CAJ | 학술논문

背景:常规药物治疗、介入或血管旁路移植手术对下肢动脉闭塞症的远期疗效均不理想.近年来采用干细胞的血管再生疗法在梗死部位修复或重新建立有效侧支循环逐渐成为可能.目的:验证骨髓间充质干细胞自体移植对兔缺血后肢血管再生的促进作用.设计、时间及地点:随机对照动物实验,于2005 08/2006-11在江苏省血吸寄生虫病防治研究所完成.材料:选用新西兰大白兔8只,制备兔后肢缺血模犁.按随机数字表法分为2组,实验组及对照组各4只.方法:分离培养实验组兔骨髓间充质干细胞,5-溴-2-脱氧尿苷标记,将骨髓间克质干细胞制成悬液注射于实验组兔后肢缺血部位;对照组注射等量生理盐水.主要观察指标:2周后行兔股动脉二维及彩色多普勒超声检测,观察移植前后的血管内径、血流峰值速度及血流加速时间等;取缺血部位肌肉组织,通过免疫荧光及苏木精一伊红染色观察移植细胞分布及血管生成情况.结果:细胞移植2周后实验组兔的股动脉内径、血流峰值速度均大于对照组(P<0.01),血流加速时间短于对照组(P<0.01).免疫荧光染色结果显示实验组兔移植部位有抗5-溴-2-脱氧尿苷染色阳性细胞存在:苏木精-伊红染色结果显示实验组缺血部位血管密度高于对照组.结论:骨髓间充质干细胞具有促进血管生成的作用,自体移植可望成为一种简单的、有效的治疗下肢缺血的方法.
배경:상규약물치료、개입혹혈관방로이식수술대하지동맥폐새증적원기료효균불이상.근년래채용간세포적혈관재생요법재경사부위수복혹중신건립유효측지순배축점성위가능.목적:험증골수간충질간세포자체이식대토결혈후지혈관재생적촉진작용.설계、시간급지점:수궤대조동물실험,우2005 08/2006-11재강소성혈흡기생충병방치연구소완성.재료:선용신서란대백토8지,제비토후지결혈모리.안수궤수자표법분위2조,실험조급대조조각4지.방법:분리배양실험조토골수간충질간세포,5-추-2-탈양뇨감표기,장골수간극질간세포제성현액주사우실험조토후지결혈부위;대조조주사등량생리염수.주요관찰지표:2주후행토고동맥이유급채색다보륵초성검측,관찰이식전후적혈관내경、혈류봉치속도급혈류가속시간등;취결혈부위기육조직,통과면역형광급소목정일이홍염색관찰이식세포분포급혈관생성정황.결과:세포이식2주후실험조토적고동맥내경、혈류봉치속도균대우대조조(P<0.01),혈류가속시간단우대조조(P<0.01).면역형광염색결과현시실험조토이식부위유항5-추-2-탈양뇨감염색양성세포존재:소목정-이홍염색결과현시실험조결혈부위혈관밀도고우대조조.결론:골수간충질간세포구유촉진혈관생성적작용,자체이식가망성위일충간단적、유효적치료하지결혈적방법.
BACKGROUND: Longterm therapeutic effects of routine drug treatment, intervention or vascular bypass transplantation on lower extremity arterial occlusion are not ideal. During recent years, angiogenesis of stem cells possibly becones a new method to repair or rebuild an effective collateral circulation at infarct regions.OBJECTIVE: To verify the effect of autologous bone marrow mesenchymal stem cell transplantation on promoting angiogenesis in rabbit ischemic hind limbs.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed in Jiangsu Institute of Schistosomiasis Control between August 2005 and November 2006. MATERIALS: Eight rabbits were used to prepare ischemic models of hind limbs, and then they were randomly divided into experimental group (n=4) and control group (n=4).METHODS: Bone marrow mesenchymal stem cells were isolated and cultured from New Zealand rabbits in the experimental group, and they were then marked 5-bromo-2-deoxyuridine (Brdu). Suspension of bone marrow mesenchymal stem cells was injected into the ischemic hind limbs in the experimental group, while the same volume of saline was injected into the controls. MAIN OUTCOME MEASURES: After two weeks, two-dimensional and color Doppler ultrasound detection was used on rabbit femoral artery to measure inner diameter of blood vessel, peak velocity and acceleration time of blood flow before and after transplantation. Muscle tissues were obtained from ischemic regions to observe distribution of transplant cells and state of angiogenesis using immunofluorescence staining and hematoxylin-eosin (HE) staining. RESULTS: Two weeks after transplantation, the inner diameter of femoral artery and the peak velocity of blood flow in the experimental group were higher than those in the control group (P < 0.01), but the acceleration time of blood flow was shorter than that in the control group (P < 0.01). Lmmunofluorescence staining showed that anti-Brdu-staining positive cells were found out in transplant part in the experimental group; while HE staining indicated that vessel density of ischemic region in the experimental group was higher than that in the control group. CONCLUSION: Bone marrow mesenchymal stem cells can promote angiogenesis, while autologous bone marrow mesenchymai stem cell transplantation will become a simple and effective method to treat lower limb ischemia.