S100A1基因转导联合骨髓间充质干细胞移植治疗慢性心力衰竭大鼠的实验研究
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摘要
背景与目的慢性心力衰竭(chronic heart failure,CHF)已成为心血管疾病死亡的重要原因。心力衰竭的本质是心肌细胞数量的减少和分子机制异常。针对心肌细胞数量减少,移植能具有多向分化和自我增殖潜能的骨髓间充质干细胞(Mesenchymal stemcells,MSCs)虽能改善CHF患者心脏功能,但是并不能解决心力衰竭的分子机制异常。现研究表明,CHF分子机制异常的关键是Ca~(2+)转运异常,表现为Ca~(2+)的摄入、释放、转运障碍。人们曾针对Ca~(2+)摄入障碍增加肌浆网钙ATP酶(sarcoplasmic reticulumCa2+-ATPase,SERCA2a)表达,发现虽能增强心脏收缩功能,但会增加心肌梗死急性期死亡率、心律失常发生率。钙结合蛋白S100A1(calcium-binding protein S100A1)为调控SERCA2a的上游基因,通过对肌浆网钙SERCA2a、兰尼碱(ryanodine receptors 2,RyR2)受体的调控,调节心肌细胞Ca~(2+)的释放、摄取、转运。CHF时S100A1蛋白表达下降,增加S100A1蛋白表达既能改善心脏功能,阻止心力衰竭的进展,又没有增加心律失常等副作用。MSCs与S100A1联合移植是否能达到既修复心肌细胞又调控衰竭心肌分子机制而达到改善心功能的相加效应,目前国内外均无报道。本研究拟通过观察对照组、MSCs移植组、S100A1转导组、MSCs与S100A1联合移植组心功能效应,探讨S100A1基因转导、S100A1基因转导联合MSCs移植在心力衰竭大鼠中的心功能效应。方法取6—8周龄雄性SD(Sprague-Dawley)大鼠胫骨、股骨,收集骨髓腔冲洗悬液直接培养,经多次换液、传代,免疫组化法检测CD34、CD44表达情况。以超顺磁氧化铁和P3代MSCs共同孵育培养24h,普鲁士蓝染色评价标记效率,台盼蓝染色检验标记后细胞的活性。取8—10周龄SD大鼠50只,随机分至实验组(n=40)和假手术组(n=10)。以氯胺酮、速眠新Ⅱ大腿内侧肌肉注射复合麻醉,开胸后以置液氮内预冷10分钟的金属棒间断冷冻左心室游离壁3次,持续时间分别为15s,15s,10s。术后行心电图、超声心动图检查,取心脏组织行氯化三苯四氮唑(triphenyltetrazolium chloride,TTC)、苏木素-依红(hematoxylin-eosin,H&E)染色,观察心肌梗死面积、心功能变化,观察术中、术后大鼠存活率。在冷冻法建立的稳定慢性心力衰竭大鼠模型上,开胸后按分组分别将生理盐水、MSCs悬液、S100A1-AAV1/2病毒、S100A1-AAV1/2+MSCs混合物注射至大鼠心肌梗死灶周围。4周后用超声诊断仪测量心功能,取心脏组织行普鲁士蓝染色示踪移植的MSCs。结果全血贴壁法培养的大鼠MSCs贴壁、呈纤维细胞样生长,CD44阳性细胞比率为96%,不表达CD34。经SPIO标记后,普鲁士蓝染色标记效率为90%,标记后98%的细胞保持活性。冷冻术后4周实验组40只大鼠中35只两个或两个以上相邻导联Q波形成且＞1mv或V1、V3、V5导联R波之和＜10mv,超声心动图左室射血分数(left ventricular ejectionfraction,LVEF)46.55±6.80%,左室短轴缩短率(fractional shortening,FS)20.61±3.61%,TTC染色证实实验组左室梗死面积35.62±3.74%,HE染色示梗死区主要为疤痕组织,边界清晰。假手术组未见心肌梗死。实验组术后4周存活率达到87.5%。S100A1-AAV1/2+MSCs组、MSCs组心肌梗死灶边缘见普鲁士蓝染色阳性细胞,对照组未见任何阳性细胞。S100A1-AAV1/2+MSCs组、S100A1-AAV1/2组、MSCs组左室射血分数较治疗前分别增加15.41±6.75%(P＜0.01)、7.85±2.96%(P＜0.01)、5.63±2.60%(P＜0.01),左室短轴缩短率较治疗前分别增高6.17±3.46%(P＜0.01)、3.02±1.25%(P＜0.01)、1.58±0.59%;左室射血分数、左室短轴缩短率增加值S100A1-AAV1/2+MSCs组与S100A1-AAV1/2组、MSCs相比有统计学差异(P＜0.05)。S100A1-AAV1/2+MSCs组、S100A1-AAV1/2组左室舒张末直径(left ventricular enddiastolic diameter,LVEDD)分别较治疗前减小1.87±0.64mm(P＜0.01)、1.00±0.62mm(P＜0.05);左室舒张末直径减少值S100A1-AAV1/2+MSCs组与S100A1-AAV1/2组、MSCs相比有统计学差异(P＜0.01)。对照组左室射血分数、左室短轴缩短率较治疗前分别降低8.44±3.74%(P＜0.01)、4.64±1.37%(P＜0.01),左室舒张末直径增大0.9±0.14mm(P＜0.01)。结论全血贴壁法能成功培养大鼠MSCs,SPIO可以安全、有效地标记MSCs,并能在体内示踪。采用金属棒冷冻法可成功建立大鼠急性心肌梗死后慢性心力衰竭模型,有效地提高术后大鼠存活率。以腺相关病毒为载体S100A1基因转导可以明显增强心力衰竭大鼠心脏功能,与MSCs联合可以进一步增加心力衰竭大鼠心脏功能,S100A1的过表达可以阻止心力衰竭的进展。
Background and Objectives Chronic heart failure is an important cause of coronary heart disease death.Myocardiocyte reduction and molecular defect are crucial in chronic heart failure(CHF).Mesenchymal stem cells(MSCs) has the potential to regenerate and mutidifferentiate into myocardiocyte like cells.Evidence suggests that transplanting MSCs into the failing heart can prevent deleterious remodeling and improve recovery.But the stem cells transplantation does not solve the molecular defect.Ca~(2+) carrying defect is an important aspect of molecular defect.Increasing sarcoplasmic reticulum Ca~(2+)-ATPase(SERCA2a) expression can improve the systolic function but impaired survival in response to ischemic cardiac injury under clinical conditions.S100A1 protin is a Ca~(2+)-depending inotropic factor in heart,interacts both with the SR(sarcoplasmic reticulum,SR) Ca~(2+) release channel(RyR2) and the SERCA2a in a Ca~(2+) dependent manner,resulting in enhanced Ca~(2+) induced SR Ca~(2+) release and SR Ca~(2+) reuptake,respectively.In addition,S100A1 can decrease the diastolic SR Ca~(2+) leak through attenuated Ca~(2+) spark frequency,suggesting a biphasic Ca~(2+) dependent modulation of the RyR2 through interaction with S100A1.Evidence suggest that S100A1 protin expression decreased in failuring heart and increasing S100A1 protin expression in the failing heart can improve cardic function and inhibite left ventricular remodeling and does not increase arrhythymal rate.Wether S100A1 gene combing MSCs transplantation for CHF can improve the cardic function more,there is no report.In order to inquire this question,we study the cardic fucion in rat CHF model by echocardiography in the control group,MSCs group,rAAV1/2-S100A1 group,MSCs and rAAV1/2-S100A1 group.Methods Rat bone marrow-derived MSCs were harvested from 6-to 8-week-old(150-80g) male Sprague-Dawley rats.After the bone marrow plugs were flushed out from tibial and femoral bones,the whole marrow samples were collected and cultured with Dulbecco's modified Eagle's medium-low glucose(L-DMEM).After identification the specific surface antigens CD34 and CD44 by immunohistochemistry,the third passage MSCs were incubated with superparamagnetic iron oxide(SPIO) for 24hours.The labeling efficiency was tested through Prussian blue staining,the cells viability was tested through Trypan blue rejection method.After that,fifty SD rats were randomized into experiment and sham-operated groups.After being anesthetized and artificially ventilated,a thoracotomy was performed through the fourth left intercostal space.Cryoinfarction was produced by applying steal cryoprobes of 6mm in diameter which were immersed in liquid nitrogen for 10 min to the anterior left ventricular free wall for 3 times.The during time was 15s,15s,10s respectively.By electrocardiogram(ECG),echocardiography, hematoxylin-eosin(H & E) staining and triphenyltetrazolium chloride(TTC) staining to evaluation the infarct size and heart function.The survival rate during operation and in 4 weeks after operation was observed.Then the CHF rats were devided four groups:control group,MSCs group, rAAV1/2-S100A1 group,MSCs and rAAV1/2-S100A1 group.MSCs were labeled with superparamagnetic iron oxide(SPIO).After performed thoracotomy,0.9%sodium chloride solution, mesenchymal stem cells,recombinant adeno-associated viral 1/2 vector carrying S100A1 gene,the mixtures of mesenchymal stem cells and recombinant adeno-associated viral 1/2 vector carrying S100A1 gene were respectively injected into intramyocardial of heart failure rats in different groups.Then at 4 weeks after treament,we evaluate the cardiac function by echocardiography, identify the transplanted mesenchymal stem cells through Prussian blue staining.Results Most cells were spindle-like in shape and adhered to the plastic tissue culture dishes. Immunohistochemistry showed that 96%cells express CD44 but no cells express CD34. After incubated with SPIO(superparamagnetic iron oxide),the labeling efficiency and the cells viability is 90%,98%,respectively.Infarct evidences were found in 35 of 40 experiment rats which survive over 4 weeks after operation.The infarct size was 35.62±3.74%in all experiment rats.The left ventricular ejection fraction(LVEF) was 46.55±6.80%,the fractional shortening(FS) was 20.61±3.61%.Transmural cryoinfarcted and distinct infarct boundary were observed by the H & E staining and triphenyltetrazolium chloride staining.No infarction was observed in sham-operation group.The heart function decreased significantly in the experiment rats than the sham-operated group(P＜0.01). The survival rate after 4 weeks was 87.5%.4 weeks afert treatment,the MSCs group,MSCs and rAAV1/2-S100A1 group were found Prussian blue staining positive cells;no positive cells were found in the control group.In the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group,MSCs group,the left ventricular ejection fraction increased 15.41±6.75%(P＜0.01),7.85±2.96%(P＜0.01),5.63±2.60%(P＜0.01) than before treatmeat respectively.And the fractional shortening in the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group,MSCs group increasrd 6.17±3.46%(P＜0.01),3.02±1.25%(P＜0.01),1.58±0.59%than before treatment respectively.The left ventricular end diastolic diameter(LVEDD) in the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group decreased 1.87±0.64mm(P＜0.01),1.00±0.62mm(P＜0.05) than before treatment respectively.In contrast to rAAV1/2-S100A1 group or MSCs group,the cardic function improved remarkably in MSCs and rAAV1/2-S100A1 group(P＜0.05).In the control group, left ventricular ejection fraction and fractional shortening decreased 8.44±3.74%(P＜0.01),4.64±1.37%(P＜0.01) respectively.The left ventricular end diastolic diameter increased 0.9±0.14mm(P＜0.01) than before treatment too.Conclusions By adherence the whole marrow samples can culture MSCs,and SPIO can lable MSCs efficiently and safely.Using ultralow temperature cryoinjury may be an ideal method to establish the rat model of acute myocardial infarction.This method can decrease mortality in making a successful rat chronic heart failure model.Increased calcium-binding protein S100A1 expression in chronic heart failure rats by transfer recombinant adeno-associated viral1/2 vector containing calcium-binding protein S100A1 gene can improve cardiac function. Transplant the mixtures of mesenchymal stem cells and rAAV1/2-S100A1 can get better effects than giving rAAV1/2-S100A1 or MSCs only.Increased calcium-binding protein S100A1 expression can inhibit the progress of heart failure.
Background and Objectives Chronic heart failure is an important cause of coronary heart disease death.Myocardiocyte reduction and molecular defect are crucial in chronic heart failure(CHF).Mesenchymal stem cells(MSCs) has the potential to regenerate and mutidifferentiate into myocardiocyte like cells.Evidence suggests that transplanting MSCs into the failing heart can prevent deleterious remodeling and improve recovery.But the stem cells transplantation does not solve the molecular defect.Ca~(2+) carrying defect is an important aspect of molecular defect.Increasing sarcoplasmic reticulum Ca~(2+)-ATPase(SERCA2a) expression can improve the systolic function but impaired survival in response to ischemic cardiac injury under clinical conditions.S100A1 protin is a Ca~(2+)-depending inotropic factor in heart,interacts both with the SR(sarcoplasmic reticulum,SR) Ca~(2+) release channel(RyR2) and the SERCA2a in a Ca~(2+) dependent manner,resulting in enhanced Ca~(2+) induced SR Ca~(2+) release and SR Ca~(2+) reuptake,respectively.In addition,S100A1 can decrease the diastolic SR Ca~(2+) leak through attenuated Ca~(2+) spark frequency,suggesting a biphasic Ca~(2+) dependent modulation of the RyR2 through interaction with S100A1.Evidence suggest that S100A1 protin expression decreased in failuring heart and increasing S100A1 protin expression in the failing heart can improve cardic function and inhibite left ventricular remodeling and does not increase arrhythymal rate.Wether S100A1 gene combing MSCs transplantation for CHF can improve the cardic function more,there is no report.In order to inquire this question,we study the cardic fucion in rat CHF model by echocardiography in the control group,MSCs group,rAAV1/2-S100A1 group,MSCs and rAAV1/2-S100A1 group.Methods Rat bone marrow-derived MSCs were harvested from 6-to 8-week-old(150-80g) male Sprague-Dawley rats.After the bone marrow plugs were flushed out from tibial and femoral bones,the whole marrow samples were collected and cultured with Dulbecco's modified Eagle's medium-low glucose(L-DMEM).After identification the specific surface antigens CD34 and CD44 by immunohistochemistry,the third passage MSCs were incubated with superparamagnetic iron oxide(SPIO) for 24hours.The labeling efficiency was tested through Prussian blue staining,the cells viability was tested through Trypan blue rejection method.After that,fifty SD rats were randomized into experiment and sham-operated groups.After being anesthetized and artificially ventilated,a thoracotomy was performed through the fourth left intercostal space.Cryoinfarction was produced by applying steal cryoprobes of 6mm in diameter which were immersed in liquid nitrogen for 10 min to the anterior left ventricular free wall for 3 times.The during time was 15s,15s,10s respectively.By electrocardiogram(ECG),echocardiography, hematoxylin-eosin(H & E) staining and triphenyltetrazolium chloride(TTC) staining to evaluation the infarct size and heart function.The survival rate during operation and in 4 weeks after operation was observed.Then the CHF rats were devided four groups:control group,MSCs group, rAAV1/2-S100A1 group,MSCs and rAAV1/2-S100A1 group.MSCs were labeled with superparamagnetic iron oxide(SPIO).After performed thoracotomy,0.9%sodium chloride solution, mesenchymal stem cells,recombinant adeno-associated viral 1/2 vector carrying S100A1 gene,the mixtures of mesenchymal stem cells and recombinant adeno-associated viral 1/2 vector carrying S100A1 gene were respectively injected into intramyocardial of heart failure rats in different groups.Then at 4 weeks after treament,we evaluate the cardiac function by echocardiography, identify the transplanted mesenchymal stem cells through Prussian blue staining.Results Most cells were spindle-like in shape and adhered to the plastic tissue culture dishes. Immunohistochemistry showed that 96%cells express CD44 but no cells express CD34. After incubated with SPIO(superparamagnetic iron oxide),the labeling efficiency and the cells viability is 90%,98%,respectively.Infarct evidences were found in 35 of 40 experiment rats which survive over 4 weeks after operation.The infarct size was 35.62±3.74%in all experiment rats.The left ventricular ejection fraction(LVEF) was 46.55±6.80%,the fractional shortening(FS) was 20.61±3.61%.Transmural cryoinfarcted and distinct infarct boundary were observed by the H & E staining and triphenyltetrazolium chloride staining.No infarction was observed in sham-operation group.The heart function decreased significantly in the experiment rats than the sham-operated group(P＜0.01). The survival rate after 4 weeks was 87.5%.4 weeks afert treatment,the MSCs group,MSCs and rAAV1/2-S100A1 group were found Prussian blue staining positive cells;no positive cells were found in the control group.In the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group,MSCs group,the left ventricular ejection fraction increased 15.41±6.75%(P＜0.01),7.85±2.96%(P＜0.01),5.63±2.60%(P＜0.01) than before treatmeat respectively.And the fractional shortening in the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group,MSCs group increasrd 6.17±3.46%(P＜0.01),3.02±1.25%(P＜0.01),1.58±0.59%than before treatment respectively.The left ventricular end diastolic diameter(LVEDD) in the MSCs and rAAV1/2-S100A1 group,rAAV1/2-S100A1 group decreased 1.87±0.64mm(P＜0.01),1.00±0.62mm(P＜0.05) than before treatment respectively.In contrast to rAAV1/2-S100A1 group or MSCs group,the cardic function improved remarkably in MSCs and rAAV1/2-S100A1 group(P＜0.05).In the control group, left ventricular ejection fraction and fractional shortening decreased 8.44±3.74%(P＜0.01),4.64±1.37%(P＜0.01) respectively.The left ventricular end diastolic diameter increased 0.9±0.14mm(P＜0.01) than before treatment too.Conclusions By adherence the whole marrow samples can culture MSCs,and SPIO can lable MSCs efficiently and safely.Using ultralow temperature cryoinjury may be an ideal method to establish the rat model of acute myocardial infarction.This method can decrease mortality in making a successful rat chronic heart failure model.Increased calcium-binding protein S100A1 expression in chronic heart failure rats by transfer recombinant adeno-associated viral1/2 vector containing calcium-binding protein S100A1 gene can improve cardiac function. Transplant the mixtures of mesenchymal stem cells and rAAV1/2-S100A1 can get better effects than giving rAAV1/2-S100A1 or MSCs only.Increased calcium-binding protein S100A1 expression can inhibit the progress of heart failure.
引文
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[14]Most P,Pleger ST,Volkers M,et al.Cardiac adenoviral S100A1 gene delivery rescues failing myocardium[J].J Clin Invest.2004;114(11):1550-1563.
[15]Pleger ST,Most P,Boucher M,et al.Stable Myocardial-specific AAV6-S100A1 gene therapy results in chronic functional heart failure rescue[J].Circulation.2007;115(19):2506-2515
[16]Kraitchman DL,Heldman AW,Atalar E,et al.In Vivo Magnetic Resonance Imaging of Mesenchymal Stem Cells in Myocardial Infarction[J].Circulation,2003,107(18):2290-2293.
[17]Arbab AS,Bashaw LA,Miller BR,et al.Characterization of Biophysical and Metabolic Properties of Cells Labeled with Superparamagnetic Iron Oxide Nanoparticles and Transfection Agent for Cellular MR Imaging[J].Radiology,2003,229(3):838-846.
[1]Pleger ST,Most P,Boucher M,Soltys S,Chuprun JK,Pleger W,Gao E,Dasgupta A,Rengo G,Remppis A,Katus HA,Eckhart AD,Rabinowitz JE,Koch WJ.Stable Myocardial-specific AAV6-S100A1 gene therapy results in chronic functional heart failure rescue.Circulation.2007;115:2506-2515.
[2]Most P,Pleger ST,Volkers M,Heidt B,Boerries M,Weichenhan D,Loffier E,Janssen PM,Eckhart AD,Martini J,Williams ML,Katus HA,Remppis A,Koch WJ.Cardiac adenoviral S100A1 gene delivery rescues failing myocardium.J Clin Invest.2004;114:1550-1563.
[3]Donato R.Intracellular and extracellular roles of S100 proteins.Microsco Res Tech.2003;60:540-551.
[4]Kiewitz R,Lyons GE,Schafer BW,Heizmann CW.Transcriptional regulation of S100A1 and expression during mouse heart development.Biochimica et Biophysica Acta.2000; 1498:207-219.
[5]Du XJ,Cole TJ,Tenis N,Gao XM, Kontgen F,Kemp BE,Heierhorst J. Impaired cardiac contractility response to hemodynamic stress in S100A1-deficient mice.Mol Cell Biol.2002; 22;2821-2829.
[6]Volkers M.Loughrey CM,Macquaide N,Reppis A,Degeorge B,Wegner F,Friedrich O,Fink RH,Koch WJ,Smith GL,Most P.S100A1 decreases calcium spark frequency and alters their spatial characteristics in permeabilized adult ventricular cardiomyocytes.Cell Calcium.2007; 41:135-143.
[7]Remppis A,Greten T,Schafer BW,Hunziker P,Erne P,Katus HA,Heizmann CW. Altered expression of the Ca(2+)-binding protein S100A1 in human cardiomyopathy. Biochim BiophysActa.1996;1313:253-257.
[8]Most P,Seifert H,Gao E,Funakoshi H,Volkers M,Heierhorst J,Remppis A,Pleger ST,DeGeorge BR Jr,Eckhart AD,Feldman AM.Koch WJ.Cardiac S100A1 protein levels determine contractile performance and propensity towards heart failure after myocardial infarction. Circulation. 2006; 114:1258 -1268.
[9]Hasenfuss,G.Alterations of calcium-regulatory proteins in heart failure. Cardiovasc Res. 1998; 37:279-289.
[10]Kiewitz R,Acklin C, Schafer BW,Maco B,Uhrik B,Wuytack F,Erae P, Heizmann CW.Ca2+-dependent interaction of S100A1 with the sarcoplasmic reticulum Ca2+-ATPase2a and phospholamban in the human heart. Biochem Biophys Res Commun. 2003;306:550-557.
[11]Lokuta AJ,Maertz NA,Meethal SV,Potter KT,Kamp TJ,Valdivia HH,Haworth RA. Increased nitration of sarcoplasmic reticulum Ca2 + ATPase in human heart failure.Circulation.2005; 111:988 -995.
[12]Milting H,Lukas N,Klauke B,Korfer R,Perrot A ,Osterziel KJ, Vogt J, Peters S, Thieleczek R,Varsanyi M.Composite polymorphisms in the ryanodine receptor 2 gene associated with arrhythmogenic right ventricular cardiomyopathy.Cardiovasc Res. 2006;71:496-505.
[13]Pleger ST,Remppis A,Heidt B,Volkers M,Chaprun JK, Kuhn M,Zhou RH,Gao E, Szabo G, Weichenhan D,Muller OJ,Eckhart AD,Katus HA,Koch WJ,Most P.S100A1 gene therapy preserves in vivo cardiac function after myocardial infarction.Mol Ther.2005;12:1120-1129.
[14]Most P,Boerries M, Eicher C,Schweda C,Ehlermann P,Pleger ST,Loffler E,Koch WJ,Katus HA,Schoenenberger CA,Remppis A. Extracellular S100A1 protein inhibits apoptosis in ventricular cardiomyocytes via activation of the extracellular-regulated kinase (ERK1/2) pathway. J Biol Chem.2003;278:48404-48412.
[15]Tsoporis JN,Marks A,Haddad A,Dawood F,Liu PP,Parker TGS100B expression modulates left ventricular remodeling after myocardial infarction in mice.Circulation.2005;l 11:598-606.
[16]Most P,Remppis A,Pleger ST,Loffler E,Ehlermann P,Bernotat J,Kleuss C,Heierhorst J,Ruiz P,Witt H,Karczewski P,Mao L,Rockman HA,Duncan SJ,Katus HA,Koch WJ. Transgenic overexpression of the Ca2+-binding protein S100A1 in the heart leads to increased in vivo myocardial contractile performance. J Biol Chem. 2003;278:33809-33817.
[17]Du X-J, Autelitano DJ, Dilley RJ, et al. β2-Adrenergic receptor overexpression exacerbates development of heart failure after aortic stenosis. Circulation. 2000,
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[1]Most P,Pleger ST,Volkers M,et al.Cardiac adenoviral S100A1 gene delivery rescues failing myocardium[J].J Clin Invest,2004;114(11):1550-1563.
[2]Hunt SA,Abraham WT,Chin MH,et al,ACC/AHA 2005 Guideline Update for the Diagnosis and Management of Chronic Heart Failure in the Adult.A Report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines(Writing Committee to Update the 2001 Guidelines for the Evaluation and Management of Heart Failure)[J].Circulation,2005,112(12):e154-e235.
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[13]高连如.干细胞移植治疗缺血性心脏病的进展及其作用机制[J].中国组织工程研究与临床康复,2007,11(46),9346-9352.
[14]Most P,Pleger ST,Volkers M,et al.Cardiac adenoviral S100A1 gene delivery rescues failing myocardium[J].J Clin Invest.2004;114(11):1550-1563.
[15]Pleger ST,Most P,Boucher M,et al.Stable Myocardial-specific AAV6-S100A1 gene therapy results in chronic functional heart failure rescue[J].Circulation.2007;115(19):2506-2515
[16]Kraitchman DL,Heldman AW,Atalar E,et al.In Vivo Magnetic Resonance Imaging of Mesenchymal Stem Cells in Myocardial Infarction[J].Circulation,2003,107(18):2290-2293.
[17]Arbab AS,Bashaw LA,Miller BR,et al.Characterization of Biophysical and Metabolic Properties of Cells Labeled with Superparamagnetic Iron Oxide Nanoparticles and Transfection Agent for Cellular MR Imaging[J].Radiology,2003,229(3):838-846.
[1]Pleger ST,Most P,Boucher M,Soltys S,Chuprun JK,Pleger W,Gao E,Dasgupta A,Rengo G,Remppis A,Katus HA,Eckhart AD,Rabinowitz JE,Koch WJ.Stable Myocardial-specific AAV6-S100A1 gene therapy results in chronic functional heart failure rescue.Circulation.2007;115:2506-2515.
[2]Most P,Pleger ST,Volkers M,Heidt B,Boerries M,Weichenhan D,Loffier E,Janssen PM,Eckhart AD,Martini J,Williams ML,Katus HA,Remppis A,Koch WJ.Cardiac adenoviral S100A1 gene delivery rescues failing myocardium.J Clin Invest.2004;114:1550-1563.
[3]Donato R.Intracellular and extracellular roles of S100 proteins.Microsco Res Tech.2003;60:540-551.
[4]Kiewitz R,Lyons GE,Schafer BW,Heizmann CW.Transcriptional regulation of S100A1 and expression during mouse heart development.Biochimica et Biophysica Acta.2000; 1498:207-219.
[5]Du XJ,Cole TJ,Tenis N,Gao XM, Kontgen F,Kemp BE,Heierhorst J. Impaired cardiac contractility response to hemodynamic stress in S100A1-deficient mice.Mol Cell Biol.2002; 22;2821-2829.
[6]Volkers M.Loughrey CM,Macquaide N,Reppis A,Degeorge B,Wegner F,Friedrich O,Fink RH,Koch WJ,Smith GL,Most P.S100A1 decreases calcium spark frequency and alters their spatial characteristics in permeabilized adult ventricular cardiomyocytes.Cell Calcium.2007; 41:135-143.
[7]Remppis A,Greten T,Schafer BW,Hunziker P,Erne P,Katus HA,Heizmann CW. Altered expression of the Ca(2+)-binding protein S100A1 in human cardiomyopathy. Biochim BiophysActa.1996;1313:253-257.
[8]Most P,Seifert H,Gao E,Funakoshi H,Volkers M,Heierhorst J,Remppis A,Pleger ST,DeGeorge BR Jr,Eckhart AD,Feldman AM.Koch WJ.Cardiac S100A1 protein levels determine contractile performance and propensity towards heart failure after myocardial infarction. Circulation. 2006; 114:1258 -1268.
[9]Hasenfuss,G.Alterations of calcium-regulatory proteins in heart failure. Cardiovasc Res. 1998; 37:279-289.
[10]Kiewitz R,Acklin C, Schafer BW,Maco B,Uhrik B,Wuytack F,Erae P, Heizmann CW.Ca2+-dependent interaction of S100A1 with the sarcoplasmic reticulum Ca2+-ATPase2a and phospholamban in the human heart. Biochem Biophys Res Commun. 2003;306:550-557.
[11]Lokuta AJ,Maertz NA,Meethal SV,Potter KT,Kamp TJ,Valdivia HH,Haworth RA. Increased nitration of sarcoplasmic reticulum Ca2 + ATPase in human heart failure.Circulation.2005; 111:988 -995.
[12]Milting H,Lukas N,Klauke B,Korfer R,Perrot A ,Osterziel KJ, Vogt J, Peters S, Thieleczek R,Varsanyi M.Composite polymorphisms in the ryanodine receptor 2 gene associated with arrhythmogenic right ventricular cardiomyopathy.Cardiovasc Res. 2006;71:496-505.
[13]Pleger ST,Remppis A,Heidt B,Volkers M,Chaprun JK, Kuhn M,Zhou RH,Gao E, Szabo G, Weichenhan D,Muller OJ,Eckhart AD,Katus HA,Koch WJ,Most P.S100A1 gene therapy preserves in vivo cardiac function after myocardial infarction.Mol Ther.2005;12:1120-1129.
[14]Most P,Boerries M, Eicher C,Schweda C,Ehlermann P,Pleger ST,Loffler E,Koch WJ,Katus HA,Schoenenberger CA,Remppis A. Extracellular S100A1 protein inhibits apoptosis in ventricular cardiomyocytes via activation of the extracellular-regulated kinase (ERK1/2) pathway. J Biol Chem.2003;278:48404-48412.
[15]Tsoporis JN,Marks A,Haddad A,Dawood F,Liu PP,Parker TGS100B expression modulates left ventricular remodeling after myocardial infarction in mice.Circulation.2005;l 11:598-606.
[16]Most P,Remppis A,Pleger ST,Loffler E,Ehlermann P,Bernotat J,Kleuss C,Heierhorst J,Ruiz P,Witt H,Karczewski P,Mao L,Rockman HA,Duncan SJ,Katus HA,Koch WJ. Transgenic overexpression of the Ca2+-binding protein S100A1 in the heart leads to increased in vivo myocardial contractile performance. J Biol Chem. 2003;278:33809-33817.
[17]Du X-J, Autelitano DJ, Dilley RJ, et al. β2-Adrenergic receptor overexpression exacerbates development of heart failure after aortic stenosis. Circulation. 2000,