TNF-α对大鼠骨髓间充质干细胞黏附分子L-选择素表达的影响

目的 研究大鼠骨髓间充质干细胞（mesenchymal stem cell,MSCs）经不同浓度的肿瘤坏死因子-α（tumor necrosis factor,TNF-α）预处理后,表面的黏附分子L-选择素（L-selectin,CD62L）的表达变化以及其表达变化与细胞因子浓度之间的关系.方法 采用等密度梯度离心法收集大鼠MSCs并用贴壁培养法进行扩增纯化,经免疫组化法鉴定后,加入不同浓度TNF-α分别作用24小时,将其消化分离和标记后,采用流式细胞仪（flow cytometer,FCM）对其表面的CD62L的表达进行检测.结果 经TNF-α预处理24小时的大鼠MSCs经流式细胞仪检测,其表面CD62L成阳性表达的细胞占所检测细胞的百分比与未加入任何细胞因子预处理的空白组大鼠MSC相比较,差异显著,不同浓度组间比较,差异显著.结论 经不同浓度TNF-α预处理的大鼠MSCs表面CD62L表达较空白组要高,并且随着TNF-α浓度的升高大鼠MSCs表面CD62L的表达也相应升高.

The expression of L - selectin on rat mesenchymal stem cells after pretreated with TNF -α

Objective To study the expression change of L- selectin （CD62L） on rat mesenchymal stem cells （MSCs） after pretreated them with different concentration of tumor necrosis factor - α（ TNF - α） and also the relationship be- tween the expression change and the cytokine concentration. Methods Rat MSCs were isolated by density gradient centrlfuga- tion and were amplified and purified by adherent method. Ceils was identified by indirect immunofluorescence. The different concentration of TNF - α were added in different apertures and reacted 24 hours respectively. The rat MSCs were digested,sep- arated and labelled. After that, The expression of adhesion molecule CD62L on the surface of rat MSCs were detected by flow cytometer（FCM）. Results The rat MSCs which had pretreated with TNF -α, it＇s found that the percentage which CD62L was positive expressed in all cells compared with blank group which hadn＇t added any cytokine, the difference is significant; when compared among groups of different cytokine concentration. They were all statistically significant. Conclusion The ex- pression of CD62L on rat MSCs which has pretreated with different concentration of TNF - otis higher than blank group, and the expression increases with rising concentration.