| 小肠RNA对缺血再灌注条件下移植小型猪骨髓内皮祖细胞的体外干预效应观察 |
| 投稿时间：2010-08-23 点此下载全文 |
| 引用本文：程康,王海昌,周祥,宗小娟,曾桂英.小肠RNA对缺血再灌注条件下移植小型猪骨髓内皮祖细胞的体外干预效应观察[J].医学研究杂志,2010,39(12):29-34 |
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| 基金项目:国家自然科学基金资助项目（30370581） |
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| 中文摘要:目的观察小肠RNA对小型猪骨髓内皮祖细胞（EPCs）生长的影响及适宜浓度，以及小肠RNA对缺血再灌注条件下移植EPCs的干预效应。方法通过Ficoll方法分离，差速贴壁纯化，诱导生成猪EPCs，分离提纯大鼠小肠RNA。设立不同浓度RNA组，观察其对EPCs生长增生等细胞功能的影响；依据适宜浓度，设立小肠RNA预处理组、缺血再灌注后加小肠RNA组、单纯小肠RNA孵育组、单纯缺血再灌注组和正常对照组，观察细胞生长曲线和LDH变化，还原酶法测量NO、NOS，观察EPCs的分泌功能。应用流式细胞仪观察EPCs凋亡情况，Western Blotting观察EPCs表面Flk-1的表达差异。结果小肠RNA在20μg/ml左右浓度促EPCs增生作用最明显；小肠RNA预处理组EPCs的Flk-1水平上调，EPCs凋亡较少，LDH和iNOS生成明显低于单纯缺血再灌注组和缺血再灌注后添加小肠RNA组，但高于正常对照组和单纯小肠RNA处理组。结论适宜浓度的小肠RNA可以促进EPCs的增生；小肠RNA预处理可以通过上调EPCs表面Flk-1水平，减少凋亡和分泌不利因子等机制增强EPCs抵抗缺血再灌注后移植的微环境改变的不利影响。 |
| 中文关键词:内皮祖细胞 缺血再灌注损伤 小肠RNA |
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| Interfering Effect of Intestine RNA on Marrow-derived Endothelial Progenitor Cells of the Minipig Endured Ischemia and Reperfusion |
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| Abstract:ObjectiveTo observe the growth and ideal content of marrow-derived endothelial progenitor cells (EPCs) of the minipig affected by intestine RNA, and study the interfering effect of intestine RNA on EPCs transplantation endured ischemia reperfusion injury (IRI) .MethodsWe induced minipig EPCs by Ficoll separation method and purification based on different adherence rate, and separated the purified rat intestine RNA. We set different content of RNA groups to observe the effect on EPCs growth:group of intestine RNA pretreatment， group of adding intestine RNA after ischemia and reperfusion,group of incubation only with intestine RNA,group of simply ischemia and reperfusion,and normal control.The growth curve of the cells and content of LDH were detected. We measured NO, NOS with the deoxidize-enzyme methods to observe the secretion function of EPCs. Apoptosis situation was observed with flow cytometry, and Flk-1 expression on the surface of EPCs with western blotting.ResultsThe obviously stimulative effect of intestine RNA on EPCs growth was about 20μg/ml. The expression of Flk-1 was up-regulated in the group pretreated with intestine RNA, with less apoptosis of EPCs. The amount of LDH and iNOS formation was significantly less than that in the simply ischemia and reperfusion group and treatment with adding intestine RNA after ischemia and reperfusion group, but higher than that in the normal control group and simply intestine RNA treatment group. ConclusionIdeal content of intestine RNA can promote the proliferation of EPCs. Intestine RNA pretreatment may protect EPCs by increasing the tolerance of EPCs to IRI and change of microenvironment by up-regulation of Flk-1, reducing apoptosis and down-regulation of bad factor formation. |
| keywords:Endothelial progenitor cells Ischemia-reperfusion injury Intestine RNA |
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