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RONG Qi-fei HUANG Jun SU En-ben LI Jun LI Jian-yong ZHANG Li-li CHEN Lei-lei WANG Xiao-bin CAO Ke-jiang 《中华医学杂志(英文版)》2008,121(3):248-256
Background Hepatitis B virus (HBV) replication has been reported to be involved in many extrahepatic viral disorders; however, the mechanism by which HBV is transinfected into extrahepatic tissues such as myocardium and causes HBV associated myocarditis remains largely unknown.
Methods In this study, endothelial progenitor cells (EPCs) were infected by HBV and then transfused into ischemic model of mice. HBV surface and core antigen as well as mutation of HBV particles were detected by immunohistochemistry, fluorescent activated cell sorter and transmission electron microscopy in vitro and in vivo.
Results Human cord blood EPCs, but not human umbilical vein endothelial cells (HUVECs) could be effectively infected by taking up HBV in vitro. HBV envelope surface and core antigen expressions were first detectable in EPCs at day 3 after virus challenge, sustained for up to 11 days, and decreased thereafter. Similarly, the virus particles were the most abundant in EPCs in the first week observed by a transmission electron microscope, and declined in 3 weeks after HBV infection. HBV DNA but not HBV cccDNA in EPCs were detectable even 3 weeks after virus challenge, as shown by PCR analysis. Furthermore, intravenous transplantation of HBV-treatod EPCs into myocardial infarction Sprague & Dawley rats model resulted in incorporation of both EPCs and HBV into injured endothelial tissues of capillaries in the ischemic border zone.
Conclusions These results strongly support that EPCs serve as virus carrier mediating HBV trans-infection into the injured myocardial tissues. The findings might suggest a novel mechanism for HBV-associated myocarditis. 相似文献
Methods In this study, endothelial progenitor cells (EPCs) were infected by HBV and then transfused into ischemic model of mice. HBV surface and core antigen as well as mutation of HBV particles were detected by immunohistochemistry, fluorescent activated cell sorter and transmission electron microscopy in vitro and in vivo.
Results Human cord blood EPCs, but not human umbilical vein endothelial cells (HUVECs) could be effectively infected by taking up HBV in vitro. HBV envelope surface and core antigen expressions were first detectable in EPCs at day 3 after virus challenge, sustained for up to 11 days, and decreased thereafter. Similarly, the virus particles were the most abundant in EPCs in the first week observed by a transmission electron microscope, and declined in 3 weeks after HBV infection. HBV DNA but not HBV cccDNA in EPCs were detectable even 3 weeks after virus challenge, as shown by PCR analysis. Furthermore, intravenous transplantation of HBV-treatod EPCs into myocardial infarction Sprague & Dawley rats model resulted in incorporation of both EPCs and HBV into injured endothelial tissues of capillaries in the ischemic border zone.
Conclusions These results strongly support that EPCs serve as virus carrier mediating HBV trans-infection into the injured myocardial tissues. The findings might suggest a novel mechanism for HBV-associated myocarditis. 相似文献
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目的 探讨犬脐血血管内皮祖细胞(EPCs) 移植对心肌梗死血管形成的影响.方法 取妊娠犬脐血,体外分离、培养、扩增EPCs,免疫组化鉴定.建立成年杂种犬梗死模型,经BrdU标记的EPCs灌注移植入梗死区域,1、4、8周后处死动物,取心肌标本HE染色确认梗死模型、免疫组化染色BrdU观察EPCs参与梗死心肌血管形成、vW因子染色观察梗死心肌血管并计数以观察EPCs移植组与对照组血管形成差异.结果 免疫组化检测结果表明培养的细胞为EPCs;HE染色示心肌梗死区有大量瘢痕组织、成纤维细胞及小血管形成;免疫组化检测显示梗死心肌区域标本内小血管上存在BrdU阳性细胞;EPCs移植组与对照组心肌梗死后第1、4、8周的心肌缺血区和梗死区的血管计数均无显著差异.结论 犬脐血EPCs梗死心肌移植可参与血管形成,但不能促进心肌的血管再生. 相似文献
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Transplanted human umbilical cord blood mononuclear cells improve left ventricular function through angiogenesis in myocardial infarction 总被引:9,自引:1,他引:8
Background Human umbilical cord blood contains an abundance of immature stem/progenitor cells, which may participate in the repair of hearts that have been damaged by myocardial infarction (MI). This study aimed to evaluate the effects of human umbilical cord blood mononuclear cells (hUCBC) transplantation on cardiac function and left ventricular remodeling in rat model of MI. Methods Forty-five male Wistar rats were randomized into three groups: MI or control group (n=15), MI plus cell transplantation (n=15), and sham group (n=15). Acute myocardial infarction (AMI) was established by ligating the left anterior descending artery, thereafter, hUCBC were implanted into the marginal area of infarcted myocardium. In MI/control group, DMEM was injected instead of hUCBC following the same protocol. Left ventricular function assessment was carried out by echocardiography and invasive hemodynamic measurements one month post MI. All rats were sacrificed for histological and immunochemical examinations. Results The transplanted hUCBC survived and engaged in the process of myocardial repair in the host heart. Echocardiography demonstrated that left ventricular function improved significantly in the rats that underwent cell transplantation. Hemodynamic studies found a significantly decreased left ventricular end-diastolic pressure (LVEDP) [(21.08±8.10) mmHg vs (30.82±9.59) mmHg, P<0.05], increase in +dp/dt(max) [(4.29±1.27) mmHg/ms vs (3.24±0.75) mmHg/ms, P<0.05), and increase in -dp/dt(max) [(3.71±0.79) mmHg/ms vs (3.00±0.49) mmHg/ms, P<0.05] among MI group with hUCBC transplantation when compared with MI/control group. Masson’s trichrome staining revealed that the collagen density in the left ventricle was significantly lower in rats of transplantation group than that in the MI control groups [(6.33±2.69)% vs (11.10±3.75)%, P< 0.01]. Based on immunostaining of α-actin, the numbers of microvessels were significantly (P<0.01) increased at the boundary of infarction site. Similarly higher mRNA expression of vascular endothelial growth factor (VEGF) 164 and VEGF188 were found at 7- and 28-day post cell transplantation in MI group with hUCBC transplantation when compared with MI/ control group.Conclusions Transplanted hUCBC can survive in host myocardium without immunorejection, significantly improve left ventricular remodeling after AMI and promote a higher level of angiogenesis in the infarct zones. All these factors beneficially affect cardiac repair in the setting of MI. Therefore human umbilical cord blood may be potential source for cell-based therapy for AMI. 相似文献
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Optimal time for human umbilical cord blood cell transplantation in rats with myocardial infarction 总被引:1,自引:0,他引:1
XING Yun-li SHEN Lu-hua LI Hong-wei ZHANG Yu-chen ZHAO Lin ZHAO Shu-mei XU Qing 《中华医学杂志(英文版)》2009,122(23):2833-2839
Background Cell therapy for cardiac regeneration is still under investigation. To date there have been a limited number of studies describing the optimal time for cell injection. The present study aimed to examine the optimal time for human umbilical cord blood cells (HUCBCs) transplantation after myocardial infarction (MI). Methods The animals underwent MI by ligation of the left anterior descending coronary artery and received an intravenous injection of equal volumes of HUCBCs or phosphate buffered saline at days 1, 5, 10 and 30 after MI. HUCBCs were detected by immunostaining against human human leucocyte antigen (HLA). Cardiac function, histological analysis and measurement of vascular endothelial growth factor (VEGF) were performed 4 weeks after cell transplantation. Results HUCBCs transplantation could improve cardiac function in rats that received transplantation at 5 and 10 days after MI. The best benefit was achieved in rats that received cells at 10-day after MI. Survival of engrafted HUCBCs, angiogenesis and VEGF expression were more obvious in the 10-day transplantation group than in the other transplantation groups. No evidence of cardiomyocyte regeneration was detected in any transplanted rats. Conclusions HUCBCs transplantation could improve cardiac function in rats that received HUCBCs at days 5 and 10 after MI with the optimal time for transplantation being 10 days post MI. Angiogenesis, but not cardiomyocyte regeneration, played a key role in the cardiac function improvement. 相似文献
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[目的]探讨人脐血单个核细胞移植对急性心肌梗死后血管再生的作用.[方法]雄性Wistar大鼠60只随机分为对照组(心肌梗死)和移植组(心肌梗死+细胞移植)各30只.结扎冠状动脉左前降支制作大鼠急性心肌梗死模型,以羟乙基淀粉沉淀加密度梯度离心的方法制备人脐血单个核细胞,并以5-溴脱氧尿核苷(BrdU)标记细胞.移植组大鼠模型制作成功后即在梗死区周边注射经分离并标记的人脐血单个核细胞混悬液(2×10^5/μL),在对照组相同位置注入等量达尔伯克必需培养基(DMEM).移植4周后用左心导管检测血流动力学改变,并取心脏组织行抗第Ⅷ因子(vWF)和BrdU免疫组化染色,观察毛细血管密度、移植细胞成活情况.并分别在移植后第4天、第7天、第14天、第28天取梗死周边区组织作血管内皮生长因子(VEGF)RT-PCR的半定量研究.[结果]移植组移植的单个核细胞可在梗死心肌内存活.移植组比对照组心功能明显改善,左心室舒张末压(LVEDP)明显降低(21.08±8.10)mmHg vs(30.82±9.59)mmHg,P<0.05;左心室内压力最大上升速率(+dp/dtmax)明显增快(4.29±1.27)mmHg/ms vs(3.24±0.75)mmHg/ms,P<0.05;最大下降速率(-dp/dtmax)亦显著提高(3.71±0.79)mmHg/ms vs(3.00±0.49)mmHg/ms,P<0.05.移植组梗死区周边毛细血管数显著增加(5.7±0.3)/HP vs(2.3±0.4)/HP,P<0.01.VEGF mRNA表达在移植组于第7天明显增加,并持续至第28天,对照组于第4天及第7天可见少量表达,第14天与第28天明显减弱,两组差异有统计学意义(P<0.01).[结论]人脐血干细胞在未使用免疫抑制剂的条件下可成功移植到大鼠急性心肌梗死区,改善急性心肌梗死大鼠心功能,促进新生血管形成. 相似文献
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目的 观察犬脐血内皮祖细胞(endothelial progenitor cells,EPCs)急性梗死心肌移植对心肌胶原网络重构的影响.方法 取足月妊娠犬脐血,体外分离培养、扩增EPCs,免疫组化鉴定,成年杂种犬于冠状动脉前降支第一对角支分出后结扎建立梗死模型,经BrdU标记的EPCs灌注移植入结扎的冠状动脉远端梗死区域,1、4、8周后处死动物,取急性心肌梗死区域标本检测.结果 4~6 d可见细胞贴壁生长,7~14 d可见贴壁细胞逐渐融合并呈克隆型生长,呈"铺路石"样外观,细胞标记CD31 ,vW因子 ,FLK-1 ,CD34 ;HE染色结果显示,急性心肌梗死区有大量瘢痕组织、成纤维细胞及小血管形成;BrdU染色结果显示,小血管上见BrdU阳性细胞;第1、4、8周脐血EPCs移植组和对照组急性心肌梗死区血管计数差异均无统计学意义(5.05±0.89、15.56±0.99、12.06±1.00 vs 4.89±0.84、15.67±0.98、12.50±0.99,P均>0.05);VG染色结果显示,脐血EPCs移植组和对照组急性心肌梗死后第1、4周胶原纤维含量较少,第8周胶原纤维排列趋于规则,心肌纤维较少,脐血EPCs移植组急性心肌梗死后第1周IOD值与对照组之间差异无统计学意义(586.8±120.9 vs 599.6±156.2,P>0.05),脐血EPCs移植组第4、8周IOD值均明显低于对照组(588.2±249.0、971.5±288.5 vs 1 206.2±299.4、1 823.8±329.7,P均<0.05).结论 犬脐血EPCs急性梗死心肌移植可参与血管形成,尽管不能促进心肌的血管再生,但对心肌胶原网络重构有改善作用. 相似文献
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目的研究脐血间充质干细胞(MSCs)移植对心肌梗死区域旁残存心肌组织的影响。方法自孕犬脐血中分离、培养MSCs,经Laz基因转染和5-氮胞苷体外肌源性诱导后移植入犬急性心肌梗死模型(移植组,n=18)。分别于移植后1、4、8周处死动物,取心肌组织标本行TUNEL染色观察残存细胞凋亡情况并计算凋亡指数;应用激光共聚焦扫描显微镜图像分析系统检测残存心肌细胞体积;Nagar-Olsen染色观察心肌胶原网络的变化。以等量生理盐水注射作为移植对照组(n=18)。结果与对照组比较,移植组缺血区部各时间点切片位观察到凋亡细胞数目显著减少,凋亡指数明显降低;残存心肌细胞肥大明显;移植后第8周,心肌组织内胶原纤维排列整齐、规律。结论脐血MSCs移植可通过减少心肌细胞凋亡、促使残存心肌细胞肥大、调节心肌胶原网络而改善心功能。 相似文献
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目的:观察体内心肌缺血微环境诱导下人脐血间充质干细胞(umbilical cord blood mesenchymal stem cells,UCBMSCs)向心肌样细胞的定向分化及对急性心肌梗死(acute myocardial infarction,AMI)大鼠心功能、新生血管的影响。方法:收集健康产妇脐血细胞,采用密度梯度离心结合差速贴壁的方法分离MSCs;健康成年SD大鼠30只,制备成AMI大鼠模型后分为2组,移植组在心肌梗死周边区注射移植GFP标记的UCBMSCs;对照组在心肌梗死周边区注射等量生理盐水。术后4周行超声心动图检测及血流动力学检查,并取心脏组织行冰冻切片,免疫荧光染色检测UCBMSCs心肌特异性蛋白cTnT和Connexin43的表达;并进行抗Ⅷ因子抗体免疫组化染色,观察心肌毛细血管密度(myocardial capillary density,MCD)的变化。结果:与对照组相比,术后4周移植的UCBMSCs表达心肌特异性蛋白cT-nT和Connexin43,对照组无心肌特异蛋白表达;移植组LVEDD、LVESD明显减小,而LVEF、LVFS明显增加,血流动力学指标明显改善;免疫组化染色结果显示,移植组梗死周边区MCD较对照组明显增加,移植组为(4.16±0.2)个/HP(×400),对照组为(2.29±0.3)个/HP,两组比较差异有统计学意义(P<0.01)。结论:UCBMSCs移植可在大鼠AMI部位存活,并向心肌样细胞分化;UCBMSCs移植后明显改善AMI大鼠心功能及心室重构,促进毛细血管新生。 相似文献
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Objective This review discussed the available data on treatment outcomes of cord blood transplantation (CBT) for acute leukemia.Data sources The data cited in this review were obtained from articles li... 相似文献
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目的观察人脐带血干细胞(HUCBSC)移植对大鼠急性心肌梗死后左室重构的影响。方法采用Ficoll-Hypague法分离培养HUCBSC;健康雄性Wistar大鼠40只,随机均分为对照组和实验组,分别将脐带血和HUCBSC经心外膜下植入对照组和实验组梗死心肌周围。结果移植术后4周,实验组左心室非梗死区心肌骨桥蛋白基因(OPN mRNA)、Ⅰ型胶原和AngⅡ含量均显著低于对照组(P〈0.05)。Pearson直线相关分析表明,实验组和对照组心肌AngⅡ、Ⅰ型胶原与OPN mRNA三者之间两两均呈正相关(P〈0.05)。结论人脐带血干细胞移植可能通过抑制大鼠心肌Ⅰ型胶原合成,下调心肌AngⅡ和OPN基因表达,减轻左室重构。 相似文献
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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 总被引:5,自引:0,他引:5
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。 相似文献
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丹参多酚酸盐对猪急性心肌梗死后心功能及侧枝循环形成有促进作用 总被引:5,自引:0,他引:5
目的:观察丹参多酚酸盐(salvianolate)对猪急性心肌梗死的治疗作用.方法:苏中幼猪21只,随机分为高剂量、低剂量组和对照组,3组均经开胸结扎冠状动脉左前降支制作心肌梗死模型,造模成功后,两组治疗组(高剂量组:250 ml 5%GS 400 mg丹参多酚酸盐;低剂量组:250 ml 5%GS 200 mg丹参多酚酸盐)结扎后当天开始经静脉途径静滴,对照组经静脉途径应用250 ml 5%GS静滴,均为1次/天,疗程7天.结扎后4周行冠状动脉造影、门控心肌显像评价侧支循环和心功能,然后处死动物,取心脏标本,行免疫组化检查.结果:①冠状动脉造影显示:治疗组梗死区侧枝循环明显优于对照组.尤以高剂量组更为显著;②门控心肌显像:两治疗组的左室射血分数(LVEF)都高于对照组:③免疫组化结果显示:两治疗组的毛细血管密度明显大于对照组,但高剂量组与低剂量组之间无显著差异.两治疗组的功能血管密度也明显大于对照组.且高剂量组明显优于低剂量组(P<0.05).结论:静脉应用丹参多酚酸盐能够促进梗死心脏心功能改善和促进血管再生.能促进侧支血管的生成. 相似文献
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脐血来源树突状细胞的培养鉴定 总被引:2,自引:0,他引:2
目的:探索体外诱导出的成熟脐血来源树突状细胞(CBDCs)功能性抗原表达的变化规律。方法:用光镜观察培养12dCBDCs的形态。用流式细胞仪分析培养不同时间CBDCs功能性抗原表达的变化规律。结果:体外诱导出典型形态的CBDCs,HLA-ABC、HLA-DR、CD1a在培养0~8d表达率渐升高,在第12d达高峰,第16d表达有所下降;CD83、CD80在第12d的表达略高于其他时间段;CD54在0d、4d、8d的表达率相近,而第12d的表达率明显升高,第16d明显降低。结论:应用本的方法,在体外培养第12d,能诱导出典型形态的CBDCs,多种功能性抗原的表达达到高峰。 相似文献
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目的 :研究脐血造血干细胞移植治疗小儿慢性粒细胞白血病的疗效及特点。方法 :给一例 12岁男性慢性粒细胞白血病慢性期的患儿移植HLA配型基本相合的同胞脐血。预处理选用BU CY方案 (马利兰 4mg·kg- 1 × 4d ,环磷酰胺 6 0mg·kg- 1 × 2d)。预防移植物抗宿主病 (GVHD)采用环孢霉素A(CsA)。移植有核细胞数为 2 0× 10 7·kg- 1 。结果 :移植后 2 2d(+2 2d)微卫星位点分析显示移植物植入成功 ,+2 5d骨髓显示造血开始重建 ,+42d中性粒细胞 >0 .5× 10 9·L- 1 ,+91dBCR ABL融合基因表达转阴 ,且受者血型由O型转为供者血型B型。随访 15 0d ,未发生急、慢性GVHD。结论 :脐血造血干细胞移植对小儿慢性粒细胞白血病有根治性治疗作用 ,它具有GVHD发生率低、程度较轻、重建造血慢等特点。 相似文献
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呈急性心肌梗死样表现的暴发性心肌炎11例 总被引:1,自引:0,他引:1
目的分析呈急性心肌梗死样表现的暴发性心肌炎临床特点及治疗方法.方法研究对象符合①45岁以下;②无高血压、高脂血症、糖尿病、吸烟等常见的动脉粥样硬化危险因素;③既往无心绞痛表现;④心肌酶学增高(较正常参考值高2倍以上);⑤ECG除极相及(或)复极相出现较典型的AMI样改变,即出现异常Q波、ST段抬高或压低;⑥1个月内恢复正常或死亡者.从临床表现、辅助检查及治疗等方面进行分析.结果共11例入选,男性7例,女性4例,平均年龄26岁.8例有前驱感染症状,9例发生心功能不全或室速或严重的房室传导阻滞,10例患者心电图多导联出现异常.根据心超表现分为3型,Ⅰ型3例为心脏结构、心脏收缩功能及舒张功能均无明显异常;Ⅱ型4例为心室腔不增大,但室壁(室间隔或正后壁)增厚,心脏收缩功能及舒张功能均减退,但以舒张功能不全为主;Ⅲ型4例为左心室腔明显扩大,心脏收缩功能明显减退.糖皮质激素对2型效果好.存活7例,经1~2年随访逐渐恢复.结论呈急性心肌梗死样表现的暴发性心肌炎病情重、死亡率高,多有前驱感染症状,常出现多导联心电图异常,依据心超表现的分型对治疗与预后有重要意义. 相似文献