先兆子痫患者体内的抗体通过激活血管紧张素受体刺激细胞内钙离子动员增加

背景:先兆子痫是妊娠时发生的,以高血压、蛋白尿、水肿以及凝血和血管异常为特征的一种严重疾病。在细胞水平上,血小板、淋巴细胞及红细胞内钙离子浓度异常增加。新近的研究表明,针对血管紧张素Ⅱ1型受体(AT1)的抗体也与先兆子痫高度相关。方法和结果:验证血管紧张素Ⅱ1型受体激动样抗体(AT1-AAs)是否可激活AT1受体,从而导致细胞内游离钙浓度增加及下游Ca2+信号转导通路激活。采集30例妊娠者的血清,其中16例为重症先兆子痫,14例血压正常,检测能够激活细胞内钙离子动员的IgG。在所有先兆子痫患者中,IgG均可激活AT1受体,并增加细胞内游…     

碘化N-正丁基氟哌啶醇抑制血管紧张素Ⅱ刺激血管平滑肌细胞ERK1/2和cAMP反应元件结合蛋白的表达

目的:探讨碘化N-正丁基氟哌啶醇(F2)对血管紧张素Ⅱ(AngⅡ)刺激血管平滑肌细胞(VSMCs)ERK1/2和cAMP反应元件结合(CREB)蛋白表达的影响。方法:原代培养大鼠胸主动脉VSMCs;用Western blot法检测ERK1/2,p-ERK1/2,CREB和p-CREB蛋白表达。结果:AngⅡ可增加VSMCsp-ERK1/2和p-CREB蛋白表达,而不增加ERK1/2和CREB蛋白表达;F2(10-8,10-7和10-6mol/L)剂量-依赖地降低AngⅡ刺激的p-ERK1/2和p-CREB蛋白表达。结论:F2抑制AngⅡ刺激VSMCsERK1/2和CREB蛋白磷酸化,这可能是其抑制VSMCs增殖的机制。     

血管紧张素Ⅱ对肝癌细胞增殖的影响及其机制研究

目的:研究血管紧张素Ⅱ对肝癌HepG2细胞增殖的影响及其机制.方法:采用CCK-8法测定不同浓度血管紧张素Ⅱ作用24、48 h对HepG2细胞增殖的影响.通过蛋白质免疫印迹法测定血管紧张素Ⅱ处理后HepG2细胞内细胞外信号调节激酶(ERK)1/2和血管紧张素Ⅱ-1型受体(AT1R)蛋白表达水平.结果:作用24 h,10...     

ERK1/2信号通路在OX-LDL诱导的血管平滑肌细胞TLR4 mRNA表达中的作用

目的探讨ERK1/2信号通路在氧化性低密度脂蛋白（OX-LDL）诱导的血管平滑肌细胞（VSMCs）Toll样受体-4（TLR4）mRNA表达中的作用。方法采用贴块法培养大鼠VSMCs,在氧化低密度脂蛋白（OX-LDL）及PD98059（ERK1/2特异性抑制剂）作用下采用RT-PCR检测VSMCs TLR4 mRNA的表达,用Westernblotting检测ERK1/2磷酸化水平的变化。结果OX-LDL使VSMCs ERK1/2磷酸化水平升高;PD98059抑制ERK1/2的磷酸化;OX-LDL刺激VSMCs上调TLR4 mRNA的表达（P〈0.05）;PD98059预孵育后TLR4 mRNA的表达较单独OX-LDL刺激情况下降低（P〈0.05）。结论OX-LDL通过或部分通过ERK1/2信号通路介导VSMCs TLR4 mRNA的表达。     

AT1-AA和sFlt-1与子痫前期的关系

血管紧张素Ⅱ1型受体自身抗体(AT1-AA)通过类血管紧张素Ⅱ的激动效应,激活活性氧簇和炎性反应,使滋养细胞侵袭变浅,胎盘血流量减少和胎盘血管硬化。可溶性血管内皮生长因子受体1(sFlt-1)使机体处于抗血管发生状态,滋养细胞浅植入和不正常的子宫胎盘血管抵抗。AT1-AA可能促进sFlt-1的产生,共同参与子痫前期的发病。深入研究AT1-AA和sFlt-1的相关性,对了解子痫前期的发病机制及预测、预防具有十分重要的意义。     

Mfn2介导缬沙坦抑制血管平滑肌细胞增殖的研究

目的 研究缬沙坦对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的大鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的影响,并探讨其新的作用机制.方法 体外培养VSMCs,采用AngⅡ诱导其增殖,用不同浓度的缬沙坦(10-5、10-6、10-7mol/L)进行干预,细胞计数及四氮唑盐试验(MTT)检测VSMCs增殖情况,Western blot检测各组线粒体融合素基因-2(mitofusin-2,Mfn2)、Raf、细胞外信号调节激酶1/2(extracellular signalregulated kinase 1/2,ERK1/2)表达水平的变化.结果 AngⅡ能够明显促进VSMCs的增殖,下调Mfn2的表达、增强Raf和ERK1/2的表达;缬沙坦10-5、10-6mol/L可以拮抗AngⅡ的上述作用,而缬沙坦10-7mol/L无此作用;缬沙坦对无AngⅡ刺激的VSMCs增殖无明显影响.结论缬沙坦能有效抑制AngⅡ诱导的VSMCs增殖,其机制与调节Mfn2的表达、抑制Ras-Raf-ERK/MAPK信号通路有关.     

新型AT_1受体拮抗剂化合物Ib对血管紧张素Ⅱ诱导的血管平滑肌细胞增殖的抑制作用

目的:研究新型AT1受体拮抗剂化合物Ib对血管紧张素Ⅱ(AngⅡ)诱导的血管平滑肌细胞(VSMCs)增殖的抑制作用.方法:采用MTT法和3H-TdR掺入法测定AngⅡ促进VSMCs增殖作用;采用放射性受体配体分析法测定化合物Ib对VSMCs表面AT1受体的作用;用激光共聚焦显微镜观察化合物Ib对AngⅡ诱导的[ca2+];升高的拮抗作用.结果:化合物nb(0.1,0.5,2.5 μmol/L)显著抑制AngⅡ诱导的VSMCs增殖作用;化合物Ib 剂量相关性的抑制125I-Ang lI与VSMCs上AT1受体的结合,IC50为0.96 nmol/L;化合物lb(O.1,0.5,2.5 μmol/L)对AngⅡ诱导的[Ca22+];升高有显著的抑制作用.结论:化合物Ib可以抑制Angll诱导的VSMCs增殖作用,其可能的作用机制是通过拮抗ATl受体,进而抑制AngII诱导的[Ca2+];升高效应. 对AngⅡ诱导的[ca2+];升高的拮抗作用.结果:化合物nb(0.1,0.5,2.5 μmol/L)显著抑制AngⅡ诱导的VSMCs增殖作用;化合物Ib 剂量相关性的抑制125I-Ang lI与VSMCs上AT1受体 结合,IC50为0.96 nmol/L;化合物lb(O.1,0.5,2.5 μmol/L)对AngⅡ诱导的[Ca22+];升高有显著的抑制作用.结论:化合物Ib可以抑制Angll诱导的VSM     

血管紧张素Ⅱ2型受体对血管和肾脏功能的影响

肾素-血管紧张素系统(RAS)是机体调节血压和肾脏排钠的重要功能系统,其效应分子血管紧张素Ⅱ主要包括两种受体血管紧张素Ⅱ1型受体(AT1)和血管紧张素Ⅱ2型受体(AT2)。血管紧张素Ⅱ的主要功能是通过AT1受体介导的。现在越来越多的研究表明AT2受体有拮抗AT1受体的作用主要表现在血管舒张方面。近几年来研究表明AT2受体的微血管舒张作用是通过缓激肽相关或无关的方式产生NO来介导。虽然AT2受体在肾脏的近端小管、远端小管和血管系统都存在表达,AT2受体刺激诱导肾脏的缓激肽(bradykinin)/NO通道,但很少有关AT2受体调节肾功能和钠外排方面有价值的研究。现就近年来对AT2受体在血管舒张和肾功能的影响方面的最新研究进展简要综述。     

血管紧张素Ⅱ及其受体拮抗剂对肝星状细胞Ⅲ型胶原合成的影响

目的 观察血管紧张素Ⅱ及其受体拮抗剂对体外培养的肝星状细胞Ⅲ型胶原蛋白合成及其mRNA表达的影响.方法 采用HSC-T6肝星状细胞系作为活化的肝星状细胞的研究模型.将培养的肝星状细胞随机分为对照组、血管紧张素Ⅱ(AngⅡ)组、受体拮抗剂(AT1RA)组和血管紧张素Ⅱ 受体拮抗剂(AngⅡ AT1RA)组.采用ELISA法检测细胞培养上清液中Ⅲ型胶原蛋白的含量;RT-PCR法检测肝星状细胞中Ⅲ型胶原mRNA的表达.结果 细胞培养上清液中Ⅲ型胶原蛋白的含量对照组、AngⅡ组和AngⅡ AT1RA组分别为(6.301±0.432)ng/ml、(7.356±0.237)ng/ml和(6.263±0.236)ng/ml,AngⅡ组高于对照组(P＜0.05),AngⅡ AT1RA组显著低于AngⅡ组(P＜0.05).肝星状细胞Ⅲ型胶原mRNA的表达水平对照组、AngⅡ组和AngⅡ AT1RA组分别为2.317±0.015、2.643±0.057和2.330±0.036,AngⅡ组高于对照组(P＜0.05),AngⅡ AT1RA组显著低于AngⅡ组(P＜0.05).结论 血管紧张素Ⅱ能够促进肝星状细胞Ⅲ型胶原蛋白的合成及其mRNA的表达,而血管紧张素Ⅱ1型受体拮抗剂能够明显抑制这一作用.     

血管紧张素Ⅱ及其受体在疼痛中的研究进展

血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)是肾素-血管紧张素系统(the renin-angiotensin system,RAS)的主要活性肽,主要作用于AT1R(angiotensin Ⅱ receptor)和AT2R(angiotensin Ⅱ type 2 receptor)两种受体,在人类及大鼠组织中广泛表达。近年来发现AngⅡ及其受体还参与疼痛的调节,其受体拮抗剂可应用与临床疼痛的治疗。本文就血管紧张素Ⅱ及其受体在疼痛中的研究进展进行综述。     

Effects of Cyclosporine A on Angiotensin Ⅱ-induced Cardiomyocyte Hypertrophy in Neonatal Rats

The effects of cyclosporine A (CsA) on Angiontensin Ⅱ (Ang Ⅱ )-induced protein contents, c-fos protein levels and cytosolic Ca2+ level ([Ca2+ ]i) in cultured cardiomyocytes of neonatal rats were observed. Total protein contents were determined by Bradford method. The expression of c-fos protein was detected by Western blot. [Ca2+ ]i labeled with fluorescent probe Fluo-3/AM was measured under a laser scanning confocal microscope. The results revealed that as compared with control, the total protein contents were increased in cardiomyocytes treated with Ang Ⅱ (10-7 mol/L), which could be inhibited by CsA in a dose-dependent manner. It was found that Ang Ⅱ could increase the c-fos protein expression, which could be inhibited by CsA in a dose-dependent manner.Ang Ⅱ induced the [Ca2+ ]i elevation in cardiomyocytes. CsA did not influence the resting intracellular Ca2+ , but inhibited significantly the Ang Ⅱ-induced [Ca2+ ]i elevation. It was concluded that CsA can suppress the Ang Ⅱ-induced c-fos protein expression and [Ca2+ ]i elevation in single cardiomyocyte, which might play a role in the prevention of Ang Ⅱ -induced cardiomyocyte hypertrophy by CsA.     

Involvement of Ca^2＋-activated K^＋Channels in Receptor-Regulated Sperm Motility in Rats

Ion fluxes have been implicated in sperm functions.A pivotal role of Ca2 hasbeen observed in variousreproductive events including sperm capacitation,acrosomereaction[1～ 3] ,sperm activation[4～ 5] and sperm motility[6～ 8] .The importance of Ca2 in-f…     

ERK1/2信号通路在Ang-(1-7)抑制AngⅡ诱导的NRK52E转分化中的作用

目的探讨血管紧张素-(1-7)[Ang-(1-7)]对血管紧张素Ⅱ(AngⅡ)诱导的大鼠肾小管上皮细胞(NRK52E)表型转化及ERK1/2信号分子在转分化过程中的作用。方法体外培养NRK52E,经Ang-(1-7)和AngⅡ(终浓度均为10-6mol/L)干预24、48、72、96 h后,应用细胞免疫化学法检测E-cadherin、α-SMA的表达;干预72、96 h后,应用Western blot法检测P-ERK1/2表达水平的变化。结果 AngⅡ作用96 h后,E-cadherin的表达显著减弱(P<0.05),α-SMA的表达显著增强(P<0.05),P-ERK1/2表达显著增强(P<0.05);同时加入Ang-(1-7)后,与AngⅡ组比较,E-cadherin的表达显著增强(P<0.05),α-SMA的表达显著减弱(P<0.05),P-ERK1/2表达显著减弱(P<0.05)。结论 Ang-(1-7)能够抑制AngⅡ诱导的大鼠NRK52E表型转化,ERK1/2信号通路参与了肾小管上皮细胞转分化过程。     

同型半胱氨酸在大鼠血管平滑肌细胞上通过激活ERK1/2信号通路促进血管紧张素Ⅱ受体1表达

目的：在原代大鼠血管平滑肌细胞(VSMC)上观察同型半胱氨酸(Hcy)对血管紧张素II受体1(AT1R)的蛋白表达的影响。     

INHIBITORY EFFECT OF ANGIOTENSIN Ⅱ TYPE 1 RECEPTOR-ASSOCIATED PROTEIN ON VASCULAR SMOOTH MUSCLE CELL GROWTH AND NEOINTIMAL FORMATION

Objective To investigate the mechanism of a novel angiotensin Ⅱ type 1 receptor-associated protein （ATRAP） interfering with angiotensin Ⅱ type 1 （AT1） receptor-mediated vascular smooth muscle cell （VSMC） growth and neointimal formation. Methods VSMCs isolated from thoracic aorta of adult Sprague-Dawley （SD） rats were used in this study. ATRAP cDNA was subcloned into pcDNA3 vector and then transfected into VSMCs. DNA synthesis and extracellular signal-regulated kinase （ERK） and phospho-ERK expressions in VSMCs were assayed by measurement of ^3H thymidine incorporation and Western blotting, respectively. Morphological changes were observed in the balloon injured artery with or without transfection of ATRAP cDNA using 12-week-old male SD rats. Results ATRAP overexpression in VSMCs inhibited angiotensin Ⅱ （Ang Ⅱ）-induced ^3H thymidine incorporation 48 hours after Ang Ⅱ stimulation （ P 〈 0. 05 ）. In VSMC, Ang Ⅱ stimulation increased the phosphorylation of ERK, which reached the peak around 60 minutes. The activation of phospho-ERK was significantly decreased by ATRAP （ P 〈 0. 05 ）. Neointimal formation was markedly inhibited by ATRAP overexpression in injuried arteries. Conclusions The AT1 receptor-derived activation of ERK plays an essential role in Ang Ⅱ-induced VSMC growth. The growth inhibitory effects of ATRAP might be due to interfering with AT1 receptor-mediated activation of ERK in VSMC growth and neointimal formation.     

钙调神经磷酸酶信号通路在AngⅡ刺激的大鼠VSMCs增殖中的作用

目的:探讨钙调神经磷酸酶（calcineurin,CaN）依赖的信号通路在血管紧张素Ⅱ（Ang Ⅱ）刺激的大鼠血管平滑肌细胞（VSMCs）增殖中的作用。方法:采用组织贴块法,体外原代培养大鼠胸主动脉平滑肌细胞;Ang Ⅱ刺激培养的大鼠 VSMCs增殖;CaN特异性抑制剂环孢素 A（CsA）阻断Ang Ⅱ刺激的大鼠VSMCs中CaN依赖的信号转导通路;观察各因素对CaN活性、细胞增殖活度、增殖细胞核抗原（PCNA）表达水平的影响。结果:在一定范围内,Ang Ⅱ（10-8～10-6mol/L）以浓度依赖性方式增加VSMCs的CaN活性,同时细胞增殖活度（AMTT值）增高,与对照组相比,差异有统计学意义（P<0.01或P<0.001）。CsA（1 μmol/L）抑制CaN活性后,明显降低Ang Ⅱ（10-7mol/L）刺激的VSMCs增殖活度及核内PCNA的表达水平（OD值）,与Ang Ⅱ组比较,差异显著（P<0.001）。结论:CaN依赖的信号通路在Ang Ⅱ刺激的血管平滑肌细胞增殖中发挥重要作用。     

PP60c-Src在血管紧张素Ⅱ对大鼠血管平滑肌细胞信息转导中的作用

目的：通过观察c—Src在AngⅡ对大鼠血管平滑肌细胞(VSMC)丝裂原活化的蛋白激酶(MAPK)活性和c—fos蛋白表达的影响，以进一步了解AngⅡ促VSMC增殖的细胞内信息转导机制。方法：原代和传代培养SD大民主动脉VSMC，以脂质体包裹反义c—Src寡脱氧核夺酸(Oligodeoxynucleotides ODNs)转染培养的VSMC以抑制c—Src蛋白表达和激酶活性。以未转染的VSMC为对照，观察10^7mol/L AngⅡ刺激对转染的VSMC的MAPK活性和c—fos蛋白表达的影响。蛋白免疫沉淀和酶自身磷酸化率测定c—Src激酶活性；髓鞘碱性蛋白(MBP)底物磷酸化率测定MAPK放酶活性；Western blot免疫印迹法测定c—Src和c—fos蛋白表达情况。始果：转染不同浓度反义c—Src()DNs的VSMCc—Src蛋白含量至浓度依赖性降低，0.2μmol/L、0.5μmol/L、1．0μmol／L和2．0μmol/L分别为对照的68．2％、34．7％、30。3％和15．8％，经方差分析具有显著性意义(P＜0.01)。c—Src激酶活性也显著抑制；以AngⅡ刺激经转染反义c—Src DNs的VSMC，c—Src激酶活性增幅仅为对照组的8．7％；MAPK活性仅为对照的1．6％；c—fos蛋白表达的增幅为对照组的30．0％。结论：AngⅡ可诱导VSMC c—Src激活和细胞内信息转导，且AngⅡ引起的MAPK和c—fos的激活依赖于c—Src的激活，提示c—Src是AngⅡ促血管平滑细胞增殖的重要信息分于。     

宽叶缬草对动脉平滑肌细胞收缩及生长的影响

目的 :观察宽叶缬草 (VOL)对培养的血管平滑肌细胞 (VSMC)收缩及生长的影响。方法 :取 4～ 6周wistar大鼠胸主动脉中层平滑肌细胞进行原代及传代培养。观察VOL和L -NAME对VSMC收缩的影响 ,以及血管紧张素Ⅱ (AngⅡ )和不同浓度VOL时VSMC的3H -TdR及3H -Leucine参入变化。结果 :VOL可显著抑制AngⅡ引起的VSMC收缩 ,且此作用不受L -NAME影响。VOL呈剂量依赖性抑制VSMC的3H -TdR和3H -Leucine的参入。结论 :VOL有显著抑制AngⅡ引起的VSMC收缩和生长的作用     

Role of angiotensin Ⅱ and angiotensin Ⅱ receptors in vascular smooth muscle cell migration in vitro

ObjectiveTo determine the biotic effects of angiotensin Ⅱ (Ang Ⅱ) on the migration of rat smooth muscle cells (VSMCs) and investigate the mechanisms involved in the development of vascular injury. Methods VSMCs isolated from aortic media of Wistar rats and cultured by the modified explant method were adopted. In the presence and absence of Ang Ⅱ, the expression of Ang Ⅱ receptor (ATR) and reorganization of the actin cytoskeleton and focal adhesion of VSMCs were studied by an immunocytochemistry technique and fluorocytochemistry technique. Migration assays were performed with a modified Boyden’s chamber. The effects of AT(1)R antagonist (CV- 11974), AT2R antagonist (PD123319) on the aforementioned target were studied. Results VSMCs migration was stimulated by adding Ang Ⅱ. The dynamic reorganization of actin cytoskeleton and focal adhesions may be an important mechanism by which Ang Ⅱ facilitates VSMCs motility. The expression of AT(1)R in VSMCs could be upregulated initially after treatment with Ang Ⅱ, then decreased gradually. The expression of AT(1)R was downregulated by AT(1)R antagonists. The effect of Ang Ⅱ on VSMCs migration was mediated by AT(1)R, while AT2R had no significant effect. Conclusions The dynamic reorganization of focal adhesions and the actin cytoskeleton is required for Ang Ⅱ- induced VSMCs migration. This effect is mediated by AT(1)R.