PPARγ/NF-κB信号通路参与替米沙坦抑制血管紧张素Ⅱ诱导的大鼠心肌纤维化及改善心功能作用
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摘要
目的观察替米沙坦对血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌纤维化及心脏功能的作用,并探讨其可能分子机制。方法 40只4周龄雄性SD大鼠随机分为4组,每组10只,分别为对照组(Vehicle/Veh)、血管紧张素Ⅱ组(AngⅡ)、血管紧张素Ⅱ+替米沙坦组(AngⅡ+Telm)和替米沙坦组(Telm)。采用皮下渗透压缓释泵持续泵入生理盐水(对照组)或AngⅡ(AngⅡ组)制作心肌纤维化大鼠模型,随后应用替米沙坦灌胃法对其进行干预,分别建立替米沙坦组和AngⅡ+替米沙坦组。分别应用M型超声心动图评价每组大鼠短轴缩短率(fraction shortening,FS),心脏血流动力学检查每组大鼠左室收缩压(left ventricular systolic pressure,LVSP)、左室舒张末压力(left ventricular end-diastolic pressure,LVEDP)以评估心脏功能;采用天狼猩红染色法评估心肌纤维化程度;为评估PPARγ/NF-κB信号通路的活化,采用蛋白免疫印迹法检测过氧化物增殖物激活受体γ(peroxisome proliferator activated receptor,PPARγ)、核因子-κB(nuclear factor-κB,NF-κB)蛋白水平,应用免疫荧光法检测细胞核NF-κB活化入核情况。结果与对照组大鼠心脏相比,AngⅡ组大鼠心脏FS显著下降(P <0. 05),LVSP显著下降(P <0. 05),LVEDP显著升高(P <0. 05);胶原表达显著增加(P <0. 05);组织PPARγ蛋白水平下调(P <0. 05),细胞核NF-κB蛋白水平上调(P <0. 05)且活化入核。与AngⅡ组相比,AngⅡ+替米沙坦组大鼠FS显著升高(P <0. 05),LVSP显著升高(P <0. 05),LVEDP显著下降(P <0. 05);胶原表达显著减少(P <0. 05);组织PPARγ蛋白水平上调(P <0. 05),细胞核NF-κB蛋白水平下调(P <0. 05)且活化入核受抑。与对照组相比,替米沙坦组大鼠心脏FS,LVSP,LVEDP,胶原表达量,组织PPARγ蛋白水平,细胞核NF-κB蛋白水平均无显著变化,NF-κB入核不明显。结论 PPARγ/NF-κB信号通路参与替米沙坦抑制血管紧张素Ⅱ诱导的大鼠心肌纤维化以改善心功能作用。
Objective To explore the effects of telmisartan on cardiac fibrosis and cardiac function in Ang Ⅱ-treated rat hearts and its underlying mechanisms. Methods Forty SD rats were randomly divided into four groups( n = 10 in each group) : vehicle group,AngⅡ group,Telm group and Ang Ⅱ + Telm group. Rats were given saline solution( vehicle group) or AngⅡ( Ang Ⅱ group) for 4 weeks via alzet osmotic mini-pumps,and then given telmisartan by gavage for 2 weeks in Telm group and Ang Ⅱ + Telm group. M-mode echocardiography was used for capturing fraction shortening( FS),and haemodynamic record was used for measuring the left ventricular systolic pressure( LVSP) and the left ventricular end-diastolic pressure( LVEDP). The cardiac fibrosis was analyzed by staining with sirius red. Western blot was used to detect the expression of peroxisome proliferator activated receptor( PPARγ) and nuclear factor( NF-κB) proteins,and immunofluorescence was performed to observe the activated NF-κB in nucleis. Results Compared with vehicle group,FS and LVSP significantly reduced in Ang Ⅱ group( P < 0. 05),LVEDP and collagen expression increased significantly( P< 0. 05),PPARγ protein expression was down-regulated and nuclear NF-κB protein expression was up-regulated significantly( P <0. 05),and the activated nuclear NF-κB displayed increasing. Compared with Ang Ⅱ group,FS and LVSP increased significantly in Ang Ⅱ + Telm group( P < 0. 05),LVEDP and collagen expression reduced significantly( P < 0. 05),PPARγ protein expression was up-regulated and nuclear NF-κB protein expression was down-regulated significantly( P < 0. 05),and the activated nuclear NF-κB displayed decreasing. All of FS,LVSP,LVEDP,collagen expression,PPARγ protein expression,NF-κB protein expression and NF-κB in nucleis showed no significant difference between Telm group and vehicle group. Conclusion PPARγ/NF-κB signal pathway may be involved in the anti-fibrotic and cardioprotective effects of telmisartan in Ang Ⅱ-treated rat hearts.
Objective To explore the effects of telmisartan on cardiac fibrosis and cardiac function in Ang Ⅱ-treated rat hearts and its underlying mechanisms. Methods Forty SD rats were randomly divided into four groups( n = 10 in each group) : vehicle group,AngⅡ group,Telm group and Ang Ⅱ + Telm group. Rats were given saline solution( vehicle group) or AngⅡ( Ang Ⅱ group) for 4 weeks via alzet osmotic mini-pumps,and then given telmisartan by gavage for 2 weeks in Telm group and Ang Ⅱ + Telm group. M-mode echocardiography was used for capturing fraction shortening( FS),and haemodynamic record was used for measuring the left ventricular systolic pressure( LVSP) and the left ventricular end-diastolic pressure( LVEDP). The cardiac fibrosis was analyzed by staining with sirius red. Western blot was used to detect the expression of peroxisome proliferator activated receptor( PPARγ) and nuclear factor( NF-κB) proteins,and immunofluorescence was performed to observe the activated NF-κB in nucleis. Results Compared with vehicle group,FS and LVSP significantly reduced in Ang Ⅱ group( P < 0. 05),LVEDP and collagen expression increased significantly( P< 0. 05),PPARγ protein expression was down-regulated and nuclear NF-κB protein expression was up-regulated significantly( P <0. 05),and the activated nuclear NF-κB displayed increasing. Compared with Ang Ⅱ group,FS and LVSP increased significantly in Ang Ⅱ + Telm group( P < 0. 05),LVEDP and collagen expression reduced significantly( P < 0. 05),PPARγ protein expression was up-regulated and nuclear NF-κB protein expression was down-regulated significantly( P < 0. 05),and the activated nuclear NF-κB displayed decreasing. All of FS,LVSP,LVEDP,collagen expression,PPARγ protein expression,NF-κB protein expression and NF-κB in nucleis showed no significant difference between Telm group and vehicle group. Conclusion PPARγ/NF-κB signal pathway may be involved in the anti-fibrotic and cardioprotective effects of telmisartan in Ang Ⅱ-treated rat hearts.
引文
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[19]Naveed M,Han L,Hasnat M,et al.Suppression of TGP on myocardial remodeling by regulating the NF-kappaB pathway[J].Biomed Pharmacother,2018,108(12):1460-1468.
[20]Suzuki S,Mori Y,Nagano A,et al.Pioglitazone,a peroxisome proliferator-activated receptor gamma agonist,suppresses rat prostate carcinogenesis[J].Int J Mol Sci,2016,17(12):2071-2081.
[21]Wan F,Lenardo MJ.The nuclear signaling of NF-kappaB:current knowledge,new insights,and future perspectives[J].Cell Res,2010,20(1):24-33.
[22]Merk DR,Chin JT,Dake BA,et al.mi R-29b participates in early aneurysm development in Marfan syndrome[J].Circ Res,2012,110(2):312-324.
[23]Penas FN,Cevey AC,Siffo S,et al.Hepatic injury associated with trypanosoma cruzi infection is attenuated by treatment with15-deoxy-Delta12,14prostaglandin J2[J].Exp Parasitol,2016,170(11):100-108.
[24]Zulueta A,Colombo M,Peli V,et al.Lung mesenchymal stem cells-derived extracellular vesicles attenuate the inflammatory profile of cystic fibrosis epithelial cells[J].Cell Signal,2018,51(11):110-118.
[25]Torres VE,Abebe KZ,Chapman AB,et al.Angiotensin blockade in late autosomal dominant polycystic kidney disease[J].NEngl J Med,2014,371(24):2267-2276.
[26]Whaley-Connell A,Habibi J,Panfili Z,et al.AngiotensinⅡactivation of m TOR results in tubulointerstitial fibrosis through loss of N-cadherin[J].Am J Nephrol,2011,34(2):115-125.
[27]Halici Z,Bilen H,Albayrak F,et al.Does telmisartan prevent hepatic fibrosis in rats with alloxan-induced diabetes?[J].Eur JPharmacol,2009,614(1-3):146-152.
[2]Brilla CG,Rupp H,Funck R,et al.The renin-angiotensin-aldosterone system and myocardial collagen matrix remodelling in congestive heart failure[J].Eur Heart J,1995,16(12):107-109.
[3]Ghazi L,Drawz P.Advances in understanding the renin-angiotensin-aldosterone system(RAAS)in blood pressure control and recent pivotal trials of RAAS blockade in heart failure and diabetic nephropathy[J].F1000res,2017,6(21):297-307.
[4]Akhrass PR,Mc Farlane SI.Telmisartan and cardioprotection[J].Vasc Health Risk Manag,2011,7(1):677-683.
[5]Kobayashi N,Ohno T,Yoshida K,et al.Cardioprotective mechanism of telmisartan via PPAR-gamma-eNOS pathway in dahl saltsensitive hypertensive rats[J].Am J Hypertens,2008,21(5):576-581.
[6]Sukumaran V,Watanabe K,Veeraveedu PT,et al.Telmisartan,an angiotensin-Ⅱreceptor blocker ameliorates cardiac remodeling in rats with dilated cardiomyopathy[J].Hypertens Res,2010,33(7):695-702.
[7]宋占春,白静慧,张丽莉,等.替米沙坦对大鼠急性心肌梗死后炎症反应及纤维化干预机制[J].中华医学杂志,2014,94(33):2628-2633.
[8]Chang WT,Cheng JT,Chen ZC.Telmisartan improves cardiac fibrosis in diabetes through peroxisome proliferator activated receptor delta(PPAR delta):from bedside to bench[J].Cardiovasc Diabetol,2016,15(1):113-121.
[9]Zhang Y,Zhao NA,Wang JK,et al.Telmisartan inhibited angiotensinⅡ-induced collagen metabolic imbalance without directly targeting TGF-beta 1/Smad signaling pathway in cardiac fibroblasts[J].Minerva Cardioangiol,2015,63(6):507-514.
[10]Samuel CS,Hewitson TD,Zhang Y,et al.Relaxin ameliorates fibrosis in experimental diabetic cardiomyopathy[J].Endocrinology,2008,149(7):3286-3293.
[11]Simpson P,Savion S.Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells.Cross-striations,ultrastructure,and chronotropic response to isoproterenol[J].Circ Res,1982,50(1):101-116.
[12]Tarbit E,Singh I,Peart JN,et al.Biomarkers for the identification of cardiac fibroblast and myofibroblast cells[J].Heart Fail Rev,2018,10(17):1-15.
[13]Leask A.Potential therapeutic targets for cardiac fibrosis:TGF-beta,angiotensin,endothelin,CCN2,and PDGF,partners in fibroblast activation[J].Circ Res,2010,106(11):1675-1680.
[14]Yu L,Yang G,Weng X,et al.Histone methyltransferase SET1mediates angiotensinⅡ-induced endothelin-1 transcription and cardiac hypertrophy in mice[J].Arterioscler Thromb Vasc.Biol,2015,35(5):1207-1217.
[15]Cui L,Wang Y,Yu R,et al.Jia-Shen decoction-medicated serum inhibits angiotensin-Ⅱinduced cardiac fibroblast proliferation via the TGF-beta1/Smad signaling pathway[J].Mol Med Rep,2016,14(2):1610-1616.
[16]Nair AR,Agbor LN,Mukohda M,et al.Interference with endothelial PPAR(peroxisome proliferator-activated receptor)-gamma causes accelerated cerebral vascular dysfunction in response to endogenous renin-angiotensin system activation[J].Hypertension,2018,72(5):1227-1235.
[17]Zhao N,Yu H,Yu H,et al.Mi RNA-711-SP1-collagen-Ⅰpathway is involved in the anti-fibrotic effect of pioglitazone in myocardial infarction[J].Sci China Life Sci,2013,56(5):431-439.
[18]Shen X,Li H,Li W,et al.Telmisartan ameliorates adipo R1 and adipoR2 expression via PPAR-gamma activation in the coronary artery and VSMCs[J].Biomed Pharmacother,2017,95(11):129-136.
[19]Naveed M,Han L,Hasnat M,et al.Suppression of TGP on myocardial remodeling by regulating the NF-kappaB pathway[J].Biomed Pharmacother,2018,108(12):1460-1468.
[20]Suzuki S,Mori Y,Nagano A,et al.Pioglitazone,a peroxisome proliferator-activated receptor gamma agonist,suppresses rat prostate carcinogenesis[J].Int J Mol Sci,2016,17(12):2071-2081.
[21]Wan F,Lenardo MJ.The nuclear signaling of NF-kappaB:current knowledge,new insights,and future perspectives[J].Cell Res,2010,20(1):24-33.
[22]Merk DR,Chin JT,Dake BA,et al.mi R-29b participates in early aneurysm development in Marfan syndrome[J].Circ Res,2012,110(2):312-324.
[23]Penas FN,Cevey AC,Siffo S,et al.Hepatic injury associated with trypanosoma cruzi infection is attenuated by treatment with15-deoxy-Delta12,14prostaglandin J2[J].Exp Parasitol,2016,170(11):100-108.
[24]Zulueta A,Colombo M,Peli V,et al.Lung mesenchymal stem cells-derived extracellular vesicles attenuate the inflammatory profile of cystic fibrosis epithelial cells[J].Cell Signal,2018,51(11):110-118.
[25]Torres VE,Abebe KZ,Chapman AB,et al.Angiotensin blockade in late autosomal dominant polycystic kidney disease[J].NEngl J Med,2014,371(24):2267-2276.
[26]Whaley-Connell A,Habibi J,Panfili Z,et al.AngiotensinⅡactivation of m TOR results in tubulointerstitial fibrosis through loss of N-cadherin[J].Am J Nephrol,2011,34(2):115-125.
[27]Halici Z,Bilen H,Albayrak F,et al.Does telmisartan prevent hepatic fibrosis in rats with alloxan-induced diabetes?[J].Eur JPharmacol,2009,614(1-3):146-152.