IGF-1, bFGF EXPRESSION AND VASCULAR REGENERATION IN ACUTE INFARCTED CANINE MYOCARDIUM AFTER AUTOLOGUS SKELETAL MUSCLE SATELLITE CELL IMPLANTATION

Objective To study the cell growth factor secretion and vascular regeneration in acute in-farcted myocardium after autologous skeletal muscle satellite cell implantation. Methods Autologous skeletal muscle satellite cells from adult mongrel canine were implanted into the acute myocardial infarct site via the ligated left anterior descending (LAD) artery. Specimens were harvested at 2, 4 , 8 weeks after implantation for the expression of insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor ( bFGF) and the vascular density. Results The expression of IGF-1, bFGF and the vascular density in skeletal muscle satellite cell implant group were higher than that in the control group. Conclusion The skeletal muscle satellite cells, after being implanted into the acute myocardial infarction, not only showed myocardial regeneration, but also showed the ability to secrete the cell factors, hence representing a positive effect on the regeneration of the infarcted myocardium.     

Influence of skeletal muscle satellite cells implanted into infarcted myocardium on remnant myocyte volumes

Objective To study the effects of skeletal muscle satellite cells implanted into infarcted myocardium on the volume of remnant myocytes.Methods Thirty-six adult mongrel canines were divided randomly into implantation group and control group. In the implantation group, skeletal muscle satellite cells taken from the gluteus maximus muscles of the dogs were cultured, proliferated and labeled with 4’, 6-diamidino-2-phenylindone (DAPI) in vitro. In both groups, a model of acute myocardial infarction was established in every dog. In the implantation group, each dog was injected with M199 solution containing autologous skeletal muscle satellite cells. The dogs in the control group received M199 solution without skeletal muscle satellite cells. The dogs of both groups were killed 2, 4 and 8 weeks after implantation (six dogs in a separate group each time). Both infarcted myocardium and normal myocytes distal from the infracted regions isolated were observed under optical and fluorescent microscope. Their volumes were determined using a confocal microscopy image analysis system and analyzed using SAS. A P<0.05 was considered significant.Results A portion of the implanted cells differentiated into muscle fiber with striations and were connected with intercalated discs. Cross-sectional area and cell volume were increased in normal myocardium. Hypertrophy of remnant myocytes in the infarcted site after skeletal muscle cell implantation was much more evident than in the control group. Cross-sectional area, cell area and cell volume differed significantly from those of the control group (P< 0.05). Hypertrophy of the cells occurred predominantly in terms of width and thickness, whereas cell length remained unchanged. Conclusion Skeletal muscle satellite cells implanted into infarct myocardium, could induce the hypertrophy of remnant myocyte cells in the infarcted site and could also aid in the recovery of the contractile force of the infarcted myocardium.     

THE IMPROVEMENT OF INFARCTED MYOCARDIAL CONTRACTILE FORCE AFTER AUTOLOGOUS SKELETAL MUSCLE SATELLITE CELL IMPLANTATION

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脐血间质干细胞诱导移植梗死心肌的细胞因子分泌和血管再生

目的研究脐血间质干细胞(UCB-MSCs)梗死心肌移植的胰岛素样生长因子-1(IGF-1)、血管内皮细胞生长因子(VEGF)分泌和血管再生。方法5-氮杂胞嘧啶核苷诱导分化后的UCB-MSCs经结扎的冠状动脉左前降支远端灌注移植入梗死区,2、4、8周后取标本,用免疫组化法和VanGieson染色检测细胞因子的表达和梗死区血管的再生情况。结果UCB-MSCs移植2、4、8周后,梗死区IGF-1、VEGF表达明显高于对照组(P<0.01);治疗组梗死区微血管数量明显高于对照组。结论UCB-MSCs梗死心肌移植除有心肌再生外,尚可通过细胞因子分泌对梗死心肌起到血管再生的积极作用。     

Decreased small arterial compliance with increased serum vascular endothelial growth factor-A and circulating endothelial progenitor cell in dilated cardiomyopathy

Background Evidence showed that both myocardium and blood vessels were damaged in dilated cardiomyopathy （DCM）. However, the changes in arterial compliance, serum cytokines and circulating endothelial progenitor cells （EPC）, and their correlations remain unknown.
Methods Sixty-five DCM patients and 49 healthy volunteers were studied. Both large artery compliance （C1） and small artery compliance （C2） were measured with the CVProfUor DO-2020. Quantitative enzyme-linked immunosorbent assays （ELISAs） were used to measure the levels of vascular endothelial growth factor-A （VEGF-A） and VEGF receptor 2 （VEGF-R2）. Circulating EPC was assessed by EPC colony-forming assays and flow cytometry （CD133^＋/CD34^＋cells）. Phagocytized Dil-acLDL and binded FITC-UEA-I were used to analyze endothelial lineage marker expression by immunofluorescence.
Results Although C2 was markedly lower in DCM patients than in control group （（3.8±1.8） ml/mmHg × 100 vs （5.0±2.2） ml/mmHg × 100, P〈0.0001）, there was no statistically significant difference in C1 between the two groups （P〉0.05）. Levels of VEGF-A, the numbers of colony-forming units （CFU） and the fractions of EPC were obviously higher in DCM patients than in control group （（127.6±139.5） pg/ml vs （58.8±42.9） pg/ml, P〈0.0001; （2.5±1.5）% vs （0.5±0.3）%, P〈0.05; 23.5±12.8 vs 10.8±7.4, P〈0.01, respectively） and however, there was no significant difference in VEGF-R2 between two groups （P〉0.05）. LgVEGF-A was positively correlated with the number of EPC-CFU （r=-0.435; P〈0.05） and inversely correlated with C2 （r=-0.543; P〈0.001） in DCM patients. Conclusions The reduction of C2, a sensitive marker reflecting endothelial dysfunction, was observed in DCM patients and closely related to the increase in serum VEGF-A.     

Low-power Helium-Neon laser irradiation enhances the expression of VEGF in murine myocardium

Inrecentyears ,low powerhelium neon (He Ne)lasershavebeenincreasinglywidelyappliedinthetreatmentofcardiovasculardiseases ,anditsvasodilationeffecthasbeenproven Inourpreviousexperimentonlow powerHe Nelaserirradiationatdosesof 1 0 3,2 0 6 ,and 6 0 5J/cm2etc ,weshowedthatadoseof 6 0 5J/cm2 wassufficientlyeffectivetoinducetheameliorationofrathemorrheologicalindices1　andtheenhancementoftheexpressionofendothelialnitricoxidesynthase (eNOS)inmyocardialmicrovesselendothelium 2 　Clinicall…     

Vascular endothelial growth factor gene transfer improves host endothelializat ion of xenogeneic biologic heart valve in vivo

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Diabetic retinopathy: VEGF, bFGF and retinal vascular pathology

Background Previous research indicated that the development of diabetic retinopathy ( DR) is closely related to the excessive expression of growth factors. This paper was to study the relationship of DR with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the retinal vascular pathological change.Methods Fifty-five Wistar rats, weighing 100 -200 g, were selected and randomly divided into four groups: control group (no streptozocin injection, n =10), M1 group (streptozocin induced diabetes for 1 month, n =15), M3 group (streptozocin induced diabetes for 3 months, n =15), and M5 group (streptozocin induced diabetes for 5 months, n =15). In situ hybridization and immunohistochemistry were used to investigate the expressions of bFGF and VEGF on retinal vascular, and retinal vessels were observed by transmission electron microscope.Results There was no difference in the number of pericytes between M1 and control group (P > 0. 05), but the number of pericytes decreased obvi     

自体骨骼肌卫星细胞移植对心肌梗死大鼠心功能的影响

目的 了解自体骨骼肌卫星细胞(satellite cell,SC)移植至心肌对心肌梗死(myocardial infarction,MI)大鼠心功能的影响及其可能机制.方法 45只Wistar大鼠采用随机抽签法分为假手术组、对照组及移植组,对照组及移植组大鼠经结扎冠状动脉前降支建立MI模型.将体外培养2周的大鼠自体SC以注射的方式移植到移植组大鼠梗死区周围,4周后测定各组大鼠血流动力学、心功能、血清血管内皮生长因子(vascular endothelial growth factor,VEGF)浓度及缺血心肌毛细血管密度的变化,同时观察移植细胞在梗死区的生长、增殖情况并探讨它们相互的关系.结果 SC在梗死区中可增殖分化为横纹肌纤维;与假手术组比较,尽管对照组及移植组大鼠之收缩压、舒张压、平均动脉压、左室收缩压及左室压力最大上升/下降速率均明显降低(P＜0.05,P＜0.01),左室舒张末压均明显增高(P＜0.01),但移植组左室压力最大上升速率、左室压力最大下降速率及左室舒张末压较之对照组则有明显改善(P＜0.05,P＜0.01);SC移植4周后,对照组大鼠毛细血管密度较之假手术组明显增高(P＜0.05);移植组大鼠缺血心肌中毛细血管密度及血清VEGF浓度较之假手术组、对照组亦明显升高(P＜0.05,P＜0.01).结论 SC在心肌梗死区中能增殖分化为横纹肌样细胞,并可分泌VEGF促使缺血心肌毛细血管增生,从而共同参与改善心功能.     

Effects of Tongxinluo-facilitated cellular cardiomyoplasty with autologous bone marrow-mesenchymal stem cells on postinfarct swine hearts

Background  Treatment of ischemic heart disease remains an important challenge, though there have been enormous progresses in cardiovascular therapeutics. This study was conducted to evaluate whether Tongxinluo (TXL) treatment around the transplantation of mesenchymal stem cells (MSCs) can improve survival and subsequent activities of implanted cells in swine hearts with acute myocardial infarction (AMI) and reperfusion.
Methods  Twenty-eight Chinese mini-pigs were divided into four groups including a control group (n=7); group 2, administration of low-dose TXL alone from the 3rd day prior to AMI to the 4th day post transplantation (n=7); group 3, MSCs alone (n=7) and group 4, TXL + MSCs (n=7). AMI models were made by occlusion of the left anterior descending coronary artery for 90 minutes. Autologous bone marrow-MSCs (3×10⁷ cells/animal) were then injected into the post-infarct myocardium immediately after AMI and reperfusion. The survival and differentiation of implanted cells in vivo were detected by immunofluorescent analysis. The data of cardiac function were obtained at baseline (1 week after transplantation) and endpoint (6 weeks after transplantation) by single photon emission computed tomography (SPECT) and magnetic resonance imaging (MRI). Apoptosis was detected by TUNEL assay and the oxidative stress level was investigated in the post-infarct myocardium at endpoint.
Results  At endpoint, there was less fibrosis and inflammatory cell infiltration with more surviving myocardium in group 4 than in the control group. In group 4 the survival and differentiation of implanted MSCs were significantly improved more than that seen in group 3 alone (P<0.0001); the capillary density was also significantly greater than in the control group, group 2 or 3 both in the infarcted zone (P<0.0001) and the peri-infarct zone (P<0.0001). MRI showed that parameters at baseline were not significantly different between the 4 groups. At endpoint, regional wall thickening and the left ventricular ejection fraction were increased while the left ventricular mass index, dyskinetic segments and infarcted size were decreased only in group 4 compared with control group (P<0.0001). SPECT showed that the area of perfusion defect was significantly decreased at endpoint only in group 4 compared with control group (P<0.0001). TUNEL assay indicated that TXL administration significantly decreased cell apoptosis in peri-infarct myocardium in groups 2 and 4. Furthermore, superoxide dismutase (SOD) significantly increased and malondialdehyde (MDA) decreased in groups 2 and 4 by the administration of TXL.
Conclusions  Our study demonstrates the following: (1) immediate intramyocardial injection of MSCs after AMI and reperfusion resulted in limited survival and differentiation potential of implanted cells in vivo, thus being incapable of beneficially affecting post-hearts; (2) TXL-facilitation resulted in a significant survival and differentiation potential of implanted cells in vivo via inhibition of apoptosis and oxidative stress, accompanied by significant benefits in cardiac function.

     

Alleviation of hypoxic pulmonary vascular structural remodeling by L-arginine

目的探讨L-精氨酸(L-Arg)对低氧性肺血管结构重建的干预作用及其可能机制.方法将18只Wistar大鼠配伍后随机分为对照组、低氧组和低氧+L-Arg组(共6个配伍组).以右心导管法测定肺动脉压力,并对大鼠肺组织标本进行显微结构观测和超微结构观察,同时以分光光度法测定血浆一氧化氮(NO)含量,并对肺组织以内皮素-1(ET-1) cRNA探针进行原位杂交,研究肺动脉内皮细胞ET-1 mRNA的表达.结果低氧组大鼠肺动脉平均压明显高于对照组(20.33±2.18?mm?Hg vs 15.38±1.05?mm?Hg, P<0.05).低氧后大鼠肺血管显微及超微结构发生明显改变,肺血管结构重建形成.同时低氧组大鼠血浆NO间接含量明显低于对照组 (P<0.05 ).低氧后肺动脉内皮细胞ET-1 mRNA表达明显增强.然而,低氧+L-Arg组大鼠PAMP较低氧组明显降低(16.73±1.35?mm?Hg vs 20.33±2.18?mm?Hg, P<0.05).L-Arg缓解了低氧性肺血管结构重建的形成.同时低氧+L-Arg组大鼠血浆NO间接含量明显高于低氧组(P<0.05).L-Arg使低氧大鼠肺动脉内皮细胞ET-1 mRNA表达明显受抑制.结论 L-Arg对低氧性肺血管结构重建以及低氧性肺动脉高压的形成有重要的调节作用,其机制可能与通过促进低氧大鼠体内NO生成,从而抑制肺动脉内皮细胞ET-1 mRNA表达有一定的关系.     

骨骼肌卫星细胞移植对残存心肌细胞游离Ca~(2 )的影响

目的研究骨骼肌卫星细胞移植对梗塞心肌残存心肌细胞游离Ca2 分布及Ca2 波的恢复作用。方法自犬臀部取骨骼肌 ,提取、培养、扩增骨骼肌卫星细胞 ,4’ ,6-diamidino -2 -phenylindone(DAPI)荧光标记 ,经结扎的左冠状动脉前降支远端灌注移植入自体急性心肌梗塞区域 ,2、4、8周后处死动物 ,取心肌标本行组织形态学、心肌细胞游离Ca2 分布及Ca2 波检测。　结果梗塞区观察到带荧光的细胞核及肌纤维 ,部分已分化成横纹肌并以闰盘相连。梗塞区残存心肌细胞游离Ca2 增加 ,Ca2 波峰出现明显晚于正常区域心肌细胞 ,且峰值低于正常心肌细胞。骨骼肌卫星细胞移植 8周后 ,梗塞区残存心肌细胞游离Ca2 分布及Ca2 波峰恢复正常。　结论经冠状动脉灌注移植骨骼肌卫星细胞使梗塞心肌残存心肌细胞游离Ca2 分布及Ca2 波恢复 ,利于心肌收缩力的恢复。     

Vascular endothelial growth factor gene transfection to enhance the repair of avascular necrosis of the femoral head of rabbit

Avascularnecrosisofthefemoralheadisadebilitatingdiseasethatusuallyleadstodestructionofthehipjointandincreasingmusculoskeletalmorbidityinyoungandmiddleagedpatients Totaljointreplacementiscommonlyusedtotreatdisablingsecondaryosteoarthrosis Althoughthemostrecentimprovementsintotalhipreplacementmaydecreasefailure,thepatientsareusuallyyoungandcurrenthipprostheseswon’tfunctionwellfortheremaininglifeexpectancyofthesepatients Thus,it’snecessarytodelayoreliminatetheneedforhipreplacementbytreatingthis…     

血管内皮生长因子在前列腺癌中的表达及与肿瘤微血管密度和肿瘤细胞增殖的关系

目的探讨血管内皮生长因子(VEGF)在前列腺癌中的表达及与肿瘤微血管密度(MDV)和肿瘤细胞增殖的关系。方法采用免疫组织化学S-P法测定42例前列腺癌组织和10例正常前列腺组织中VEGF、KI-67的表达和MVD计数。结果前列腺癌中VEGF的阳性表达率为77.78%,KI-67LI和MVD计数分别为12.67±4.12和37.23±9.76,VEGF表达水平与肿瘤恶性程度、KI-67标记指数和MDV计数呈正相关。结论VEGF有促进肿瘤细胞增殖及血管生成的作用,联合检测VEGF、KI-67和CD34可作为病理诊断的补充,对患者预后判断和治疗方案的选择有重要参考价值。     

骨骼肌卫星细胞移植对残存心肌细胞游离Ca2+的影响

目的研究骨骼肌卫星细胞移植对梗塞心肌残存心肌细胞游离Ca2+分布及Ca2+波的恢复作用。方法自犬臀部取骨骼肌,提取、培养、扩增骨骼肌卫星细胞,4’,6-diamidino-2-phenylindone(DAPI)荧光标记,经结扎的左冠状动脉前降支远端灌注移植入自体急性心肌梗塞区域,2、4、8周后处死动物,取心肌标本行组织形态学、心肌细胞游离Ca2+分布及Ca2+波检测。结果梗塞区观察到带荧光的细胞核及肌纤维,部分已分化成横纹肌并以闰盘相连。梗塞区残存心肌细胞游离Ca2+增加,Ca2+波峰出现明显晚于正常区域心肌细胞,且峰值低于正常心肌细胞。骨骼肌卫星细胞移植8周后,梗塞区残存心肌细胞游离Ca2+分布及Ca2+波峰恢复正常。结论经冠状动脉灌注移植骨骼肌卫星细胞使梗塞心肌残存心肌细胞游离Ca2+分布及Ca2+波恢复,利于心肌收缩力的恢复。     

Affects of different access routes on autologous satellite cell implantation stimulating myocardial regeneration

Objective To study the effect of different access routes on autologous satellite cell implantation to stimulate myocardial regeneration.Methods Satellite cells were procured from skeletal muscle (gluteus max) of adult mongrel canine, cultured, proliferated and labeled with 4’,6-diamidino-2-phenylindone (DAPI) in vitro.The cells were autologously implanted into the site of acute myocardial infarction by local injection or perfusion through the ligated distal left anterior descending coronary artery.Specimens were harvested 2, 4 and 8 weeks later for histological study. Results The labeling efficiency of satellite cells with DAPI was close to 100%.Fluorescent cells were found at the infarcted zone, papillary muscle and local injection site.Some of these cells had progressively differentiated into striated muscle fibers connected to intercalated discs.The infant cells appeared different from the mature myocardium under an electron microscope.Satellite cells implanted by perfusion through the coronary artery were arranged in order of consistency with host myocardial fibers.The satellite cells, implanted by local injection, were found growing in a disordered way. Conclusion Satellite cells, implanted by coronary artery perfusion, can progressively differentiate into striated muscle fibers, arranging in order and disseminating over the infarcted zone.This approach seems more favorable for the recovery of myocardial contractile function than that of local injection.     

家兔骨骼肌缺血预处理后血管内皮生长因子的表达

目的　探讨血管内皮生长因子 (VEGF)与缺血预处理 (IPC)之间的关系 ,为IPC的内源性保护作用机理提供实验依据。方法　取健康兔 2 4只 ,随即分成三组 :缺血—再灌注组 (I/R组 )、IPC组和对照组 ,每组 8只。采用免疫组织化学方法检测各骨骼肌细胞VEGF表达的阳性率。结果　兔骨骼肌细胞VEGF表达阳性率I/R组为 5 8.75 % ,IPC组为49.2 5 % ,对照组为 2 9.75 %。I/R组和IPC组VEGF表达明显强于对照组 ( χ2 分别为 6 8.18,31.82 ,P均 <0 .0 0 1) ,IPC组VEGF表达弱于I/R组 ,χ2 =7.2 7,P <0 .0 1。结论　VEGF参与IPC的延迟保护作用。     

Effect of arsenic trioxide on inhibition of restenosis after rabbit vascular injury and its mechanism

目的　探讨应用三氧化二砷 (As2 O3)预防血管损伤后再狭窄的效果及其作用机理。方法　观察As2 O3对培养兔血管平滑肌细胞 (VSMCs)凋亡的影响。 32只新西兰白兔随机分为 2、4周实验组和对照组 ,分别 10 %As2 O32 5mg·Kg 1·d 1或等量生理盐水腹腔注射 3天 ,应用球囊损伤左颈总动脉。处死动物取血管作形态学和免疫组化检测 ,并检查肝脏、肾脏组织学变化。结果　细胞形态学和DNA电泳梯形带证实 ,As2 O3诱导培养VSMCs凋亡 ,与药物浓度和作用时间呈依赖性。与对照组相比 ,2 wk实验组血管内膜增殖面积显著减少 (P <0 0 5 ) ,4 wk组内膜面积无明显差异 ;但 2 ,4 wk组的管腔面积均有明显增大 (P均 <0 0 5 )。与对照组相比 ,实验组 2 ,4 wk免疫组化显示bcl 2表达下调 (P均 <0 0 5 ) ,Bax表达上调 (P分别 <0 0 1,0 0 5 ) ,均与相对应的血管内膜增生受抑制 ,血管腔面积扩大相吻合结论　As2 O3诱导VSMCs凋亡和有效预防实验性血管损伤后再狭窄 ,均与下调bcl 2和上调bax表达有关     

Extracellular domain of kinase domain region mediated by adeno-associated virus inhibits growth and angiogenesis of bladder cancer in Balb-c mice

目的：从原代培养的人脐静脉血管内皮细胞（HUVEC）提取细胞总RNA,采用逆转录PCR(RT-PCR)方法得到VEGF受体KDR全长胞外区cDNA片段,检验其体内抗肿瘤血管生成的作用。方法：将获得的受体基因克隆到AAV基因治疗载体pSNAV中,得到重组质粒pSNAV/KDR。重组质粒转染BHK细胞,加入辅助病毒后,得到表达目的蛋白的重组AAV。重组AAV表达的KDR具有与VEGF结合的活性,重组AAV感染膀胱癌EJ细胞,皮下注射Balb-c裸鼠。Ⅷ因子免疫组化染色进行微血管密度测定。结果：重组AAV感染有膀胱癌EJ细胞形成的肿瘤血管化程度明显低于对照组。结论：重组AAV介导的KDR胞外区基因可有效的抑制裸鼠人膀胱癌组织的新生血管生成。