CAJ | 학술논문

目的：探讨外源骨桥蛋白（OPN）对小鼠骨髓间充质干细胞（MSCs）迁移能力的影响。方法采用OPN -/-小鼠和野生型C57小鼠，进行MSCs的原代分离培养，流式细胞术鉴定分拣MSCs细胞传代培养；Transwell迁移实验检测 OPN 是否能够诱导MSCs定向迁移；蛋白免疫印迹法（Western blot）检测OPN、CD44和Integrin β1相关蛋白的表达变化。结果传代后的细胞形态符合 MSCs特征，细胞表达 CD44和 CD105，但不表达 CD34，符合 MSCs表面标记抗原的一般规律。0．5μg/mL的OPN能增加体外MSCs的细胞迁移，而野生型C57小鼠MSCs细胞迁移为最多。同时，这一趋势与OPN 作用时间正相关，与OPN -/-小鼠相比，差异均有统计学意义（P＜0．05）。0．5μg/mL重组OPN蛋白量明显增加，但仍低于野生型C57小鼠，差异有统计学意义（P＜0．01）。若以OPN -/-小鼠为基数100，0．5μg/mL重组OPN组CD44、Integrinβ1蛋白表达增多，差异有统计学意义（P＜0．01）；而野生型C57小鼠细胞CD44、Integrinβ1蛋白表达最多，差异有统计学意义（P＜0．01）。结论 OPN 可以通过上调 CD44、Integrinβ1的表达，促进MSCs定向迁移。
목적：탐토외원골교단백（OPN）대소서골수간충질간세포（MSCs）천이능력적영향。방법채용OPN -/-소서화야생형C57소서，진행MSCs적원대분리배양，류식세포술감정분간MSCs세포전대배양；Transwell천이실험검측 OPN 시부능구유도MSCs정향천이；단백면역인적법（Western blot）검측OPN、CD44화Integrin β1상관단백적표체변화。결과전대후적세포형태부합 MSCs특정，세포표체 CD44화 CD105，단불표체 CD34，부합 MSCs표면표기항원적일반규률。0．5μg/mL적OPN능증가체외MSCs적세포천이，이야생형C57소서MSCs세포천이위최다。동시，저일추세여OPN 작용시간정상관，여OPN -/-소서상비，차이균유통계학의의（P＜0．05）。0．5μg/mL중조OPN단백량명현증가，단잉저우야생형C57소서，차이유통계학의의（P＜0．01）。약이OPN -/-소서위기수100，0．5μg/mL중조OPN조CD44、Integrinβ1단백표체증다，차이유통계학의의（P＜0．01）；이야생형C57소서세포CD44、Integrinβ1단백표체최다，차이유통계학의의（P＜0．01）。결론 OPN 가이통과상조 CD44、Integrinβ1적표체，촉진MSCs정향천이。
Objective To investigate the effects of exogenous osteopontin (OPN ) the migration of bone marrow mesenchymal stem cells(MSCs) in mice .Methods Using the OPN -/- and wild-type C57 mice ,the MSCs were isolated and cultured .These cells were analysed and sorted by flow cytometry .Then ,Transwell migration assay detected whether OPN can induce the migration of these MSCs .Finally ,the method of Western blot was used to detect the changes of expression of OPN ,CD44 and Integrinβ1 pro-teins .Results The morphology and features of MSCs were proved correctly on 3 passages ,which these cells expressed the proteins of CD44 and CD105 ,but not CD34 .The surface marker antigens were accorded with the general features of MSCs .Compared with OPN -/- mice ,the MSCs significantly increased cell migration in groups of 0 .5 μg/mL OPN and wild type C57 mice ,which was positively related to the times using OPN .Using recombinant OPN protein(0 .5μg/mL) ,expression of related proteins was signifi-cantly increased ,but still lower than the group of wild type mice (P<0 .01) .Compared with OPN -/- mice ,expression of OPN , CD44 ,and Integrinβ1 protein increased significantly different (P<0 .01) in groups of 0 .5 μg/mL OPN and wild type C57 mice . Conclusion OPN can up-regulate the expression of CD44 and Integrinβ1 proteins and promote MSCs migration .