清TNF-ɑ、IL-6水平升高,心肌细胞AI升高(P<0. 05);与DCM组相比,STS各组以及CAR组LVIDs、LVIDd降低,LVEF提高,心肌组织病理学评分、心肌胶原纤维比例均降低,血清TNF-ɑ、IL-6水平降低,心肌细胞AI降低(P<0. 05)。与对照组相比,DCM组p-PI3K、p-Akt、Bcl-2蛋白表达降低,caspase3、Bax蛋白表达升高(P<0. 05);与DCM组相比,STS各组以及CAR组p-PI3K、p-Akt、Bcl-2蛋白表达升高,caspase3、Bax蛋白表达降低(P<0. 05)。结论 STS可能通过激活PI3K/Akt信号通路,抑制DCM大鼠心肌细胞凋亡,发挥对DCM大鼠的保护作用。
Objective To investigate the effect of sodium tanshinone Ⅱ A sulfonate( STS) on cardiomyocyte apoptosis in rats with dilated cardiomyopathy( DCM),and its potential mechanism of action. Methods DCM rats were randomly divided into DCM,STS-L,STS-M,STS-H,Carvedilol( CAR),and control groups,12 rats in each group. Left ventricular internal diameter at end-systole( LVIDs) and left ventricular internal dimension at end-diastole( LVIDd),and left ventricular ejection fraction( LVEF) were measured by echocardiography. Pathological injury of myocardial tissues was observed using hematoxylin and eosin,and Masson trichrome staining. Tumor necrosis factor-α( TNF-α) and interleukin-6( IL-6) levels were quantified using enzyme-linked immunosorbent assays,while apoptosis in cardiomyocytes was detected using TUNEL staining. Expressions of phosphorylated phosphatidylinositol 3 kinase( p-PI3 K),phosphorylated protein kinase B( p-Akt),Bax,caspase-3,and Bcl-2 were detected by immunoblotting and immunohistochemistry. Results Compared with the control group,the DCM group exhibited increased LVIDs and LVIDd,decreased LVEF,increased myocardial histopathologic scores and percentage of cardiac collagen fibers,increased serum levels of TNF-ɑ and IL-6,and increased apoptosis index( AI) in cardiomyocytes( P < 0. 05). Compared with the DCM group,the STS and CAR groups exhibited decreased LVIDs and LVIDd,increased LVEF,decreased myocardial histopathologic scores and percentage of cardiac collagen fibers,decreased serum levels of TNF-ɑ and IL-6,and decreased apoptosis index( AI) in cardiomyocytes( P < 0. 05). Compared with the control group,the expressions of p-PI3 K,p-Akt,and Bcl-2 proteins were decreased in the DCM group,while expressions of caspase-3 and Bax proteins were increased( P < 0. 05). Compared with the DCM group,the expressions of p-PI3 K,p-Akt,and Bcl-2 proteins in STS and CAR groups were increased,while expressions of caspase 3 and Bax proteins were decreased( P < 0. 05). Conclusions STS may inhibit cardiomyocyte apoptosis in DCM rats by activating PI3 K/Akt signaling and exert protective effects.
Objective To investigate the effect of sodium tanshinone Ⅱ A sulfonate( STS) on cardiomyocyte apoptosis in rats with dilated cardiomyopathy( DCM),and its potential mechanism of action. Methods DCM rats were randomly divided into DCM,STS-L,STS-M,STS-H,Carvedilol( CAR),and control groups,12 rats in each group. Left ventricular internal diameter at end-systole( LVIDs) and left ventricular internal dimension at end-diastole( LVIDd),and left ventricular ejection fraction( LVEF) were measured by echocardiography. Pathological injury of myocardial tissues was observed using hematoxylin and eosin,and Masson trichrome staining. Tumor necrosis factor-α( TNF-α) and interleukin-6( IL-6) levels were quantified using enzyme-linked immunosorbent assays,while apoptosis in cardiomyocytes was detected using TUNEL staining. Expressions of phosphorylated phosphatidylinositol 3 kinase( p-PI3 K),phosphorylated protein kinase B( p-Akt),Bax,caspase-3,and Bcl-2 were detected by immunoblotting and immunohistochemistry. Results Compared with the control group,the DCM group exhibited increased LVIDs and LVIDd,decreased LVEF,increased myocardial histopathologic scores and percentage of cardiac collagen fibers,increased serum levels of TNF-ɑ and IL-6,and increased apoptosis index( AI) in cardiomyocytes( P < 0. 05). Compared with the DCM group,the STS and CAR groups exhibited decreased LVIDs and LVIDd,increased LVEF,decreased myocardial histopathologic scores and percentage of cardiac collagen fibers,decreased serum levels of TNF-ɑ and IL-6,and decreased apoptosis index( AI) in cardiomyocytes( P < 0. 05). Compared with the control group,the expressions of p-PI3 K,p-Akt,and Bcl-2 proteins were decreased in the DCM group,while expressions of caspase-3 and Bax proteins were increased( P < 0. 05). Compared with the DCM group,the expressions of p-PI3 K,p-Akt,and Bcl-2 proteins in STS and CAR groups were increased,while expressions of caspase 3 and Bax proteins were decreased( P < 0. 05). Conclusions STS may inhibit cardiomyocyte apoptosis in DCM rats by activating PI3 K/Akt signaling and exert protective effects.
引文
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[8]刘崇斌,王伊林,柴花,等.丹参多酚酸盐对阿霉素诱导的扩张型心肌病大鼠心功能的影响及机制研究[J].临床心血管病杂志,2016,32(2):134-138.
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[13]曹慧敏,宋囡,张妮,等.丹参酮Ⅱ_A通过PI3K/Akt/m TOR信号通路调控自噬抗内皮细胞氧化应激损伤研究[J].北京中医药大学学报,2017,40(11):933-939.
[14]金云晔,翟昌林,沈震,等.丹参酮ⅡA磺酸钠对大鼠缺血再灌注心肌细胞凋亡的影响[J].中华中医药学刊,2014,32(6):1414-1416.
[15]李佳. PI3K/AKT/GSK3-b信号转导通路在红景天苷保护大鼠心肌缺血再灌注损伤中的作用机制[J].中西医结合心脑血管病杂志,2016,14(22):2621-2624.
[16]韩军,宣佳利,胡浩然,等.金丝桃苷预处理减轻大鼠心肌缺血再灌注损伤作用与PI3K/Akt信号通路的关系[J].中国中药杂志,2015,40(1):118-123.
[17] Opferman JT. Attacking cancer’s Achilles heel:Antagonism of anti-apoptotic BCL-2 family members[J]. FEBS J,2016,283(14):2661-2675.
[18] Vasilikos L,Spilgies LM,Knop J,et al. Regulating the balance between necroptosis,apoptosis and inflammation by inhibitors of apoptosis proteins[J]. Immunol Cell Biol,2017,95(2):160-165.
[19]王厚磊,卢伟,李德芳,等.大鼠椎间盘髓核细胞体外凋亡模型的建立[J].中国实验动物学报,2015,23(6):607-611.
[2] Metzger M,Higuchi ML, Moreira LF, et al. Relevance of apoptosis and cell proliferation for survival of patients with dilated cardiomyopathy undergoing partial left ventriculectomy[J]. Eur J Clin Invest,2002,32(6):394-399.
[3]段媛媛,郭振丰,李雪连.丹参酮ⅡA治疗心血管疾病研究机制新进展[J].中国临床药理学杂志,2016,32(19):1817-1820.
[4]黄荣杰,刘唐威,伍伟锋,等.吠喃哇酮制备大鼠扩张型心肌病模型[J].中国病理生理杂志,2003,23(10):2147-2150.
[5]杨杰孚,廖玉华,袁璟,等.中国扩张型心肌病诊断和治疗指南[J].临床心血管病杂志,2018,34(5):421-434.
[6] Arumugam S, Sreedhar R, Karuppagounder V, et al.Comparative evaluation of torasemide and spironolactone on adverse cardiac remodeling in a rat model of dilated cardiomyopathy[J]. Cardiovasc Ther,2017,35(5):12283.
[7] Liu Z,Wang J,Huang E,et al. Tanshinone IIA suppresses cholesterol accumulation in human macrophages:role of heme oxygenase-1[J]. J Lipid Res,2014,55(2):201-213..
[8]刘崇斌,王伊林,柴花,等.丹参多酚酸盐对阿霉素诱导的扩张型心肌病大鼠心功能的影响及机制研究[J].临床心血管病杂志,2016,32(2):134-138.
[9] Gill SE,Rohan M,Mehta S. Role of pulmonary microvascular endothelial cell apoptosis in murine sepsis-induced lung injury in vivo[J]. Respir Res,2015,16(1):109.
[10] Hassan MQ,Akhtar MS,Akhtar M,et al. Edaravone protects rats against oxidative stress and apoptosis in experimentally induced myocardial infarction:Biochemical and ultrastructural evidence[J]. Redox Rep,2015,20(6):275-281.
[11] Janku F,Wheler JJ,Westin SN,et al. PI3K/AKT/m TOR inhibitors in patients with breast and gynecologic malignancies harboring PIK3CA mutations[J]. J Clin Oncol,2015,30(8):777-782.
[12] Chen K,Li G,Geng F,et al. Berberine reduces ischemia/reperfusion-induced myocardial apoptosis via activating AMPK and PI3K-Akt signaling in diabetic rats[J]. Apoptosis,2014,19(6):946-957.
[13]曹慧敏,宋囡,张妮,等.丹参酮Ⅱ_A通过PI3K/Akt/m TOR信号通路调控自噬抗内皮细胞氧化应激损伤研究[J].北京中医药大学学报,2017,40(11):933-939.
[14]金云晔,翟昌林,沈震,等.丹参酮ⅡA磺酸钠对大鼠缺血再灌注心肌细胞凋亡的影响[J].中华中医药学刊,2014,32(6):1414-1416.
[15]李佳. PI3K/AKT/GSK3-b信号转导通路在红景天苷保护大鼠心肌缺血再灌注损伤中的作用机制[J].中西医结合心脑血管病杂志,2016,14(22):2621-2624.
[16]韩军,宣佳利,胡浩然,等.金丝桃苷预处理减轻大鼠心肌缺血再灌注损伤作用与PI3K/Akt信号通路的关系[J].中国中药杂志,2015,40(1):118-123.
[17] Opferman JT. Attacking cancer’s Achilles heel:Antagonism of anti-apoptotic BCL-2 family members[J]. FEBS J,2016,283(14):2661-2675.
[18] Vasilikos L,Spilgies LM,Knop J,et al. Regulating the balance between necroptosis,apoptosis and inflammation by inhibitors of apoptosis proteins[J]. Immunol Cell Biol,2017,95(2):160-165.
[19]王厚磊,卢伟,李德芳,等.大鼠椎间盘髓核细胞体外凋亡模型的建立[J].中国实验动物学报,2015,23(6):607-611.