Ren Ling, Zhou Lin, Xue Li, Fan Zhongcai
Department of Cardiology, First People's Hospital of Yibin, Yibin 644000, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2009 Oct;26(5):1077-81.
To investigate effects of tumor necrosis factor-alpha(TNF-alpha) and granulocyte-colony stimulation factor(G-CSF) on the transplantation rate of rabbit mesenchymal stem cells (MSCs) through intracoronary injection. Japanese white ears rabbits (2-3 months old) were used as myocardial injury models by intravenously injecting adriamycin, then they were divided into three groups randomly: (1) Blank group, (2) TNF-alpha group; (3) G-CSF group. There were 7 rabbits each group. Before transplantation, MSCs and rabbit models were neither intervened by cytokine in blank group; in TNF-alpha group, rabbits were injected TNF-alpha (5 microg/kg) through ear-edge vein one day before transplantation, MSCs were pretreated by TNF-alpha (10ng/ml)12 hours before transplantation. In G-CSF group, rabbits were injected G-CSF(10 microg/kg) intramuscular each day 3 days before transplantation. MSCs were pretreated by G-CSF(10 ng/ ml) 12 hours before transplantation. Separated by density gradient centrifugation and labeled by 4,6-biamidine-2-phenyliodole (DAPI) after cells expanded in vitro, MSCs were then transplanted through the root of aorta when the ascending aorta occluded above the sinus aortae. Myocardial pathological sections were made after 4 weeks and count transplanted cells under laser scanning confocal microscope and statiscally analyze. The amount of cells transplanted in rabbit myocardial were much more in TNF-alpha group G-CSF group than in blank group, there was statistically significant difference (P < 0.001). It sugested that TNF-alpha and G-CSF intervened the cultured MSCs, and animal model of myocardial injury could elevate the transplantation rate of intracoronary injection rabbit mesenchymal stem cells. The mechanism may have relation to that cytokine can enhance the attachment of MSCs to cardiac microvascular endothelium (CMVE).
探讨肿瘤坏死因子-α（TNF-α）和粒细胞集落刺激因子（G-CSF）对兔间充质干细胞（MSCs）经冠状动脉注射移植率的影响。选用2-3月龄日本大耳白兔，通过静脉注射阿霉素建立心肌损伤模型，然后将其随机分为三组：（1）空白组；（2）TNF-α组；（3）G-CSF组。每组7只兔。移植前，空白组的MSCs和兔模型均未用细胞因子干预；TNF-α组在移植前1天经耳缘静脉注射TNF-α（5μg/kg），MSCs在移植前12小时用TNF-α（10ng/ml）预处理。G-CSF组在移植前3天每天肌肉注射G-CSF（10μg/kg）。MSCs在移植前12小时用G-CSF（10ng/ml）预处理。体外培养扩增后的细胞经密度梯度离心分离，并用4,6-二脒基-2-苯基吲哚（DAPI）标记，然后在升主动脉窦上方阻断升主动脉时经主动脉根部移植MSCs。4周后制作心肌病理切片，在激光扫描共聚焦显微镜下计数移植细胞并进行统计学分析。TNF-α组和G-CSF组兔心肌内移植的细胞数量明显多于空白组，差异有统计学意义（P<0.001）。提示TNF-α和G-CSF对培养的MSCs有干预作用，心肌损伤动物模型可提高兔间充质干细胞经冠状动脉注射的移植率。其机制可能与细胞因子能增强MSCs与心肌微血管内皮（CMVE）的黏附有关。
