泽泻汤加味方对盐敏感性HBZY21细胞中AngⅡ-NADPH-ROS信号通路的影响
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摘要
目的探讨泽泻汤加味方对盐敏感性肾小球系膜细胞中AngⅡ-NADPH-ROS信号通路的作用机制。方法采用高盐和AngⅡ诱导大鼠肾小球系膜细胞的方法建立盐敏感性高血压体外细胞模型,设正常组、模型组、阳性药缬沙坦组、泽泻汤加味方(高、中、低剂量)组。CKK-8法测定细胞活性;RT-qPCR方法检测细胞中Agtr1a、Cyba、NOX4的m RNA表达水平;Western blot方法检测细胞中AT1R、P22、P47、NOX4的蛋白质表达水平;活性氧检测试剂盒检测细胞中ROS水平。结果与正常组比较,模型组细胞中Agtr1a、Cyba、NOX4的mRNA表达水平、AT1R、P22、P47、NOX4蛋白表达水平以及ROS表达水平显著升高(P<0.05)。与模型组比较,缬沙坦组及泽泻汤加味方中剂量组Agtr1a、Cyba、NOX4的mRNA表达水平、AT1R、P22、P47、NOX4蛋白表达水平以及ROS表达水平显著降低(P<0.05)。不同浓度泽泻汤加味方组间,中剂量组对AngⅡ-NADPH-ROS信号通路的下调作用最为明显。结论泽泻汤加味方对盐敏感性高血压肾损害的保护作用可能与抑制AngⅡ-NADPH-ROS信号通路有关。
Objective To investigate the effect of modified Zexie Decoction on the angiotensin Ⅱ(AngⅡ)-NADPH-reactive oxygen species(ROS) signaling pathway in salt-sensitive glomerular mesangial cells. Methods An in vitro cell model of salt-sensitive hypertension was established by inducing rat glomerular mesangial cells with high salt and AngⅡ. Cells were divided into normal group, model group, positive valsartan group, and high-, medium-, and low-dose modified Zexie Decoction groups.CKK-8 assay was used to determine the cell viability. RT-PCR was used to determine the mRNA expression levels of Agtr1 a,Cyba, and NOX4. Western blot was used to determine the protein expression levels of AT1 R, P22, P47, and NOX4 in cells. The ROS levels in cells were measured by a ROS assay kit. Results Compared with the normal group, the model group had significantly higher mRNA levels of Agtr1 a, Cyba, NOX4, protein levels of AT1 R, P22, P47, and NOX4, and ROS levels(P<0.05).Compared with the model group, the valsartan group and the medium-dose modified Zexie Decoction group had significantly lower mRNA levels of Agtr1 a, Cyba, and NOX4, protein levels of AT1 R, P22, P47, and NOX4, and ROS levels(P<0.05). Among the three modified Zexie Decoction groups, the medium-dose group had the greatest downregulation of the AngⅡ-NADPH-ROS signaling pathway. Conclusion The protective effect of modified Zexie Decoction against renal injury induced by salt-sensitive hypertension might relate to the inhibition of the AngⅡ-NADPH-ROS signaling pathway.
Objective To investigate the effect of modified Zexie Decoction on the angiotensin Ⅱ(AngⅡ)-NADPH-reactive oxygen species(ROS) signaling pathway in salt-sensitive glomerular mesangial cells. Methods An in vitro cell model of salt-sensitive hypertension was established by inducing rat glomerular mesangial cells with high salt and AngⅡ. Cells were divided into normal group, model group, positive valsartan group, and high-, medium-, and low-dose modified Zexie Decoction groups.CKK-8 assay was used to determine the cell viability. RT-PCR was used to determine the mRNA expression levels of Agtr1 a,Cyba, and NOX4. Western blot was used to determine the protein expression levels of AT1 R, P22, P47, and NOX4 in cells. The ROS levels in cells were measured by a ROS assay kit. Results Compared with the normal group, the model group had significantly higher mRNA levels of Agtr1 a, Cyba, NOX4, protein levels of AT1 R, P22, P47, and NOX4, and ROS levels(P<0.05).Compared with the model group, the valsartan group and the medium-dose modified Zexie Decoction group had significantly lower mRNA levels of Agtr1 a, Cyba, and NOX4, protein levels of AT1 R, P22, P47, and NOX4, and ROS levels(P<0.05). Among the three modified Zexie Decoction groups, the medium-dose group had the greatest downregulation of the AngⅡ-NADPH-ROS signaling pathway. Conclusion The protective effect of modified Zexie Decoction against renal injury induced by salt-sensitive hypertension might relate to the inhibition of the AngⅡ-NADPH-ROS signaling pathway.
引文
[1]中国高血压防治指南修订委员会.中国高血压防治指南2010[J].中华高血压杂志,2011,19(8):707-743.
[2]王丽,卢成志,张欣,等.经皮肾脏交感神经射频消融术对顽固性高血压患者肾素血管紧张系统的影响[J].中华心血管病杂志,2013,41(1):3-7.
[3] VON LUEDER T G, KRUM H. RAAS inhibitors and cardiovascular protection in large scale trials[J]. Cardiovasc Drugs Ther, 2013,27(2):171-179.
[4] PRAJAPATI H, MCCALLUM A, FINLAY E. Hypertension, secondary to a renal artery aneurysm, treated by ex vivo aneurysm repair and autotrans plantation[J]. BMJ Case Reports, 2012,19(6):1725-1726.
[5]曾勇,张稳,叶舒婷,等.中西药物对高血压病炎症反应干预作用的研究进展[J].中西医结合心脑血管病杂志,2014;12(1):85-87.
[6]王超,孙磊,王振滔,等.联合中药对高血压治疗的研究进展[J].吉林医药学院学报,2018,39(4):290-292.
[7]范洪亮,张树峰,张连和,等.泽泻汤加味方治疗高血压80例降压效果临床观察[J].中国心脏与心律电子杂志,2016,4(1):3-6.
[8]张婷婷,蒋希成,吴鑫宇,等.泽泻汤加味方对高盐条件下Dahl大鼠高血压调节及肾功能的保护作用[J].中医药信息,2017,34(3):44-46.
[9]何红梅,梅爱敏,田河林,等.泽泻汤加味方对高盐诱导大鼠高血压肾功能损害的保护作用[J].山东医药,2017,57(17):31-33.
[10]韩运峰,苏诚坚,区碧如.非洛地平缓释片及缬沙坦对感觉神经损伤性盐敏感性高血压大鼠的降压作用及其机制探讨[J].中华心血管病杂志,2005,33(3):255-259.
[11] WOLAK T, ALIEV E, ROGACHEV B, et al. Renal safety and angiogenesis II blockade medications in patients undergoing nonemergent coronary angiography:a randomized controlled study[J].Israel Medical Association Journal, 2013,5(11):682-687.
[12] TUCKER S, CHEN Y, ABELL R. In patients with chronic diabetic nephropathy, do angiotensin converting enzyme inhibitors(ACEI)have greater renal protective effect as compared to angiotensin receptor blockers(ARB)[J]. J Okla State Med Assoc,2013,106(7):294-295.
[13] KUSHIRO T, FUJITA H, HISAKI R, et al. Oxidative stress in the Dahl salt-sensitive hypertensive rat[J]. Clinical and Experimental Hypertension, 2005,27(1):9-15.
[14] LIU L, WU X, XU H, et al. Myocardin-related transcription factor A(MRTF-A)contributes to acute kidney injury by regulating macrophage ROS production[J]. Biochim et Biophysica Acta(BBA)-Molecular Basis of Disease, 2018, 1864(10):3109-3121.
[15]简怡娟,程锦国,董飞侠.NOX通过ROS介导促肾纤维化作用[J].中国医师杂志,2015,17(7):1108-1110.
[16] ARDIANA F, SUCIATI, INDRAYANTO G. Valsartan[J]. Profiles of Drug Substances, Excipients and Related Methodology, 2015,40:431-493.
[17]冯柏,王屏,宁彰.缬沙坦致不良反应74例文献分析[J].中国新药杂志,2010,19(12):1094-1096.
[18]陈景彦,陈炳宏,范洪亮,等.泽泻汤加味方对高盐高血压肾损害大鼠AT1R、AT2R m RNA表达的影响[J].中医杂志,2016,57(5):428-430.
[2]王丽,卢成志,张欣,等.经皮肾脏交感神经射频消融术对顽固性高血压患者肾素血管紧张系统的影响[J].中华心血管病杂志,2013,41(1):3-7.
[3] VON LUEDER T G, KRUM H. RAAS inhibitors and cardiovascular protection in large scale trials[J]. Cardiovasc Drugs Ther, 2013,27(2):171-179.
[4] PRAJAPATI H, MCCALLUM A, FINLAY E. Hypertension, secondary to a renal artery aneurysm, treated by ex vivo aneurysm repair and autotrans plantation[J]. BMJ Case Reports, 2012,19(6):1725-1726.
[5]曾勇,张稳,叶舒婷,等.中西药物对高血压病炎症反应干预作用的研究进展[J].中西医结合心脑血管病杂志,2014;12(1):85-87.
[6]王超,孙磊,王振滔,等.联合中药对高血压治疗的研究进展[J].吉林医药学院学报,2018,39(4):290-292.
[7]范洪亮,张树峰,张连和,等.泽泻汤加味方治疗高血压80例降压效果临床观察[J].中国心脏与心律电子杂志,2016,4(1):3-6.
[8]张婷婷,蒋希成,吴鑫宇,等.泽泻汤加味方对高盐条件下Dahl大鼠高血压调节及肾功能的保护作用[J].中医药信息,2017,34(3):44-46.
[9]何红梅,梅爱敏,田河林,等.泽泻汤加味方对高盐诱导大鼠高血压肾功能损害的保护作用[J].山东医药,2017,57(17):31-33.
[10]韩运峰,苏诚坚,区碧如.非洛地平缓释片及缬沙坦对感觉神经损伤性盐敏感性高血压大鼠的降压作用及其机制探讨[J].中华心血管病杂志,2005,33(3):255-259.
[11] WOLAK T, ALIEV E, ROGACHEV B, et al. Renal safety and angiogenesis II blockade medications in patients undergoing nonemergent coronary angiography:a randomized controlled study[J].Israel Medical Association Journal, 2013,5(11):682-687.
[12] TUCKER S, CHEN Y, ABELL R. In patients with chronic diabetic nephropathy, do angiotensin converting enzyme inhibitors(ACEI)have greater renal protective effect as compared to angiotensin receptor blockers(ARB)[J]. J Okla State Med Assoc,2013,106(7):294-295.
[13] KUSHIRO T, FUJITA H, HISAKI R, et al. Oxidative stress in the Dahl salt-sensitive hypertensive rat[J]. Clinical and Experimental Hypertension, 2005,27(1):9-15.
[14] LIU L, WU X, XU H, et al. Myocardin-related transcription factor A(MRTF-A)contributes to acute kidney injury by regulating macrophage ROS production[J]. Biochim et Biophysica Acta(BBA)-Molecular Basis of Disease, 2018, 1864(10):3109-3121.
[15]简怡娟,程锦国,董飞侠.NOX通过ROS介导促肾纤维化作用[J].中国医师杂志,2015,17(7):1108-1110.
[16] ARDIANA F, SUCIATI, INDRAYANTO G. Valsartan[J]. Profiles of Drug Substances, Excipients and Related Methodology, 2015,40:431-493.
[17]冯柏,王屏,宁彰.缬沙坦致不良反应74例文献分析[J].中国新药杂志,2010,19(12):1094-1096.
[18]陈景彦,陈炳宏,范洪亮,等.泽泻汤加味方对高盐高血压肾损害大鼠AT1R、AT2R m RNA表达的影响[J].中医杂志,2016,57(5):428-430.
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