CAJ | 학술논문

背景:骨髓间质干细胞和血管内皮细胞生长因子移植具有促进血管再生、改善心功能的作用,但是二者联合应用是否优于单独应用尚不清楚.目的:观察同种异体骨髓间质干细胞移植联合血管内皮生长因子基因转染对大鼠急性心肌梗死血管再生和心功能的影响.设计:大鼠骨髓间质干细胞培养采用单一样本观察;细胞移植和基因转染采用随机对照动物实验.单位:华中科技大学协和医院心内科,同济医学院心血管病研究所.材料:健康雄性Wistar大鼠94只.表达载体PAdTrack/VEGF165.方法:实验于2004-06/2005-06在同济医学院心血管病研究所实验室完成.①体外分离、纯化、培养大鼠骨髓间质干细胞,以5-溴-2'-脱氧尿苷标记细胞.②制备、抽提、纯化、鉴定质粒PAdTrack/VEGF165.③结扎冠状动脉建立急性心肌梗死模型2周后,随机将其分为4组,每组均为12只,干细胞+质粒组、干细胞组、质粒组、对照组,分别进行心肌内大鼠骨髓间质干细胞移植和/或VEGF165转染、DMEM注射.④4周后行免疫组织化学和超声心动图检查.主要观察指标:①各组大鼠梗死及缺血区免疫组织化学和苏木精-伊红染色检查.②血管计数.③超声心动图检查.结果:48只大鼠进入结果分析.①干细胞+质粒组和干细胞组梗死及缺血心肌处可见大量5-溴-2'-脱氧尿苷标记的移植细胞,其中缺血心肌处部分移植细胞分化为血管内皮细胞并形成新生毛细血管.②Ⅷ因子染色阳性的新生血管密度分布为干细胞+质粒组＞质粒组＞干细胞组＞对照组(P均＜0.01).③细胞移植和基因转染治疗后室壁厚度和室壁运动幅度改善,射血分数值增加幅度为干细胞+质粒组＞干细胞组＞质粒组＞对照组(P均＜0.01).结论:同种异体骨髓间质干细胞移植联合血管内皮生长因子基因转染能进一步增强大鼠梗死缺血区血管再生、改善室壁厚度和心功能. 揹景:骨髓間質榦細胞和血管內皮細胞生長因子移植具有促進血管再生、改善心功能的作用,但是二者聯閤應用是否優于單獨應用尚不清楚.目的:觀察同種異體骨髓間質榦細胞移植聯閤血管內皮生長因子基因轉染對大鼠急性心肌梗死血管再生和心功能的影響.設計:大鼠骨髓間質榦細胞培養採用單一樣本觀察;細胞移植和基因轉染採用隨機對照動物實驗.單位:華中科技大學協和醫院心內科,同濟醫學院心血管病研究所.材料:健康雄性Wistar大鼠94隻.錶達載體PAdTrack/VEGF165.方法:實驗于2004-06/2005-06在同濟醫學院心血管病研究所實驗室完成.①體外分離、純化、培養大鼠骨髓間質榦細胞,以5-溴-2'-脫氧尿苷標記細胞.②製備、抽提、純化、鑒定質粒PAdTrack/VEGF165.③結扎冠狀動脈建立急性心肌梗死模型2週後,隨機將其分為4組,每組均為12隻,榦細胞+質粒組、榦細胞組、質粒組、對照組,分彆進行心肌內大鼠骨髓間質榦細胞移植和/或VEGF165轉染、DMEM註射.④4週後行免疫組織化學和超聲心動圖檢查.主要觀察指標:①各組大鼠梗死及缺血區免疫組織化學和囌木精-伊紅染色檢查.②血管計數.③超聲心動圖檢查.結果:48隻大鼠進入結果分析.①榦細胞+質粒組和榦細胞組梗死及缺血心肌處可見大量5-溴-2'-脫氧尿苷標記的移植細胞,其中缺血心肌處部分移植細胞分化為血管內皮細胞併形成新生毛細血管.②Ⅷ因子染色暘性的新生血管密度分佈為榦細胞+質粒組＞質粒組＞榦細胞組＞對照組(P均＜0.01).③細胞移植和基因轉染治療後室壁厚度和室壁運動幅度改善,射血分數值增加幅度為榦細胞+質粒組＞榦細胞組＞質粒組＞對照組(P均＜0.01).結論:同種異體骨髓間質榦細胞移植聯閤血管內皮生長因子基因轉染能進一步增彊大鼠梗死缺血區血管再生、改善室壁厚度和心功能.
배경:골수간질간세포화혈관내피세포생장인자이식구유촉진혈관재생、개선심공능적작용,단시이자연합응용시부우우단독응용상불청초.목적:관찰동충이체골수간질간세포이식연합혈관내피생장인자기인전염대대서급성심기경사혈관재생화심공능적영향.설계:대서골수간질간세포배양채용단일양본관찰;세포이식화기인전염채용수궤대조동물실험.단위:화중과기대학협화의원심내과,동제의학원심혈관병연구소.재료:건강웅성Wistar대서94지.표체재체PAdTrack/VEGF165.방법:실험우2004-06/2005-06재동제의학원심혈관병연구소실험실완성.①체외분리、순화、배양대서골수간질간세포,이5-추-2'-탈양뇨감표기세포.②제비、추제、순화、감정질립PAdTrack/VEGF165.③결찰관상동맥건립급성심기경사모형2주후,수궤장기분위4조,매조균위12지,간세포+질립조、간세포조、질립조、대조조,분별진행심기내대서골수간질간세포이식화/혹VEGF165전염、DMEM주사.④4주후행면역조직화학화초성심동도검사.주요관찰지표:①각조대서경사급결혈구면역조직화학화소목정-이홍염색검사.②혈관계수.③초성심동도검사.결과:48지대서진입결과분석.①간세포+질립조화간세포조경사급결혈심기처가견대량5-추-2'-탈양뇨감표기적이식세포,기중결혈심기처부분이식세포분화위혈관내피세포병형성신생모세혈관.②Ⅷ인자염색양성적신생혈관밀도분포위간세포+질립조＞질립조＞간세포조＞대조조(P균＜0.01).③세포이식화기인전염치료후실벽후도화실벽운동폭도개선,사혈분수치증가폭도위간세포+질립조＞간세포조＞질립조＞대조조(P균＜0.01).결론:동충이체골수간질간세포이식연합혈관내피생장인자기인전염능진일보증강대서경사결혈구혈관재생、개선실벽후도화심공능.
BACKGROUND: Transplantation of bone marrow mesenchymal stem cell and vascular endothelial growth factor(VEGF) can promote vascular regeneration and improve heart function. However, whether the combined application is superior to single application or not is still unclear.OBJECTIVE: To observe the effect of allogenetic bone manow stem cells transplantation combined with VEGF transfection on vascular regeneration and heart function of rats with acute myocardial infarction.DESIGN: Simple sample observation was used in culturing bone marrow mesenchymal stem cell of rats; Randomized controlled animal experiment was used in cell transplantation and gene transfection.SETTING: Department of Cardiology, Union Hospital of Huazhong University of Science and Technology; Cardiovascular Institute of Tongji Medical College MATERIALS: Totally 94 healthy male Wistar rats and expression vector PAdTrack/VEGF165 were used in this experiment.METHODS: This experiment was carried out at Cardiovascular Institute of Tongji Medical College between June 2004 and June 2005. ①Bone marrow mesenchymal stem cells of rats were isolated, purified and cultured in vitro, then labeled with bromodeoxyuridine(BrdU). ② Preparation , extraction, purification and identification of plasmid PAdTrack/VEGF165. ③Two weeks after coronary artery was ligated to create acute myocardial infarction model, rats were randomly divided into 4 groups (n=12 in each group): stem cell + plasmid group(50 μL BrdU-labeled bone marrow mesenchymal stem cell solution and 100 μL plasmid PAdTrack/VEGF165 were injected into the rats through multiple sites), stem cell group (50 μL bone marrow mesenchymal stem cell solution was injected through multiple sites), plasmid group (100 μL plasmid PAdTrack/VEGF165 was injected through multiple sites) , control group(100 μL serum-free DMEM was injected through multiple sites). ④ Immunohistochemistry andechocardiography were performed 4 weeks later.MAIN OUTCOME MEASURES: ①Immunohistochemical and haematoxylin-eosin stainings were conducted in the infarcted and ischemic areas of rats in each group; ② Blood vessel counts; ③Echocardiography.RESULTS: Totally 48 rats entered the stage of result analysis. ① BrdUlabeled transplanted cells could be seen at the infarcted and ischemic myocardium in the stem cell+plasmid group and stem cell group. Some transplanted cells at ischemic myocardium differentiated into vascular endothelial cells and formed newborn blood capillary. ②Density of Ⅷ factor positively-stained newborn blood capillary took stem cell +plasmid group ＞ plasmid group ＞ stem cell group ＞ control group in order (all P＜ 0.01).③Wall thickness and wall motion range improved after cell transplantation and gene transfection therapy. The increased range of ejection fraction took stem cell +plasmid group ＞ stem cell group ＞ plasmid group ＞ control group in order (all P ＜ 0.01) .CONCLUSION: Allogenic bone marrow mesenchymal stem cell transplantation and VEGF gene transfection could further boost vascular regeneration of infarcted ischemic area and improve wall thickness and heart function of rats.