大鼠骨髓间充质干细胞移植治疗实验性大鼠股动脉闭塞症的研究  

Treatment of experimental femoral artery obliteransin rat by transplantation with Bone Marrow Mesenchymal Stem Cells from bone marrow

作　　者：张英杰[1] 宋振岚[2] 李玉云[1] 陈乐[1] 郝艳梅[1] 

机构地区：[1]蚌埠医学院检验系临检教研室,安徽蚌埠233030 [2]大连医科大学附属第二医院血液内科,辽宁大连116023

出　　处：《中国医疗前沿（学术版）》2008年第6期22-24,127,共4页China Healthcare Innovation

摘　　要：目的探讨骨髓间充质干细胞(MSCs)单独或与必要的诱导因子组合移植治疗实验性大鼠股动脉闭塞症,为临床应用间充质干细胞移植治疗糖尿病足等血管闭塞性疾病提供实验依据。方法密度梯度离心结合贴壁培养法分离纯化MSCs,结扎并断离大鼠双侧股动脉,复制成大鼠股动脉闭塞症模型。分别在大鼠后肢局部缺血肌肉通过多点注射的方法注射生理盐水(A组),BrdU标记的MSCs悬液(B组);BrdU标记的MSCs与利于血管新生的细胞因子悬液(C组)。14天处死大鼠,取双下肢缺血部位肌肉做HE染色、免疫组织化学(FVIII、BrdU)检测。结果密度梯度离心结合贴壁培养法能有效分离纯化MSCs,移植后14d免疫组化染色发现,肌束间B组、C组有BrdU阳性的血管,而A组无BrdU阳性的血管。平均每视野FVIII染色阳性的血管数目:A组为2.8±0.6个/HP,B组为8.4±2.3个/HP,C组为12.6±2.9个/HP;B组明显高于A组(p<0.01),C组明显高于B组(p<0.05)。结论密度梯度离心结合贴壁培养法能有效分离纯化MSCs;在体外扩增的MSCs移植于缺血的活体组织可以发育成血管,从而改善缺血部位的血液循环。无论是MSCs单独移植还是与必要的细胞因子组合移植其血管再生作用都超过大鼠自身在缺血环境中的血管重建或再生。但MSCs与必要的细胞因子组合的血管再生作用明显超过MSCs单独移植。Objective To explore transplantation with rat Marrow Mesenchymal Stem Cells(MSCs) only or MSCs with necessary inductor treat experimental femoral artery obliterans in rat. Methods MSCs were isolated from healthy rat and purifided by using density gradient centrifugation and adherent culture methods. Experimental rats models with both femoral arteries obliterans were successful made by ligating and excising both femoral arteries as well as their branches. A fluid (normal saline) as control were injected into ischemia hindlimbs of rats by intramusculer injection as group A, the same volume of B fluid (MSCs labled with BrdU ) were similarly injected into group B, C fluid(MSCs labled with BrdU, VEGF and bFGF ) were injected into group C. 14 days later, Hindlimb muscle was observed in sections stained with hematoxylin and eosin, FVIII and BrdU was investigated using immunohistochemical methods. Results MSCs can be isolated and purified by density gradient centrifugation and adherent culture methods. After 14 days, blood vessels with positive staining of BrdU in hindlimb were found in group B and C but not in A. The numbers of blood vessels with positive staining of Factor VIII in hindlimb at 14d after transplantation in group C(12.6±2.9) were more than that in group B(8.4±2.3) (p<0.05), and group B obviously more than that in group A(2.8±0.6) (p<0.01).

关 键 词：骨髓间充质干细胞 血管内皮细胞 股动脉闭塞症 血管再生 

分 类 号：R543.5[医药卫生—心血管疾病]