Improvement of cardiac function after transplantation of autologous bone marrow mesenchymal stem cells in patients with acute myocardial infarction

Background The infarct size determines the long-term prognosis of patients with acute myocardial infarction (AMI). There is a growing interest in repairing scar area by transplanting bone marrow stem cells. However, effectiveness of intracoronary injection of bone marrow mesenchymal stem cells (BMSCs) in patients with AMI still remains unclear.Methods Sixty-nine patients with AMI after percutaneous coronary intervention (PCI) were randomly divided into intracoronary injection of BMSCs (n=34) and saline (control group, n=35) groups. Serial single positron emission computer tomography (SPECT), cardiac echo and cardiac electromechanical mapping were done at the designed time intervals until six months after transplantation of BMSCs or injection of saline. Results The proportion with functional defect decreased significantly in the BMSCs patients after three months ［(13±5)%］ compared with that pre-transplantation ［(32±11)%］ and the control group ［(28±10)%］ at three month follow-up (P<0.05, respectively). Wall movement velocity over the infracted region increased significantly in the BMSCs group ［(4.2±2.5) cm/s vs (2.2±1.3) cm/s, P<0.05］, but not in the control group ［(2.2±1.5) cm/s vs (2.7±1.7) cm/s, P>0.05］. Left ventricular ejection fraction (LVEF) three months after transplantation in BMSCs group increased significantly compared with that pre-implantation and with that of the control group at three months post-injection ［(67±11)% vs (49±9)% and (53±8)%, P<0.05 respectively］. SPECT scan results showed that perfusion defect was improved significantly in BMSCs group at three-month follow-up compared with that in the control group ［(134±66)cm2 vs (185±87)cm2, P<0.01］. At the same time, left ventricular end-diastolic volume ［(136±31) ml vs (162±27) ml, P<0.05］ and end-systolic volume ［(63±20) ml vs (88±19) ml, P<0.05］ decreased synchronously. The ratio of end-systolic pressure to end-systolic volume ［Psyst/ESV, (2.84±1.30) mmHg/ml vs (1.72±1.23) mmHg/ml, P<0.05］ increased significantly. Cardiac electromechnical mapping demonstrated significant improvement at three months after implantation of BMSCs compared with that pre-injection in both cardiac mechanical capability as left line local shorting ［LLS, (11.29±1.64)% vs (7.32±1.86)%, P<0.05］ and electrical property as left ventricular endocardial unipolar voltage ［UV, (10.38±1.12) mV vs (7.61±1.09) mV, P<0.01］; perfusion defect decreased from (36.2±6.2) % to (20.3±5.31)% (P<0.01). Twenty-four-hour electrocardiographic monitoring demonstrated no arrhythmias occurred at three-months follow-up.Conclusions The transplantation of BMSCs might improve the cardiac function and it is safe and feasible with no deaths or malignant arrhythmias.     

Influence of Transplantation of Allogenic Bone Marrow Mononuclear Cells on the Left Ventricular Remodeling of Rat after Acute Myocardial Infarction

Ventricular remodeling after acute myocardial in-farction (AMI) not only significantly increases the complications and deaths during the acute period but also results in the progressive impairment of ventricular func-tion progress, finally leading to congestive heart failure. Therefore, restricting or reducing the ventricular remod-eling after AMI has become one of important topic in cardiovascular studies. In recent years, the study on stem cells transplantation has been making steady progr…     

间充质干细胞移植早期对心肌细胞凋亡及左心室功能的影响

目的 研究骨髓间充质干细胞(mesenchymal stem cells,MSC) 缺血心肌内移植早期对心肌细胞凋亡、血流动力学及左心室功能的影响.方法 采用密度梯度离心+ 贴壁培养方法分离培养Wistar 大鼠MSC,结扎左前降支建立急性心肌梗死(AMI)模型,结扎后30min 随机分为MSC 组( 于梗死边缘分4 点心肌内注射1×106 /0.1ml 的MSC,10 只),AMI 组( 同法心肌内注射PBS 0.1ml,10 只),假手术组(SHAM 组,5 只).监测细胞移植后72h 血流动力学、超声心动图及TUNEL 心肌细胞凋亡的变化.结果 与SHAM 组相比,AMI 大鼠血流动力学恶化,表现为平均动脉压、左心室收缩压明显下降,左心室舒张末压显著升高,左心室射血分数及缩短分数均显著降低,梗死及缺血区心肌细胞凋亡比例分别为41.6% 和18.9% ;MSC移植减少梗死及缺血区心肌细胞凋亡32% 和51%(P 均<0.01),但对大鼠的平均动脉压、左心室收缩压、左心室舒张末压及左心室功能无明显改善作用(P 均>0.05).结论 MSC 移植早期能减少心肌细胞凋亡但不改善急性心肌梗死后大鼠的心功能.     

Assessment of left ventricular segmental function after autologous bone marrow stem cells transplantation in patients with acute myocardial infarction by tissue tracking and strain imaging

Background Emerging evidence suggests that stem cells can be used to improve cardiac function in patients after acute myocardial infarction. In this randomized trial, we aimed to use Doppler tissue tracking and strain imaging to assess left ventricular segmental function after intracoronary transfer of autologous bone-marrow stem cells (BMCs) for 6 months’ follow up. Methods Twenty patients with acute myocardial infarction and anterior descending coronary artery occlusion proven by angiography were double-blindedly randomized into intracoronary injection of bone-marrow cell (treated, n=9) or diluted serum (control, n=11) groups. GE vivid 7 and Q-analyze software were used to perform echocardiogram in both groups 1 week, 3 months and 6 months after treatment. Three apical views of tissue Doppler imaging were acquired to measure peak systolic displacement (Ds) and peak systolic strain (εpeak) from 12 segments of LV walls. Left ventricular ejection fraction (LVEF), end-diastolic volume (EDV) and end-systolic volume (ESV) were obtained by Simposon’s biplane method. Results (1) 3 months later, Ds and εpeak over the infract-related region clearly increased in the BMCs group ［Ds: (4.49±2.71) mm vs (7.56±2.95) mm, P&lt;0.01; εpeak: (-13.40±6.00)% vs (-17.06±6.05)%, P&lt;0.01］, but not in the control group ［Ds: (4.74±2.67) mm vs (5.01±3.23) mm, P&gt;0.05; εpeak : (-13.84±6.05)% vs (-15.04±6.75)%, P&gt;0.05］. At the same time, Ds over the normal region also increased, but the Ds enhancement was markedly higher in the BMCs group than that in the control group ［(3.21±3.17) mm vs (0.76±1.94) mm, P&lt;0.01］. Parameters remained steady from the 3rd to 6th month in either group (P&gt;0.05). (2) LVEF in treated and control groups were almost the same at baseline (1st week after PCI) ［(53.37±8.92)% vs (53.51±5.84)%, P&gt;0.05］. But 6 months later, LVEF in the BMCs group were clearly higher than that in the control group ［(59.33±12.91)% vs (50.30±8.30)%, P&lt;0.05］. (3)There were no evident difference in EDV or ESV between two groups at baseline ［EDV:(113.74±23.24) ml vs (129.94±32.72) ml , P&gt;0.05; ESV: (57.12±18.66) ml vs (62.09±17.68) ml, P&gt;0.05］. Three months later, EDV and ESV in the control group were markedly greater than those in the BMCs group ［EDV: (154.89±46.34) ml vs (104.85±33.21) ml, P&lt;0.05; ESV :(82.91±35.79) ml vs (49.54±23.32) ml, P&lt;0.05］. But EDV and ESV did not change much from 3rd to 6th month in either group (P&gt;0.05). Conclusions Emergency transplantation of autologous BMCs in patients with acute myocardial infarction helps to improve global and regional contractility and attenuate post-infarction left ventricular remodeling. Tissue tracking and strain imaging provide quick, simple and noninvasive methods for quantifying left ventricular segmental function in humans.     

自体骨髓干细胞移植治疗心肌梗死的临床研究

目的研究自体骨髓干细胞经冠脉移植对心肌梗死患者心功能的影响及其安全性。方法6例冠心病伴心肌梗死患者,男5例、女1例,平均年龄62岁(51~75岁),体外分离培养自体骨髓干细胞2~3周,开通梗死相关“罪犯”血管,通过冠脉转运将骨髓干细胞植入患者的心肌梗死区,移植的骨髓间充质干细胞数为(0.9~3.2)×106个,骨髓单个核细胞数为(1.6~5.3)×106个。术前和术后6个月分别行99mTc-MIBI心肌灌注显像、二维超声心动图、心电图和动态心电图检查。结果6例患者二维超声心动图检查示左室射血分数(LVEF)较术前明显增加[(52.33±8.70)%vs(42.92±12.60)%,P<0.05],左室舒张期内径(LVDd)较术前明显减少[(54.33±4.84)vs(61.67±6.83),P<0.05];6例99mTc-MIBI检查显示梗死部位心肌灌注明显改善。术中及术后随访6~18个月无心律失常和其他合并症发生。结论进行自体骨髓干细胞经冠脉移植治疗冠心病、心肌梗死具有抑制左室重构、改善心脏功能的作用。     

大鼠骨髓单个核细胞移植治疗心肌梗死

目的:探讨大鼠骨髓单个核细胞(mononuclear bone marrow cells,MBMCs)移植方法对梗死心肌的治疗效果.方法:无菌条件下取成年雄性近交系Wistar大鼠股骨和胫骨,常规抽取骨髓,分离制备MBMCs悬液.将成年雄性近交系Wistar大鼠46只随机分为空白对照组(组Ⅰ,n=10)、梗死对照组(组Ⅱ,n=18)、细胞移植组(组Ⅲ,n=18).用结扎冠状动脉左前降支的方法建立大鼠心肌梗死模型,在建模后2周分别将10μl MBMCs(6×106)悬液和等量磷酸盐缓冲液(PBS)植入组Ⅱ和组Ⅲ的梗死周边区.用心脏超声观察细胞移植后4周的心脏功能改变,用免疫荧光和免疫组化及电镜观察移植细胞在心肌梗死区及梗死周边区的存活、增殖和分化情况.结果:MBMCs移植4周后,移植组超声检查左室功能重建指标如左室射血分数(EF)、左室短轴缩短率(FS)、左室前壁厚度(LVAWT)均较梗死对照组改善明显(P＜0.05);移植组血流动力学检查指标如左室收缩压(LVSP)、左室舒张末压(LVEDP)和左室舒张压力时间变化最大速率(±dp/dtmax)也优于梗死对照组(P＜0.05).MBMCs移植6周后,大鼠心肌梗死区有移植细胞存活、增殖,并分化为有心肌细胞特征的细胞.结论:实验证实用MBMCs移植治疗心肌梗死,可以明显改善梗死后心脏功能.这一效果可能与MBMCs移植相关的心肌再生和新生血管形成有关.     

曲美他嗪联合骨髓间质干细胞移植治疗急性心肌梗死的实验研究

目的探讨曲美他嗪(TMZ)能否改善骨髓间质干细胞(MSCs)在体外缺氧模型及急性心肌梗死(AMI)大鼠心脏的存活。方法加入或未加入TMZ的MSCs在无血清培养基培养并缺氧暴露12h,采用透射电子显微镜和流式细胞仪检查第3代MSCs的活力和凋亡。30只Wistar大鼠随机分为AMI对照组、MSCs组及(MSCs+TMZ)组,结扎左冠状动脉前降支制备AMI模型。将MSCs注入梗死心肌边缘[(MSCs组和(MSCs+TMZ)组)]。(MSCs+TMZ)组的大鼠在AMI前3d开始至AMI后28d加喂TMZ。移植28d后,超声心动图评估心脏结构和功能。免疫荧光染色检测移植细胞在体内的存活和分化。TUNEL法检测细胞凋亡。收集TMZ治疗开始前和AMI后24、48h的血液样本,测量C反应蛋白(CRP)与肿瘤坏死因子-α(TNF-α)的变化。结果缺氧培养下,TMZ处理过的MSCs细胞凋亡降低了一半。在体内与AMI对照组相比,MSCs组和(MSCs+TMZ)组的心肌梗死面积显著缩小,心功能明显改善。与单纯MSCs移植相比,TMZ与MSCs移植的组合治疗表现出了更低的干细胞凋亡、更高的干细胞存活、更小的心肌梗死面积和进一步改善的心功能。各组之间CRP、TNF-α的基线水平并无显著差异,然而24h时(MSCs+TMZ)组的所有参数均低于MSCs组。结论 MSCs移植添加TMZ治疗AMI增加MSCs存活和心脏功能的恢复上优于单纯MSCs移植,抑制炎症因子表达可能是其机制之一。     

经心外膜向心肌移植自体骨髓单核细胞治疗急性心肌梗死的实验研究

目的 在小型猪动物模型上探讨自体骨髓单核细胞移植到心肌梗死局部对心功能和梗死心肌代谢的影响.方法 结扎小型猪冠状动脉前降支建立急性心肌梗死动物模型.分为3组:空白对照组、心肌梗死1周后处理组及心肌梗死2周后处理组,处理组经心外膜向心肌注射自体骨髓单核细胞.然后检测各项指标.结果 空白对照组与处理组心尖室间隔下部室壁增厚率差异具有统计学意义.显影细胞区域冰冻切片免疫组化测定,证实自体骨髓单核细胞分化细胞转化为心肌或内皮细胞并证实其存活增殖功能.结论 经心外膜向梗死区域的心肌移植自体骨髓单个核细胞,可以改善梗死区域心肌的收缩功能.移植的单个核细胞可以在梗死区域中存活,增殖,分化和迁移,增加梗死区域的微血管数量,再生、再造心肌.     

犬自体骨髓单个核细胞经冠状动脉移植重建梗死心肌

目的:探讨选用含有原始干细胞/多系祖细胞及基质细胞的骨髓单个核细胞(MBMC),经冠状动脉移植重建犬梗死心肌,观察其修复、重建梗死心肌及改善心功能的效果.方法:经皮穿刺球囊封堵前降支成功建立心肌梗死动物模型并存活约1周的杂种犬10条,平均分为细胞移植组和对照组.细胞移植术前1 d分离MBMC,以DAPI标记,温箱保存过夜.常规冠状动脉造影、PTCA,在球囊闭塞前降支条件下注入MBMC(每次4×106个细胞,2 ml),气囊保持充盈2 min后恢复灌注2 min,重复注入3次.分别于术前及术后2个月行超声心动图检查,动物放血处死后取心脏前壁和下壁冰冻切片,荧光显微镜观察并照相.结果:移植组术后与术前及对照组比较,平均左室舒张末容量和左室收缩末容量减少、EF和FS增加(P＜0.05);对照组则左室舒张末容量和左室收缩末容量明显增加、FS减少(P＜0.05).细胞移植组左室收缩及舒张内径和梗死长度均较移植前缩小,梗死部位室壁厚度较移植前增加(P＜0.05);而对照组梗死区室壁变薄.在细胞移植组前壁心肌组织冰冻切片中可以看到心肌细胞核发黄绿色荧光,发现带绿色荧光的丝状肌丝肌管,移植犬的下壁和对照组的前壁可见心肌细胞核发出光亮较移植组前壁淡的自发黄绿色荧光,而均未发现有带绿色荧光的丝状肌丝肌管.结论:自体骨髓单个核细胞移植可重建坏死心肌,梗死区域内有移植的新生心肌细胞,且抑制心室的重塑、防止心脏进一步扩大和心功能下降.     

自体骨髓干细胞动员治疗心肌梗死

目的探讨基因重组人粒细胞集落刺激因子(rhGCSF)动员骨髓干细胞对急性心肌梗死动物模型的治疗作用。方法将60只SD大鼠随机分成对照组和治疗组,结扎冠状动脉左前降支建立大鼠心肌梗死实验模型。治疗组在复制大鼠心肌梗死模型3h后用经生理盐水稀释的rhGCSF(2mg·L-1)皮下注射10μg·kg-1·d-1,共5d;对照组复制大鼠心肌梗死模型3h后皮下注射等量生理盐水,共5d。在复制大鼠心肌梗死模型24、48h和2周后,两组分别先后各取10只大鼠行在体心功能(±dpdtmax)测定,测定完毕后各组随机选取6只大鼠处死行心脏梗死范围的测量,其余大鼠处死取出心脏行HE染色及CD34+免疫组织化学分析。结果结扎冠状动脉左前降支24、48h和2周后治疗组±dpdtmax均高于对照组(P<0.01);治疗组心脏梗死范围低于对照组(P<0.01)。病理检查显示对照组心肌梗死区周围有较多以中性粒细胞为主的炎症细胞浸润,而治疗组大鼠心肌坏死程度较对照组轻,浸润的细胞以CD34+的单个核细胞为主,并可见新生的心肌细胞生长。结论rhGCSF动员自体骨髓干细胞原位移植可修复梗死心肌、减少梗死范围、提高心功能,可用于急性心肌梗死的治疗。     

加味四物汤动员心肌梗死小鼠骨髓干细胞改善心室重构

目的 观察加味四物汤(mSWT)对心肌梗死(MI)小鼠骨髓干细胞动员的影响,探讨mSWT改善心室重构的机制.方法 结扎冠状动脉前降支建立小鼠MI模型,灌胃给予mSWT,小鼠被分为4组:假手术对照组,MI模型组,mSWT低剂量(5 g·kg-1·d-1)组和mSWT高剂量(10 g·kg-1·d-1)组,每组10只.行心肌组织病理学检查,流式细胞术分析造血干细胞(HSCs)和间充质干细胞(MSCs)占骨髓和外周血细胞的百分比.结果 mSWT高、低剂量组治疗14 d后,较MI模型组小鼠左心室壁较厚,MI面积减小,梗死区域可见大量岛状分布的心肌细胞,心室重构明显改善.且小鼠骨髓和外周血中HSCs及MSCs细胞百分比明显增加(P＜0.05).结论 mSWT可能通过促进MI小鼠骨髓HSCs和MSCs的增殖和向外周动员,参与改善MI后的心室重构.     

兔骨髓间充质干细胞的筛选与体外培养

目的 :探讨兔骨髓间充质干细胞 (mesenchymalstemcells ,MSCs)体外分离筛选及扩增的条件 ,同时观察骨髓细胞体外培养的生物学特性。方法 :无菌条件下采集兔骨髓 ,肝素抗凝 ,经淋巴细胞分层液 (相对密度为 1 .0 77)密度梯度离心分离骨髓单个核细胞 ,进而贴壁培养 ,获得MSCs。采用常用细胞培养技术和细胞培养的研究方法 ,观察不同贴壁时间、不同种植密度、有无包被成分及不同蛋白包被培养板对MSCs生长增殖的影响。结果 :预先蛋白包被培养板有利于MSCs贴壁。在细胞生长和增殖方面 ,纤粘连蛋白优于明胶和Ⅰ型胶原 ,以贴壁 48～ 72h ,种植密度 (3～ 9)× 1 0 4 /ml为最适条件 ,有利于梭形细胞形态和快速增殖能力的维持 ,保持成体干细胞的多向分化性。结论 :建立了MSCs体外分离和培养的最适条件 ,为进一步研究MSCs的诱导分化和应用提供了保证     

超声微泡辅助干细胞移植治疗急性心肌梗死的实验研究

目的:探讨超声微泡在自体骨髓间充质干细胞(BMMSCs)移植治疗猪急性心肌梗死(AMI)改善心功能中的作用。方法:选取14只3~4月龄小型家猪,抽取骨髓,分离、培养、标记BMMSCs,球囊封堵猪左冠状动脉前降支(LAD)建立AMI模型,分为实验组(超声微泡作用下移植干细胞,n=8)和对照组(无超声微泡作用,n=6),7 d后向LAD梗死处输注自体BMMSCs移植治疗。在移植前1 d和移植后6周分别行64层螺旋CT心脏扫描及组织病理学检查。结果:与对照组相比,实验组心功能改善更为明显,梗死区毛细血管数(16.37±3.20vs9.50±1.87,P<0.05)、普鲁士蓝染色阳性细胞数(71.63±14.09vs40.67±9.48,P<0.05)、结蛋白免疫组化染色阳性细胞数(31.88±4.16vs19.00±2.28,P<0.05)均明显增加。结论:超声微泡增效自体BMMSCs移植治疗AMI,能更有效地改善心肌梗死后的心功能,促进血管新生。     

MMSCs移植对大鼠心肌梗死后MMP-2,TIMP-2表达及心室重构的影响

目的:研究骨髓间充质干细胞(MMSCs)移植对实验性心肌梗死(MI)大鼠心肌细胞外基质金属蛋白酶(MMP-2)和基质金属蛋白酶抑制剂(TIMP-2)的表达及心室重构的影响.方法:采用开胸结扎冠状动脉左前降支(LAD)的方法制作MI大鼠模型,造模成功后12～14d经大鼠舌下静脉移植已转染增强型绿色荧光蛋白(pEGFP-C1)的MMSCs,分4,8wk2个时段检测大鼠左心及全心质量指数、胶原含量及MMP-2,TIMP-2的量效变化,荧光显微镜观察绿色荧光的表达.结果:4wk时移植组左心质量指数为[(1.69×10-3)±(11.02×10-5)],全心质量指数为[(2.22×10-3)±(5.08×10-5)]均较同期模型组[(1.83×10-3)±(9.48×10-5)],[(2.47×10-3)±(17.16×10-5]显著降低(P<0.01).8wk时其变化趋势与4wk时相同.4wk时移植组的胶原含量为(11.19±1.15)%较同期模型组的胶原含量(27.89±5.59)%显著降低(P<0.01),8wk时变化趋势与4wk时相同.移植组MMP-2的表达显著低于同期模型组中MMP-2的表达,而TIMP-2的...     

骨髓单个核细胞移植对急性心梗大鼠血流动力学的影响

目的:评价局部移植骨髓单个核细胞对急性心梗大鼠缺血心肌血管生成作用及血流动力学的影响.方法:将26只Lweis大鼠随机分为移植组(n=14)及对照组(n=12),结扎大鼠左冠状动脉,建立急性心肌梗死模型.将新分离的骨髓单个核细胞注射到大鼠的缺血心肌中(对照组大鼠注射等量无菌PBS),术后4周观察心肌血管数目,结合心脏血流动力学参数评价心功能的改善情况.结果:4周后移植组与对照组相比左心室舒张末期压力明显降低(P＜0.01),压力变化率最大值(dp/dt)明显升高(P＜0.01,P＜0.05).移植组心肌梗死区有明显血管增生,移植组和对照组心肌血管密度分别为(23.1±1.5)和(10.5±1.8)个/HPF,差异有显著性意义(P＜0.01).结论:局部移植骨髓单个核细胞能明显促进缺血心肌新生血管生成,增加缺血区灌注,并可在一定程度上改善其血流动力学指标.     

自体骨髓干细胞移植治疗心肌梗死患者心肌代谢及收缩功能的变化

目的:探讨自体骨髓间充质干细胞(BMSCs)和单个核细胞(BMMNCs)经梗死相关冠状动脉(IRA)移植对冠心病心肌梗死患者近、中期心功能、心肌代谢的影响及其安全性。方法:对14例冠心病伴心肌梗死史1~35个月的患者。细胞移植组8例,通过冠脉系统将BMSCs和BMMNCs植入心肌梗死区,余6例常规治疗。两组患者均于术前和术后3,8个月行SPECT双核素心肌显像、二维超声心动图、6 m in步行试验及动态心电图检查。结果:移植组患者在梗死相关冠脉开通后注入干细胞,8例心功能从Ⅱ~Ⅳ级提高至Ⅰ~Ⅱ级,6 m in步行试验从(320.72±60.47)m提高至(406.88±68.31)m,6个月后又提高至(484.79±81.00)m。6例前壁心梗患者3,8月后LVEF由术前的42.5%提高至49.5%和54%;术后3,8月心肌双核素检查缺损区占整个心肌体积由术前的45.5%减少至39%和25%。6例对照组患者术后3月EF较术前提高,心肌双核素检查缺损区占心肌体积减少,术后8月与术后3月无明显改变。2例下壁心肌梗死患者经干细胞移植临床效果不明显。结论:自体BMSCs和BMMNCs经冠脉移植治疗冠心病心肌梗死,可以显著改善前壁心肌梗死患者左室收缩功能和心肌代谢。     

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.

     

双特异抗体增效骨髓间充质干细胞干预心肌纤维化

目的:探讨在双特异抗体(Bispecific antibody,BiAb)的辅助下,骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)移植干预心肌纤维化的效果.方法:以anti-CD29(能识别间充质干细胞)和抗肌凝蛋白轻链抗体(Anti-myosin light chain antibody,AMLCA)(能特异性结合损伤心肌)制备BiAb(CD29×AMLCA).将此BiAb与雄性小鼠BMSCs经尾静脉输入异丙肾性心肌纤维化雌性小鼠(BMSCs+BiAb组).另设单纯BMSCs组、单纯BiAb组、正常对照组、未治疗组.5周后处死小鼠,荧光定量PCR检测心肌y染色体鉴别基因(Sex-determining region of Y-chromosome,SRY)、基质金属蛋白酶-9(Matrix metalloproteinases-9,MMP-9)、基质金属蛋白酶组织抑制剂-1(Tissue inhibitor of metalloproteinase-1,TIMP-1)的表达.天狼猩红染色比较心脏胶原含量.结果:BMSCs+BiAb组的干细胞归巢数比BMSCs组多(P＜0.05).与未治疗组相比,BMSCs+BiAb组和单纯BMCSs组的心肌MMP-9和TIMP-1表达下调,心脏胶原沉积降低(P＜0.05).与BMSCs组相比,BMSCs+BiAb组的心肌纤维化改善更为明显(P＜0.05).结论:CD29×AMLCA双特异抗体可促进小鼠骨髓间充质干细胞的归巢.BMSCs归巢后能改善缺血心肌MMP-TIMP表达,干预心肌纤维化.     

自体骨髓单个核细胞植入对心肌梗死后心脏功能的影响

 目的 探讨骨髓单个核细胞植入对心肌梗死后心脏功能的影响。 方法 以冷冻损伤的方法建立新西兰种家兔心肌梗死模型,分为骨髓单个核细胞组（mononuclear bone marrow cells,MN-BMCs组）和对照组,各8只。4周后, MN-BMCs组中将提取分离的MN-BMCs,植入心肌梗死及其边缘区,对照组于心肌梗死及其边缘区注射相应体积的IMDM完全培养液。以超声心动图分析细胞植入前后心脏功能改善情况。 结果 MN-BMCs植入心肌梗死及其边缘区可以改善局部及整体心脏功能：细胞移植4周后两组之间比较,MN-BMCs组局部心脏功能明显优于对照组,表现为MN-BMCs组比对照组心肌梗死区室壁厚度增加,（4.81±0.55）mm vs 3.94±0.54）mm,P＜0.05;收缩期和舒张期心肌梗死区室壁运动速度明显增加,分别为（8.20±0.77）cm/s vs 7.00±0.56）cm/s,P＜0.05,（8.51±0.93） cm/s vs （6.75 ±0.57）cm/s,P＜0.05。细胞移植4周后,MN-BMCs组整体心脏功能亦优于对照组,表现为MN-BMCs组比对照组心腔减小,LVDD分别为 （14.89±1.36）mm vs （16.24±1.31）mm,P＜0.05,LVSD分别为（10.48±1.32）mm vs （12.29±1.48）mm,P＜0.05;MN-BMCs组左心室收缩功能明显改善,左室射血分数及缩短分数提高,LVEF分别为（65.25±5.73）% vs （57.00±5.81）%,P＜0.05,LVFS分别为 （30.25±3.73）% vs （24.88±3.95）%,P＜0.05。 结论 骨髓单个核细胞植入心肌梗死及其边缘区可以改善心肌梗死后心脏功能。     

PC-3细胞中雄激素受体突变体的表达和转录激活功能的研究

目的:探讨不同时间点移植骨髓单个核细胞(bone marrow mononuclear cells,BMMCs)对心肌梗死后的心功能的影响.方法:取成年雄性近交系Wistar大鼠股骨和胫骨,常规抽取骨髓,分离制备BMMCs悬液.将成年雄性近交系Wistar大鼠64只随机分为空白对照组(组Ⅰ,n=10)、梗死对照组(组Ⅱ,n=18)、即刻移植组(组Ⅲ,n=18)和两周移植组(组Ⅳ,n=18).用结扎冠状动脉左前降支的方法建立大鼠心肌梗死模型,在建模后即刻和2周分别将10μl BMMCs液(6×106个细胞)植入组Ⅲ和组Ⅳ的梗死周边区,组Ⅱ于建模后14 d植入等量的磷酸盐缓冲液(PBS).用心脏超声和血流动力学检查观察细胞移植后4周的心脏功能改变.结果:骨髓单个核细胞移植4周后,与组Ⅱ、组Ⅲ相比,组Ⅳ左室功能重建指标[左室射血分数(EF)、左室短轴缩短率(FS)、左室前壁厚度(LVAWT)]和血流动力学检查指标[左室收缩压(LVSP)、左室舒张末压(LVEDP)和左室舒张压力时间变化最大速率±dp/dtmax]均明显改善(P＜0.05);而组Ⅱ、Ⅲ间上述指标相比无显著差异.结论:心肌梗死后2周骨髓单个核细胞移植治疗效果优于即刻移植,可能是由于在心肌梗死后1周时炎症反应最重,而2周时逐渐减弱.