小凹蛋白-1介导阿托伐他汀对自发性高血压大鼠肾叶间动脉收缩反应的抑制作用
|
摘要
目的肾叶间动脉收缩反应增强是高血压肾脏损伤的早期表现,是重要的预防和治疗靶点。平滑肌细胞小凹蛋白-1(caveolin-1)上调是其发生机制之一,阿托伐他汀(atorvastatin,ATVS)可通过降低胆固醇(cholesterol,CHO)调控caveolin-1表达,本研究探讨ATVS是否通过caveolin-1抑制自发性高血压大鼠(spontaneously hypertensive rat,SHR)肾叶间动脉对血管紧张素II(angiotensin,Ang II)的收缩反应。方法 12只SHR大鼠随机分为2组:SHR对照组及ATVS(50 mg·kg~(-1)·d~(-1))处理组,每组6只,6只Wistar Kyoto(WKY)大鼠作为正常对照组。采用尾套法测量大鼠尾动脉收缩压(systolic blood pressure,SBP),血管环张力描记技术检测肾叶间动脉收缩反应,蛋白免疫印迹(Western blot)法检测caveolin-1和血管紧张素Ⅱ1型受体(angiotensin II type 1 receptor,AT1)蛋白表达,免疫沉淀和Western blot检测caveoin-1与AT1的结合。结果与WKY大鼠相比,SHR组SBP显著增高(P<0.05),肾叶间动脉对AngⅡ的收缩反应强度及敏感性均显著增强(P<0.05);4周ATVS处理显著降低SHR组SBP(P<0.05)及肾叶间动脉对AngⅡ的收缩反应强度及敏感性(P<0.05)。SHR组动脉AT1及caveolin-1的蛋白表达显著高于WKY大鼠(P<0.05),AngⅡ孵育所致caveolin-1与AT1的结合亦较WKY组显著增多(P<0.05)。ATVS处理可降低SHR组caveolin-1和AT1的蛋白表达(P<0.05),并抑制caveolin-1与AT1的结合(P<0.05),CHO孵育则可增高ATVS组caveolin-1蛋白含量并促进其与AT1的相互作用(P<0.05),同时可显著增加肾叶间动脉对AngⅡ收缩反应的强度及敏感性(P<0.05)。结论 ATVS可抑制SHR大鼠肾叶间动脉对AngⅡ的收缩反应,其机制与caveolin-1的蛋白表达降低及caveolin-1与AT1的结合减少有关。
Objective Excessive vasoconstriction of interlobar renal artery( IRA) is one of preliminary manifestations of renal damage in hypertension,during which caveolin-1 plays an important role. Atorvastatin( ATVS) represses cholesterol( CHO) synthesis to down-regulate the expression of caveolin-1,so we aimed to investigate whether the ATVS inhibited vasoconstriction of IRA to angiotensinⅡ( Ang Ⅱ) by regulating caveolin-1 in spontaneously hypertensive rats( SHR). Methods Twelve SHR were randomly divided into two groups: SHR control group,and SHR ATVS-treated group. Six Wistar Kyoto( WKY) rats weighing the same were employed as control group. Systolic blood pressure( SBP) was measured with the tail-cuff technique before and 2 or 4 weeks after ATVS treatment. Isometric force recording system was used to detect the vascular function. Western blotting was conducted toexamine the protein abundance of caveolin-1 or angiotensin Ⅱ type 1 receptor( AT_1). Immumoprecipitation was carried out to assess the binding of caveolin-1 and AT_1. Results SBP,and intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ were significantly increased in SHR as compared to WKY rats( P < 0. 05). At 4 th week,ATVS treatment could significantly reduce SPB( P < 0. 05),and the intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ( P < 0. 05) in SHR control group. Meanwhile,the protein expression of caveolin-1 or AT_1 as well as Ang Ⅱ elicited binding of caveolin-1 or AT_1 in IRA was higher in SHR than in WKY rats,which could also be reduced by ATVS( P < 0. 05).The ex vivo incubation of CHO raised the caveolin-1 protein content and promoted the binding of caveolin-1 and AT_1 in IRA of ATVS-treated SHR( P < 0. 05). At the same time,CHO enhanced the intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ( P < 0. 05). Conclusions ATVS mitigated the vasoconstriction of IRA to Ang Ⅱ in SHR,which was mediated by reducing caveolin-1 expression and inhibiting the binding of caveolin-1 and AT_1.
Objective Excessive vasoconstriction of interlobar renal artery( IRA) is one of preliminary manifestations of renal damage in hypertension,during which caveolin-1 plays an important role. Atorvastatin( ATVS) represses cholesterol( CHO) synthesis to down-regulate the expression of caveolin-1,so we aimed to investigate whether the ATVS inhibited vasoconstriction of IRA to angiotensinⅡ( Ang Ⅱ) by regulating caveolin-1 in spontaneously hypertensive rats( SHR). Methods Twelve SHR were randomly divided into two groups: SHR control group,and SHR ATVS-treated group. Six Wistar Kyoto( WKY) rats weighing the same were employed as control group. Systolic blood pressure( SBP) was measured with the tail-cuff technique before and 2 or 4 weeks after ATVS treatment. Isometric force recording system was used to detect the vascular function. Western blotting was conducted toexamine the protein abundance of caveolin-1 or angiotensin Ⅱ type 1 receptor( AT_1). Immumoprecipitation was carried out to assess the binding of caveolin-1 and AT_1. Results SBP,and intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ were significantly increased in SHR as compared to WKY rats( P < 0. 05). At 4 th week,ATVS treatment could significantly reduce SPB( P < 0. 05),and the intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ( P < 0. 05) in SHR control group. Meanwhile,the protein expression of caveolin-1 or AT_1 as well as Ang Ⅱ elicited binding of caveolin-1 or AT_1 in IRA was higher in SHR than in WKY rats,which could also be reduced by ATVS( P < 0. 05).The ex vivo incubation of CHO raised the caveolin-1 protein content and promoted the binding of caveolin-1 and AT_1 in IRA of ATVS-treated SHR( P < 0. 05). At the same time,CHO enhanced the intensity and sensitivity of vasoconstriction of IRA to Ang Ⅱ( P < 0. 05). Conclusions ATVS mitigated the vasoconstriction of IRA to Ang Ⅱ in SHR,which was mediated by reducing caveolin-1 expression and inhibiting the binding of caveolin-1 and AT_1.
引文
[1]Tozawa M,lseki K,Iseki C,et al.Blood pressure predicts risk of developing en-stage renal disease in men and women[J].Hypertension,2003,41(6):1341-1345.
[2]Protasiewicz M,Kadziela J,Poczatek K,et al.Renal artery stenosis in patients with resistant hypertension[J].Am J Cardiol,2013,112(9):1417-1420.
[3]Te RL,van Esch JH,Roks AJ,et al.Hypertension:renin-angiotensin-aldosterone system alterations[J].Circ Res,2015,116(6):960-975.
[4]王双,万载阳,唐朝克,等.两肾一夹型高血压大鼠重塑血管对血管紧张素Ⅱ反应变化及其机制[J].中国动脉硬化杂志,2003,11(6):501-504.
[5]张海军,张茜,谢小萍,等.平滑肌细胞caveolae在自发性高血压大鼠肾叶间动脉收缩功能改变中的作用研究[J].临床肾脏病杂志,2016,16(10):625-631.
[6]张海军,苏玉婷,孟星星,等.平滑肌细胞caveolae在大鼠肾动脉收缩反应调控中的作用研究[J].临床肾脏病杂志,2016,16(7):432-438.
[7]Callera GE,Montezano AC,Yogi A,et al.Vascular signaling through cholesterol-rich domains:implications in hypertension[J].Curr Opin Nephrol Hypertens,2007,16(2):90-104.
[8]Wang Z,Bai Y,Yu J,et al.Caveolae regulate vasoconstriction of conduit arteries to angiotensinⅡin hindlimb unweighted rats[J].J Physiol,2015,593(20):4561-4574.
[9]Razani B,Lisanti MP.Caveolins and caveolae:molecular and functional relationships[J].Exp Cell Res,2001,271(1):36-44.
[10]Krane V,Wanner C.Statins,inflammation and kidney disease[J].Nat Rev Nephrol,2011,7(7):385-397.
[11]谢晓竞,杨解人,王安才.阿托伐他汀的降血压作用及其机制实验研究[J].中华全科医学,2008,6(9):882-884,封3.
[12]Taylor FC,Huffman M,Ebrahim S.Statin therapy for primary prevention of cardiovascular disease[J].JAMA,2013,310(22):2451-2452.
[13]郑颖,吕明睿.阿托伐他汀改善高血压患者早期肾损害的研究[J].中外健康文摘,2012,40:132-133.
[14]Sinatra ST,Teter BB,Bowden J,et al.The cholesterol and statin controversy:the new 2013 statin-cholesterol guidelines[J].Altern Ther Health Med,2014,20(5):14-17.
[15]Bidani AK,Griffin KA.Pathophysiology of hypertensive renal damage:implications for therapy[J].Hypertension,2004,44(5):595-601.
[16]许红强,王金萍,常丽娜.肾叶间动脉阻力指数与原发性高血压早期肾损害的相关性[J].安徽中医学院学报,2006,25(4):51-52.
[17]张雯,马克涛,刘卫东,等.缝隙连接在自发性高血压大鼠肾叶间动脉收缩增强中的作用[J].安徽医科大学学报,2014,49(11):1570-1573.
[18]Burnier M,Brunner HR.AngiotensinⅡreceptor antagonists[J].Lancet,2000,355(9204):637-645.
[19]阮志芹,李华.原发性高血压和肾功能减退的相关性[J].中华高血压杂志,2017,1:22-26.
[20]张志杰,胡申江,孙坚,等.阿托伐他汀对自发性高血压大鼠血管内皮功能的保护作用[J].浙江大学学报(医学版),2007,36(4):355-359.
[21]Li CL,Chou HW,Chan KA,et al.Effects of atorvastatin and rosuvastatin on renal function in patients with type 2 diabetes mellitus[J].Am J Cardiol,2015,115(5):619-624.
[22]Chang CH,Lee YC,Tsai CT,et al.Continuation of statin therapy and a decreased risk of atrial fibrillation/flutter in patients with and without chronic kidney disease[J].Atherosclerosis,2014,232(1):224-230.
[23]焦海霞,李凌,余涓,等.尾套法和颈动脉法测定急性应激大鼠血压变化值的比较[J].福建医科大学学报,2006,40(4):404-405.
[24]张勤勤,孙润广,李连启.Caveolae及其蛋白家族的生物学结构和功能的研究现状[J].北京生物医学工程,2011,30(2):215-220.
[25]Yin X,Li B,Chen H,et al.Differential signaling pathways in angiotensinⅡ-and epidermal growth factor-stimulated hepatic C9cells[J].Mol Pharmacol,2008,74(5):1223-1233.
[26]Linder AE,Thakali KM,Thompson JM,et al.Methyl-beta-cyclodextrin prevents angiotensinⅡ-induced tachyphylactic contractile responses in rat aorta[J].J Pharmacol Exp Ther,2007,323(1):78-84.
[27]Busija AR,Patel HH,Insel PA.Caveolins and cavins in the trafficking,maturation,and degradation of caveolae:implications for cell physiology[J].Am J Physiol Cell Physiol,2017,312(4):C459-C477.
[28]Raghow R.Statins redux:a re-assessment of how statins lower plasma cholesterol[J].World J Diabetes,2017,8(6):230-234.
[29]Guasti L,Marino F,Cosentino M,et al.Prolonged statin-associated reduction in neutrophil reactive oxygen species and angiotensinⅡtype 1 receptor expression:1-year follow-up[J].Eur Heart J,2008,29(9):1118-1126.
[30]Nickenig G,Baumer AT,Temur Y,et al.Statin-sensitive dysregulated AT1 receptor function and density in hypercholesterolemic men[J].Circulation,1999,100(21):2131-2134.
[2]Protasiewicz M,Kadziela J,Poczatek K,et al.Renal artery stenosis in patients with resistant hypertension[J].Am J Cardiol,2013,112(9):1417-1420.
[3]Te RL,van Esch JH,Roks AJ,et al.Hypertension:renin-angiotensin-aldosterone system alterations[J].Circ Res,2015,116(6):960-975.
[4]王双,万载阳,唐朝克,等.两肾一夹型高血压大鼠重塑血管对血管紧张素Ⅱ反应变化及其机制[J].中国动脉硬化杂志,2003,11(6):501-504.
[5]张海军,张茜,谢小萍,等.平滑肌细胞caveolae在自发性高血压大鼠肾叶间动脉收缩功能改变中的作用研究[J].临床肾脏病杂志,2016,16(10):625-631.
[6]张海军,苏玉婷,孟星星,等.平滑肌细胞caveolae在大鼠肾动脉收缩反应调控中的作用研究[J].临床肾脏病杂志,2016,16(7):432-438.
[7]Callera GE,Montezano AC,Yogi A,et al.Vascular signaling through cholesterol-rich domains:implications in hypertension[J].Curr Opin Nephrol Hypertens,2007,16(2):90-104.
[8]Wang Z,Bai Y,Yu J,et al.Caveolae regulate vasoconstriction of conduit arteries to angiotensinⅡin hindlimb unweighted rats[J].J Physiol,2015,593(20):4561-4574.
[9]Razani B,Lisanti MP.Caveolins and caveolae:molecular and functional relationships[J].Exp Cell Res,2001,271(1):36-44.
[10]Krane V,Wanner C.Statins,inflammation and kidney disease[J].Nat Rev Nephrol,2011,7(7):385-397.
[11]谢晓竞,杨解人,王安才.阿托伐他汀的降血压作用及其机制实验研究[J].中华全科医学,2008,6(9):882-884,封3.
[12]Taylor FC,Huffman M,Ebrahim S.Statin therapy for primary prevention of cardiovascular disease[J].JAMA,2013,310(22):2451-2452.
[13]郑颖,吕明睿.阿托伐他汀改善高血压患者早期肾损害的研究[J].中外健康文摘,2012,40:132-133.
[14]Sinatra ST,Teter BB,Bowden J,et al.The cholesterol and statin controversy:the new 2013 statin-cholesterol guidelines[J].Altern Ther Health Med,2014,20(5):14-17.
[15]Bidani AK,Griffin KA.Pathophysiology of hypertensive renal damage:implications for therapy[J].Hypertension,2004,44(5):595-601.
[16]许红强,王金萍,常丽娜.肾叶间动脉阻力指数与原发性高血压早期肾损害的相关性[J].安徽中医学院学报,2006,25(4):51-52.
[17]张雯,马克涛,刘卫东,等.缝隙连接在自发性高血压大鼠肾叶间动脉收缩增强中的作用[J].安徽医科大学学报,2014,49(11):1570-1573.
[18]Burnier M,Brunner HR.AngiotensinⅡreceptor antagonists[J].Lancet,2000,355(9204):637-645.
[19]阮志芹,李华.原发性高血压和肾功能减退的相关性[J].中华高血压杂志,2017,1:22-26.
[20]张志杰,胡申江,孙坚,等.阿托伐他汀对自发性高血压大鼠血管内皮功能的保护作用[J].浙江大学学报(医学版),2007,36(4):355-359.
[21]Li CL,Chou HW,Chan KA,et al.Effects of atorvastatin and rosuvastatin on renal function in patients with type 2 diabetes mellitus[J].Am J Cardiol,2015,115(5):619-624.
[22]Chang CH,Lee YC,Tsai CT,et al.Continuation of statin therapy and a decreased risk of atrial fibrillation/flutter in patients with and without chronic kidney disease[J].Atherosclerosis,2014,232(1):224-230.
[23]焦海霞,李凌,余涓,等.尾套法和颈动脉法测定急性应激大鼠血压变化值的比较[J].福建医科大学学报,2006,40(4):404-405.
[24]张勤勤,孙润广,李连启.Caveolae及其蛋白家族的生物学结构和功能的研究现状[J].北京生物医学工程,2011,30(2):215-220.
[25]Yin X,Li B,Chen H,et al.Differential signaling pathways in angiotensinⅡ-and epidermal growth factor-stimulated hepatic C9cells[J].Mol Pharmacol,2008,74(5):1223-1233.
[26]Linder AE,Thakali KM,Thompson JM,et al.Methyl-beta-cyclodextrin prevents angiotensinⅡ-induced tachyphylactic contractile responses in rat aorta[J].J Pharmacol Exp Ther,2007,323(1):78-84.
[27]Busija AR,Patel HH,Insel PA.Caveolins and cavins in the trafficking,maturation,and degradation of caveolae:implications for cell physiology[J].Am J Physiol Cell Physiol,2017,312(4):C459-C477.
[28]Raghow R.Statins redux:a re-assessment of how statins lower plasma cholesterol[J].World J Diabetes,2017,8(6):230-234.
[29]Guasti L,Marino F,Cosentino M,et al.Prolonged statin-associated reduction in neutrophil reactive oxygen species and angiotensinⅡtype 1 receptor expression:1-year follow-up[J].Eur Heart J,2008,29(9):1118-1126.
[30]Nickenig G,Baumer AT,Temur Y,et al.Statin-sensitive dysregulated AT1 receptor function and density in hypercholesterolemic men[J].Circulation,1999,100(21):2131-2134.