Preliminary study of effect of erythropoietin pretreatment on enhancing directional homing ability of bone marrow mesenchymal stem cells in rats
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摘要:
目的 探讨促红细胞生成素（EPO）对大鼠骨髓间充质干细胞（BMSC）增殖及迁移能力的影响。 方法 将第5代BMSC分为5组，分别为对照组（不加EPO）及10、100、500、1 000 IU/mL EPO组，培养24 h和48 h后检测各组BMSC的增殖率、迁移能力以及趋化因子受体（CXCR）4的表达情况。将第5代BMSC分为BMSC组和EPO-BMSC组，培养48 h后观察EPO对两组BMSC的表面标志物、定向分化和细胞骨架形态的影响。 结果 EPO与BMSC共培养48 h后，与对照组比较，100、500 IU/mL EPO组BMSC增殖率和迁移能力增强，CXCR4蛋白表达量增高（均为P < 0.05）。EPO-BMSC组BMSC表面标志物的表达和定向分化能力未受EPO影响。在EPO-BMSC组中，大多数细胞的纤维骨架沿细胞的长轴排列，呈平行状。 结论 EPO可提高BMSC的增殖率、迁移能力和组织修复治疗能力，可能通过增加CXCR4的表达，促进其向损伤器官组织的定向归巢。 Abstract:Objective To evaluate the effect of erythropoietin (EPO) on the proliferation and migration of bone marrow mesenchymal stem cell (BMSC) in rats. Methods The 5th generation BMSCs were divided into the control (without EPO) and 10, 100, 500, 1 000 IU/mL EPO groups. After 24 h and 48 h of culture, the proliferation rate, migration ability and the expression levels of CXCR4 of BMSCs were detected in each group. The 5th generation BMSCs were further divided into BMSC and EPO-BMSC groups. After 48 h of culture, the effect of EPO upon surface markers, directional differentiation and cytoskeleton morphology of BMSCs were evaluated in both groups. Results After theco-culture of EPO and BMSCs for 48 h, the proliferation rate and migration ability of BMSCs were significantly enhanced, and the expression level of CXCR4 protein was significantly up-regulated in the 100 IU/mL and 500 IU/mL EPO groups compared with those in the control group (all P < 0.05). However, EPO exerted no effect upon the expression levels of surface markers and directional differentiation ability of BMSCs in the EPO-BMSC group. In the EPO-BMSC group, the fibrous skeleton of most BMSCs was arranged along the long axis in parallel. Conclusions EPO can improve the proliferation rate, migration ability and tissue repair capability of BMSCs, probably by promoting the directional homing of BMSCs to injured organs and tissues via up-regulating the expression level of CXCR4. -
Key words:
- Bone marrow mesenchymal stem cell /
- Transplantation /
- Erythropoietin /
- Homing /
- Migration /
- Proliferation /
- Chemokine receptor /
- Differentiation
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图 1 EPO对BMSC增殖率的影响
注：与对照组比较，aP < 0.05；与100 IU/mL EPO组比较，bP < 0.05。
Figure 1. Effects of EPO on proliferation rate of BMSC
图 2 EPO对BMSC迁移能力的影响（×100）
注：A图为空白对照，B图为对照组；C~F图分别为10、100、500、1 000 IU/mL EPO组。
Figure 2. Effects of EPO on migration of BMSC
图 3 EPO对BMSC中CXCR4表达水平的影响
注：与对照组比较，aP < 0.05；与100 IU/mL EPO组比较，bP < 0.05。
Figure 3. Effects of EPO on CXCR4 expression level in BMSC
图 4 EPO对BMSC表面标志物表达的影响
注：A~C图为BMSC组BMSC表面标志物表达情况；D~F图为EPO-BMSC组BMSC表面标志物表达情况。
Figure 4. Effects of EPO on expression of surface markers of BMSC
图 5 EPO对BMSC定向分化的影响
注：A、B图分别为BMSC组和EPO-BMSC组的成骨诱导结果（硝酸银，×100）；C、D图分别为BMSC组和EPO-BMSC组的成脂诱导结果（油红O，×100）。
Figure 5. Effects of EPO on directional differentiation of BMSC
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