背景:骨髓干细胞移植可改善心功能、预防心室重构,目前用于移植的成体骨髓干细胞有骨髓单个核细胞、骨髓间充质干细胞和内皮祖细胞等,不间种类的骨髓干细胞移植后的效果及具体机制尚不清楚.目的:比较自体骨髓单个核细胞、骨髓间充质干细胞冠状动脉移植对急性心肌梗死后心室重构的影响.设计、时间及地点:随机对照动物实验,于2005-03/2006-12在河北省人民医院临床研究中心、河北医科大学电镜室及大连宝生物进行.(basic fibroblast growth factor,bFGF)材料:36只冀中白猪随机分为4组:假手术组6只、模型组10只、骨髓单个核细胞组10只、骨髓间充质干细胞组10只.方法:采用梯度密度离心法分离培养猪自体骨髓单个核细胞,以贴壁法分离培养猪自体骨髓间充质干细胞,移植前均行胶体会标记.除假手术组外,其余3组均以球囊导管压迫冠状动脉前降支的方法建立猪急性心肌梗死模型.造模后90min,经导管由冠状动脉腔内骨髓单个核细胞组移植自体骨髓单个核细胞(6.0±1.3)×107个,骨髓间充质干细胞组移植自体骨髓间充质干细胞(4.5±2.1)×107个,培养28d.主要观察指标:光镜及电镜观察心肌组织病理学改变:超声检查心功能变化:免疫组织化学检测心肌血管数、心肌细胞凋亡、心肌组织核凼子κB及肌钙蛋白Ⅰ阳性表达情况,及其与心功能的关系:RT-PCR法检测心肌血管内皮生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGE)mRNA的表达,及其与心功能的关系.结果:①骨髓间充质干细胞组在梗死区、梗死边缘区均可见大量血管增生,冠脉血管周围可见异常细胞团生长,骨髓单个核细胞组有较多的毛细血管"芽生"现象.②细胞移植前各组心功能指标基本相似(F=1.550,P＞0.05).移植后28 d与模型组比较,其余3组左心室射血分数均明显降低(F=5.30,P＜0.05).③与模型组比较,骨髓单个核细胞组梗死区及梗死边缘区的血管数明显增加(F=29.56～34.87,P＜0.01),骨髓间充质干细胞组无变化;移植细胞两组的梗死区、梗死边缘区心肌细胞凋亡率均显著降低(F=14.31～35.34,P＜0.01),肌钙蛋白Ⅰ阳性率均明显升高(F=19.05,P＜0.01);梗死边缘区核因子κB阳性率均明显降低(F=19.05,P＜0.01).④骨髓单个核细胞组梗死边缘区VEGF基因的表达显著高于模型组、骨髓间充质千细胞组(F=49.41,P＜0.01).骨髓问充质干细胞组梗死边缘区bFGF基因的表达显著高于模型组、骨髓单个核细胞组(F=4.71,P＜0.01).⑤左室射血分数与心肌细胞凋亡率、心肌核因子κB呈负相关(r=-0.4411,P＜0.05;r=-0.5796,P＜0.01);与血管数、VEGF及bFGF表达呈正相关(r=0.775,P＜0.01;r=0.5651,P＜0.05;r=0.573 5,P＜0.05).结论:经冠脉自体骨髓单个核细胞或骨髓间充质干细咆移植均可减轻心肌梗死后左心室重构,心功能的改善与干细胞移植后增加心肌血管数量及心肌VEGF,bFGF表达、减少心肌细胞凋亡及核因子κB水平有关.骨髓单个核细胞移植促心肌血管增生及.VEGF的表达均优于骨髓间充质干细胞,而后者促bFGF基因表达的作用优于前者.
揹景:骨髓榦細胞移植可改善心功能、預防心室重構,目前用于移植的成體骨髓榦細胞有骨髓單箇覈細胞、骨髓間充質榦細胞和內皮祖細胞等,不間種類的骨髓榦細胞移植後的效果及具體機製尚不清楚.目的:比較自體骨髓單箇覈細胞、骨髓間充質榦細胞冠狀動脈移植對急性心肌梗死後心室重構的影響.設計、時間及地點:隨機對照動物實驗,于2005-03/2006-12在河北省人民醫院臨床研究中心、河北醫科大學電鏡室及大連寶生物進行.(basic fibroblast growth factor,bFGF)材料:36隻冀中白豬隨機分為4組:假手術組6隻、模型組10隻、骨髓單箇覈細胞組10隻、骨髓間充質榦細胞組10隻.方法:採用梯度密度離心法分離培養豬自體骨髓單箇覈細胞,以貼壁法分離培養豬自體骨髓間充質榦細胞,移植前均行膠體會標記.除假手術組外,其餘3組均以毬囊導管壓迫冠狀動脈前降支的方法建立豬急性心肌梗死模型.造模後90min,經導管由冠狀動脈腔內骨髓單箇覈細胞組移植自體骨髓單箇覈細胞(6.0±1.3)×107箇,骨髓間充質榦細胞組移植自體骨髓間充質榦細胞(4.5±2.1)×107箇,培養28d.主要觀察指標:光鏡及電鏡觀察心肌組織病理學改變:超聲檢查心功能變化:免疫組織化學檢測心肌血管數、心肌細胞凋亡、心肌組織覈凼子κB及肌鈣蛋白Ⅰ暘性錶達情況,及其與心功能的關繫:RT-PCR法檢測心肌血管內皮生長因子(vascular endothelial growth factor,VEGF)、堿性成纖維細胞生長因子(basic fibroblast growth factor,bFGE)mRNA的錶達,及其與心功能的關繫.結果:①骨髓間充質榦細胞組在梗死區、梗死邊緣區均可見大量血管增生,冠脈血管週圍可見異常細胞糰生長,骨髓單箇覈細胞組有較多的毛細血管"芽生"現象.②細胞移植前各組心功能指標基本相似(F=1.550,P＞0.05).移植後28 d與模型組比較,其餘3組左心室射血分數均明顯降低(F=5.30,P＜0.05).③與模型組比較,骨髓單箇覈細胞組梗死區及梗死邊緣區的血管數明顯增加(F=29.56～34.87,P＜0.01),骨髓間充質榦細胞組無變化;移植細胞兩組的梗死區、梗死邊緣區心肌細胞凋亡率均顯著降低(F=14.31～35.34,P＜0.01),肌鈣蛋白Ⅰ暘性率均明顯升高(F=19.05,P＜0.01);梗死邊緣區覈因子κB暘性率均明顯降低(F=19.05,P＜0.01).④骨髓單箇覈細胞組梗死邊緣區VEGF基因的錶達顯著高于模型組、骨髓間充質韆細胞組(F=49.41,P＜0.01).骨髓問充質榦細胞組梗死邊緣區bFGF基因的錶達顯著高于模型組、骨髓單箇覈細胞組(F=4.71,P＜0.01).⑤左室射血分數與心肌細胞凋亡率、心肌覈因子κB呈負相關(r=-0.4411,P＜0.05;r=-0.5796,P＜0.01);與血管數、VEGF及bFGF錶達呈正相關(r=0.775,P＜0.01;r=0.5651,P＜0.05;r=0.573 5,P＜0.05).結論:經冠脈自體骨髓單箇覈細胞或骨髓間充質榦細咆移植均可減輕心肌梗死後左心室重構,心功能的改善與榦細胞移植後增加心肌血管數量及心肌VEGF,bFGF錶達、減少心肌細胞凋亡及覈因子κB水平有關.骨髓單箇覈細胞移植促心肌血管增生及.VEGF的錶達均優于骨髓間充質榦細胞,而後者促bFGF基因錶達的作用優于前者.
배경:골수간세포이식가개선심공능、예방심실중구,목전용우이식적성체골수간세포유골수단개핵세포、골수간충질간세포화내피조세포등,불간충류적골수간세포이식후적효과급구체궤제상불청초.목적:비교자체골수단개핵세포、골수간충질간세포관상동맥이식대급성심기경사후심실중구적영향.설계、시간급지점:수궤대조동물실험,우2005-03/2006-12재하북성인민의원림상연구중심、하북의과대학전경실급대련보생물진행.(basic fibroblast growth factor,bFGF)재료:36지기중백저수궤분위4조:가수술조6지、모형조10지、골수단개핵세포조10지、골수간충질간세포조10지.방법:채용제도밀도리심법분리배양저자체골수단개핵세포,이첩벽법분리배양저자체골수간충질간세포,이식전균행효체회표기.제가수술조외,기여3조균이구낭도관압박관상동맥전강지적방법건립저급성심기경사모형.조모후90min,경도관유관상동맥강내골수단개핵세포조이식자체골수단개핵세포(6.0±1.3)×107개,골수간충질간세포조이식자체골수간충질간세포(4.5±2.1)×107개,배양28d.주요관찰지표:광경급전경관찰심기조직병이학개변:초성검사심공능변화:면역조직화학검측심기혈관수、심기세포조망、심기조직핵당자κB급기개단백Ⅰ양성표체정황,급기여심공능적관계:RT-PCR법검측심기혈관내피생장인자(vascular endothelial growth factor,VEGF)、감성성섬유세포생장인자(basic fibroblast growth factor,bFGE)mRNA적표체,급기여심공능적관계.결과:①골수간충질간세포조재경사구、경사변연구균가견대량혈관증생,관맥혈관주위가견이상세포단생장,골수단개핵세포조유교다적모세혈관"아생"현상.②세포이식전각조심공능지표기본상사(F=1.550,P＞0.05).이식후28 d여모형조비교,기여3조좌심실사혈분수균명현강저(F=5.30,P＜0.05).③여모형조비교,골수단개핵세포조경사구급경사변연구적혈관수명현증가(F=29.56～34.87,P＜0.01),골수간충질간세포조무변화;이식세포량조적경사구、경사변연구심기세포조망솔균현저강저(F=14.31～35.34,P＜0.01),기개단백Ⅰ양성솔균명현승고(F=19.05,P＜0.01);경사변연구핵인자κB양성솔균명현강저(F=19.05,P＜0.01).④골수단개핵세포조경사변연구VEGF기인적표체현저고우모형조、골수간충질천세포조(F=49.41,P＜0.01).골수문충질간세포조경사변연구bFGF기인적표체현저고우모형조、골수단개핵세포조(F=4.71,P＜0.01).⑤좌실사혈분수여심기세포조망솔、심기핵인자κB정부상관(r=-0.4411,P＜0.05;r=-0.5796,P＜0.01);여혈관수、VEGF급bFGF표체정정상관(r=0.775,P＜0.01;r=0.5651,P＜0.05;r=0.573 5,P＜0.05).결론:경관맥자체골수단개핵세포혹골수간충질간세포이식균가감경심기경사후좌심실중구,심공능적개선여간세포이식후증가심기혈관수량급심기VEGF,bFGF표체、감소심기세포조망급핵인자κB수평유관.골수단개핵세포이식촉심기혈관증생급.VEGF적표체균우우골수간충질간세포,이후자촉bFGF기인표체적작용우우전자.
BACKGROUND:Bone marrow stem cell transplantation can improve heart function and prevent ventricle remodeling.At present,the adult bone marrow stem cells used for transplantation primarily included bone marrow mononuclear cells (BM-MNCs) and mesenchymal stem cells (MSCs),and endothelial progenitor cells.The curative effects and precise mechanisms of transplantation of various bone marrow stem cells remain unknown.OBJECTIVE:To compare the effects of transplantation of autologous BM-MNCs and MSCs via the coronary artery on ventricle remodeling subsequent to acute myocardial infarction (AMI). DESIGN,TIME AND SETTING:Randomized controlled animal experiment performed at the Center for Clinical Research,Hebei Provincial People's Hospital,Electron Microscope Room,Hebei Medical University between March 2005 and December 2006.MATERIALS:Thirty-six male Jizhong pigs,were randomly divided into 4 groups:control group (n = 6),infarct model group (n = 10),BM-MNC group (n = 10),and MSC group (n = 10).METHODS:Porcine autologous BM-MNCs were isolated by gradient density centrifugation,and MSCs were obtained by adherence method.Prior to transplantation,both BM-MNCs and MSCs were colloidal gold labeled.Except the infract model group,pigs in the other 3 groups were developed into AMI models by oppressing the left anterior descending branch with balloon catheter.Ninety minutes after modeling,(6.0±1.3)×107 autologous BM-MNCs and (4.5±2.1)x 107 MSCs were respectively transplanted into pigs in the BM-MNC group and the MSC group via the coronary artery and cultured for 28 days.MAIN OUTCOME MEASURES:Observation of pathological changes of cardiac muscle tissue by light and electron microscope;Examination of cardiac function by ultrasonograph;Detection of the number of blood vessels and apoptotic myocardial cells,and expression of nuclear factor-κB (NF-κB) and troponin Ⅰ and its correlation to cardiac function by immunohistochemistry;Detection of mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in the cardiac tissue as well as its correlation to cardiac function by reverse transcription-polymerase chain reaction (RT-PCR).RESULTS:In the MSC group,there was proliferation of a great deal of blood vessels as well as growth of abnormal cell masses around the coronary vessels,while the BM-MNC group exhibited the "budding" of many capillary vessels.Prior to transplantation,cardiac function indices were basically similar among each group (F = 1.550,P＞0.05).Twenty-eight days after transplantation,left ventricular ejection fraction was significantly lower in the control,BM-MNC,and MSC groups than in the infarct model group (F = 5.30,P＜0.05),while endocardial fractional shortening was significantly higher (F = 10.67,P＜0.01).Compared with the infarct model group,the number of blood vessels in the infarct zone and infarct border zone was increased in the BM-MNC group (F=29.56-34.87,P＜0.01) and had no apparent change in the MSC group.In the BM-MNC and MSC groups,apoptotic myocardial cells in the infarct zone and infarct border zone were significantly reduced (F=14.31-35.34,P＜0.01 ) and troponin I expression rate was significantly increased (F=19.05,P＜0.01 ),as compared with the infarct model group.In addition,NF-κB positive rate in the infarct border zone was significantly lower in the BM-MNC and MSC groups than in the infarct model group (F=19.05,P＜0.01).VEGF gene expression level in the infarct border zone was significandy higher in the BM-MNC group than in the infarct model group and MSC group (F = 49.41,P＜0.01).bFGF gene expression level in the infarct border zone was significantly higher in the MSC group than in the infarct model and BM-MNC groups (F=4.71,P＜0.01).LVEF was negatively correlated to myocardial cell apoptosis rate and NF-κB level (r=-0.441 1,P＜0.05;r=-0.579 6,P＜0.01 ).LVEF was positively correlated to number of blood vessels,VEGF and bFGF expression (r=0.775,P＜0.01;r=0.565 1,P＜0.05;r=0.573 5,P＜0.05).CONCLUSION:Transplantation of both autologous BM-MNC and MSC via coronary artery can improve the condition of left ventricular remodeling subsequent to myocardial infarction.The improvement of cardiac functions is related to the increase of blood vessels,VEGF and bFGF expression,the decrease of myocardial cell apoptosis and NF-κ B level in cardiac muscle tissues after stem cell transplantation.BM-MNC transplantation better promotes blood vessel proliferation and VEGF expression in the cardiac tissue but produces worse effects on bFGF gene expression than MSC transplantation.
