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AICAR inhibits proliferation and induced S-phase arrest,and promotes apoptosis in CaSki cells

Acta Pharmacologica Sinica 2007年第12期28卷 1984-1990页

作者： Tong-ju GUAN~2,Feng-jin QIN~3.Jian-hai DU~4,Li Geng~3.You-yi ZHANG~(4,5),Min LI~(2,5)~2 institute of Pathology,The School of Basic medicine,Lanzhou university,Lanzhou 73.000,{3. ~3.Department of Gynaecology andObstetrics,Peking university third hospital,beijing 100083,{3. ~4 institute of vascular medicine,Peking university third hospital {13. laboratory of molecular cardiovascular sciences ministry of education,beijing 100083,{3. institute of Pathology The School of Basic Medicine Lanzhou University Lanzhou China Department of Gynaecology and Obstetrics Peking University Third Hospital Beijing China institute of vascular medicine Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education Beijing China

Aim:The aim of the present study was to determine the effect of 5-aminoimidazole-4-carboxamide-ribonucleoside （AICAR） on proliferation,cell cycle,and apoptosisin the human epithelial cervical cancer cell line CaSki ***:Cell countand 3.（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay were usedto determine cell proliferation and *** 3.258 staining was con-ducted to distinguish the apoptotic *** cycle and Annexin-V/propidiumiodide staining were analyzed by fluorescence-activated cell sorting （FACS）.AWestern blot assay was used to evaluate the expression of AKT （also known asprotein kinase B）,mammalian target of rapamycin （mTOR）,p53.and extracellularsignal-regulated kinase （ERK）.Results:AICAR （500 μmol/L） significantly inhibi-ted the proliferation of CaSki cells treated for 24,48,and 72 h as determined by *** cells at the G1and G2phases were dramatically decreased while cells atthe S phase were increased in response to AICAR treatment for 24,48,and 72 *** MTT assay showed less viable cells and Hoechst fluorescent staining showedmore apoptotic cells upon AICAR *** results of the Annexin-Vstaining demonstrated a time-dependent increase of apoptosis in cells treatedwith AICAR for 24,3.,and 48 ***,AICAR activated caspase-3.in atime-dependent *** was also found that AICAR inhibited the phosphory-lation of AKT and mTOR,which are important kinases regulating cell growth *** stimulation obviously increased the expression of the tumorsuppressor p53.and the phosphorylation of ***:AICAR inhibitedproliferation and induced S phase arrest and promoted apoptosis in CaSki cells,which might be mediated by the downregulation of the AKT/mTOR pathway andthe upregulation of the p53.ERK pathway.

关键词： AICAR proliferation apoptosis CaSki cells

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Crosstalk between signaling pathways of adrenoreceptors and signal transducers and activators of transcription 3.STAT3.in heart

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Acta Pharmacologica Sinica 2007年第2期28卷 153-161+163-165页

作者： Kai-zheng GONG Hui ZHANG Jian-hai DU You-yi ZHANG~2 institute of vascular medicine,Peking {3. third hospital,Key laboratory of molecular cardiovascular sciences,ministry of education beijing 100083,{13. institute of vascular medicine Peking University Third Hospital Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education Beijing China

Recently,there have been important advancements in our understanding of thesignaling mechanisms of adrenoreceptors(AR)and signal transducers and acti-vators of transcription 3.STAT3..While their crucial roles in the pathologicalprocesses of the heart are well established,accumulating evidence suggests thereis a complex pattern of crosstalk between these 2 signaling ***,the potential for crosstalk occurs at multiple levels in each signaling cascade andinvolves receptor transactivation,G proteins,small GTPases,cyclic adenosine 3.,5’-monophosphate/protein kinase A,protein kinase C,scaffold/adaptor proteins,protein tyrosine kinases,and mitogen-activated protein *** addition,post-translational modification(eg acetylation)of STAT3.may provide a link betweenSTAT3.and AR *** particular,crosstalk between these 2 systems in theheart would appear to be dependent upon the species/tissue studied,develop-mental stage,and eliciting *** at least partly accounts for the epige-netic effects on biological function that is mediated by the 2 signaling *** of these mechanisms will provide new targets in the development ofnovel clinical strategies for heart disorders.

关键词： receptors adrenergic signal transduction STAT3.transcription factor heart

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Internalization and distribution of three α1-adrenoceptor subtypes in HEK293. cells before and after agonist stimulation

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Acta Pharmacologica Sinica 2007年第3期28卷 359-366页

作者： Shu-yi WANG~2 Yao SONG~2 Ming XU~2 Qi-hua HE~3.Qi-de HAN~2 You-yi ZHANG~(2,4)2 institute of vascular medicine,Peking {3. third hospital and Key laboratory of molecular cardiovascular sciences,ministry of education,beijing 100083,{13. 2 Center of Medical Analysis,Peking {3.,Health Science Center,beijing 100083,{13. institute of vascular medicine Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education Beijing China Center of Medical Analysis Peking University Health Science Center Beijing China

Aim:To examine the subcellular distribution of the 3.α1-adrenoceptor（α1-AR）subtypes and their internalization and trafficking upon agonist stimulation in hu-man embryonic kidney 293. ***:Confocal real-time imaging,enzymelinked immunosorbent assay（ELISA）and whole cell[3.]-prazosin binding assaywere applied to detect the distribution and localization of the 3.α1B-AR ***:α1A-AR was found both on the cell surface and in the cytoplasm;α1B-AR,however,was predominantly detected on the cell surface,while α1D-AR wasdetected mainly in the intracellular *** stimulation withphenylephrine,localization changes were detected by confocal microscopy forα1A-and α1B-AR, but the localization of α1D-AR were *** promoted a more rapid internalization of α1B-AR than α1A-AR.α1D-ARinternalization was detected only by *** cell[3.]-prazosin bindingassay showed that α1A-AR functional receptors were detected both on the cellsurface and in the cytoplasm;α1B-AR,however,were detected predominantly onthe cell surface,while α1D-AR were detected mainly in intracellular *** stimulation promoted internalization of α1A-and α***:Phenylephrine stimulation induced changes in the localization of the3.1-AR.

关键词： adrenoceptor agonist stimulation confocal image enzyme-linked immunosorbent assay [~3.]-prazosin binding assay internalization subcellular distribution subtype

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Real-time detection of α1A-AR movement stimulated by phenylephrine in single living cells

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Acta Pharmacologica Sinica 2007年第6期28卷 796-802页

作者： Ning XU~(2,5),Zhang-yi LIANG~(3.5),Ming XU~2,Ying-hua GUAN~3.Qi-hua HE~4,Qi-de HAN~2,Xin-sheng ZHAO~(3.6),You-yi ZHANG~(2,6)~2 institute of vascular medicine,Peking {3. third hospital,and Key laboratory of molecular cardiovascular sciences,ministry ofeducation,beijing 100083,{13. ~3.beijing National laboratory for molecular sciences,State Key laboratory for Structural Chemistry ofUnstable and Stable Species,and Department of Chemical Biology,College of Chemistry and molecular Engineering,Peking {3.,beijing 100871,{13. ~4 Center of Medical Analysis,Peking {3.,Health Science Center,beijing 100083,{13. institute of vascular medicine Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education Beijing China beijing National laboratory for molecular {3. State Key Laboratory for Structural Chemistry of Unstable and Stable Species Department of Chemical Biology College of Chemistry and Molecular Engineering Peking University Beijing China Center of Medical Analysis Peking University Health Science Center Beijing China

Aim:To investigate the movement of α1A-adrenergic receptors(α1A-AR) stimu-lated by agonist,phenylephrine （PE）,and the dynamics of receptor movement inreal time in single living cells with millisecond ***:We labeledα1A-AR using the monoclonal,anti-FLAG （a kind of tag） antibody and Cy3.conju-gated goat anti-mouse IgG and recorded the trajectory of their transport processin living HEK293. cells stimulated by agonist,PE,and then analyzed their dy-namic ***:The specific detection of α1A-AR on the surface ofliving HEK293.-α1Acells was achieved.α1A-AR internalize under the stimulationof *** the cells were stimulated with PE for 20 min,apparent colocalizationwas found between α1A-AR and *** 40 min stimulation of PE,trajecto-ries of approximate linear motion in HEK293.-α1Acells were recorded,and theirvelocity was ***:The specific labeling method on the livingcell surface provides a convenient means of real-time detection of the behavior ofsurface *** this method we were able to specifically detect α1A-AR andrecord the behavior of individual particles of receptors with 50 ms exposure time inreal time in single living cells.

关键词： adrenergic receptor trajectory living cell

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Distinct actions of intermittent and sustained β-adrenoceptor stimulation on cardiac remodeling

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Science china(Life sciences) 2011年第6期54卷 493-501页

作者： MA XiaoWei1,3.SONG Yao1,3.CHEN Chao1,3.FU YongNan1,3.SHEN Qiang2,3.LI ZiJian1,3.& ZHANG YouYi1,3.1institute of vascular medicine,Peking {3. third hospital,beijing 100191,china 2institute of cardiovascular Science,Peking {3.,beijing 100191,china 3.ey {13. of molecular cardiovascular sciences,ministry of education,beijing 100191,china institute of vascular medicine Peking University Third Hospital Beijing China Key laboratory of molecular cardiovascular {3. Ministry of Education Beijing China institute of cardiovascular Science Peking University Beijing China

Heart disease is associated with increased sympathetic nerve activity and elevated levels of circulating catecholamines,resulting in chronic stimulation of the β-adrenergic receptors (β-AR) and consequent pathological cardiac ***,chronic administration of the β-AR agonist isoproterenol (ISO) has been most commonly used to model β-AR-induced cardiac ***,it remains unclear whether β-AR-mediated cardiac remodeling and dysfunction differs between sustained versus pulsatile (intermittent) exposure to a β-***,we compare the effects of intermittent versus sustained administration of ISO on cardiac remodeling and function in *** were administered 5 mg (kg d)-1 ISO for 2 weeks either by daily subcutaneous injection,or continuous infusion via an implanted osmotic *** function and remodeling were determined by echocardiography,micromanometry and ***,Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were utilized to define the proteins and genes *** sustained and intermittent administration of ISO resulted in a similar degree of cardiac hypertrophy (16% and 19%,respectively).However,mice receiving ISO by daily injection developed more severe ventricular systolic and diastolic dysfunction and myocardial fibrosis compared with mice receiving ISO via the osmotic *** disparity in results between the delivery methods is suggested to be due,at least in part,to increased expression of fibrogenic factors,including connective tissue growth factor (CTGF) and NADPH oxidase (NOX4),in mice receiving intermittent application of *** summary,compared with sustained exposure to a β-AR agonist,intermittent β-AR stimulation leads to more severe cardiac dysfunction and *** findings not only further our understanding of β-AR function in the setting of cardiac pathophysiology,but also highlight that significant differences can result dependent upon the mod

关键词： β-adrenoceptor reactive oxygen species NADPH oxidase cardiac hypertrophy cardiac fibrosis

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Intermittent hypobaric hypoxia protects the heart against ischemia-reperfusion injury through activation of α1-adrenoceptors

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中国病理生理杂志 2012年第11期 2020-2021页

作者： GAO Ling1,CHEN Le1,LU Zhi-zhen2,ZHANG You-yi 2,ZHANG Cai-mei1,CHEN Yi-xiong1,YANG Huang-tian1(1Key laboratory of Stem Cell Biology and laboratory of molecular Cardiology,{3. of Health sciences,Shanghai {3.s for Biological sciences,Chinese Academy of sciences,Shanghai 200025,china 2{3. of vascular medicine,Peking university {13. hospital & Key laboratory of molecular cardiovascular sciences ministry of education,beijing 100191,china)

AIM:Cumulated evidence have shown that intermittent hypobaric hypoxia(IHH)can protect the heart against ischemia-reper-fusion(I/R)injury by improving major manifestations such as infarct size,post-ischemic contractile... 详细信息

关键词： Intermittent hypobaric hypoxia protects the heart against ischemia-reperfusion injury through activation of adrenoceptors AIM MMP

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α1A-adrenergic receptor mediated pressor response to phenylephrine in anesthetized rat

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Science china Life sciences 2004年第1期47卷 59-65页

作者： XU Qi, ZHU Weizhong, LU Zhizhen, ZHANG Youyi & HAN Qide institute of vascular medicine, Peking {3. third hospital Key laboratory of molecular Cardiology, ministry of education, beijing 100083, china institute of vascular medicine Peking University Third Hospital Key laboratory of molecular Cardiology Ministry of Education Beijing 100083 China

To determine which subtype of α1-adrenergic receptors plays a role in the regulation of blood pressure, with α1A-adrenergic receptor-mediated vasoconstriction in perfused hindlimb as a control, we compared the inhibitory effects of various ai-adrenergic receptor selective antagonists on the vasopressure responses to phenylephrine between the mean arterial pressure and hindlimb perfusion pressure in anesthetized rats. In Normotensive Wistar rats, the results showed that the inhibitory effects (dose ratios of ED50, Dr) of α1-adrenoceptor selective antagonist (prazosin, Dr 13.5±3.6 vs.15.1±4.3. n = 11), α1A-adrenoceptor selective antagonist (5-methyl-urapidil, Dr 2.4±0.9 vs. 3.7±2.3. n = 12; RS-17053. Dr 3.2±1.6 vs. 4.4±3.3. n =12) and α1D-adrenoceptor selective antagonist (BMY73.8, Dr 1.9±0.9 vs. 2.2±0.8, n = 8) on phenylephrine- induced increases of perfusion pressure in the autoperfused femoral beds were the same as that in the mean arterial blood pressure in normotensive Wistar rats. The i

关键词： α1-adrenergic receptor blood pressure antagonist spontaneous hypertensive rats.

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Intermolecular Failure of L-type Ca2+ Channel and Ryanodine Receptor Signaling in Hypertrophy

Intermolecular Failure of L-type Ca2+ Channel and Ryanodine ...

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中国生理学会张锡钧基金会第十届全国青年优秀生理学学术论文交流会

作者： Ming Xu~#,Peng Zhou~#,Shi-Ming Xu~#,Yin Liu,Xinheng Feng,Shu-Hua Bai,Yan Bai, Xue-Mei Hao,Qide Han,Youyi Zhang~*,Shi-Qiang Wang~* (State Key Lab of Biomembrane and Membrane Biotechnology,ministry of education Key Lab of molecular {3. sciences and institute of vascular medicine, third hospital,College of Life sciences,Peking university,beijing,{13.)

High blood pressure induces hypertrophy,a thickening of the cardiac muscle that eventually leads to heart failure,a leading cause of morbidity and *** contractile power of the heart depends in part on signaling between calcium channels on the cell membrane(L-type Ca charnels) and calcium release channels on a specialized calcium-regulating organelle called the sarcoplasmic *** signaling process is defective in heart *** have found that the signaling

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Junctophilin-2表达下调介导心肌细胞兴奋—收缩耦联结构与功能的重塑

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中国分子心脏病学杂志 2013年第3期13卷 543-547页

作者：马元良黎荣昌吴昊迪王世强徐明北京大学第三医院心内科血管医学研究所卫生部心血管分子生物学与调节肽重点实室分子心血管教育部重点实验室北京市100191 北京大学生命科学院生物膜与膜生物工程国家重点实验室北京市100871

目的阐明横管(T-tubule,TT)与肌质网(sarcoplasmic reticulum,SR)耦联关键蛋白junctophilin-2(JP2)的表达下调对心肌细胞二联体超微结构以及兴奋-收缩耦联功能的影响。方法通过构建JP2特异性shRNA腺病毒敲减成年大鼠心肌细胞中JP2的表达... 详细信息

目的阐明横管(T-tubule,TT)与肌质网(sarcoplasmic reticulum,SR)耦联关键蛋白junctophilin-2(JP2)的表达下调对心肌细胞二联体超微结构以及兴奋-收缩耦联功能的影响。方法通过构建JP2特异性shRNA腺病毒敲减成年大鼠心肌细胞中JP2的表达后,采用透射电子显微镜观测心肌细胞横管和肌质网二联体结构的形态变化,并进一步采用全细胞膜片钳结合激光共聚焦钙成像技术检测心肌细胞兴奋收缩联联功能的变化。结果通过腺病毒敲减心肌细胞JP2表达后,心肌细胞内总TT数目、TT与SR耦联的二联体结构数目以及单个耦联子内耦联SR的TT长度占TT周长的百分比均显著降低;与此同时,JP2表达下降后心肌细胞钙瞬变幅度降低,收缩功能减弱,表现出与心力衰竭类似的表型。结论心肌细胞JP2表达下降引起心肌细胞兴奋-收缩耦联结构与功能的损伤,为心力衰竭病理情况下心肌细胞结构与功能的调控机制提供了直接有力的实验证据。

关键词： junctophilin-2 心肌细胞二联体结构兴奋-收缩耦联

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