骨髓基质细胞迁移机制的体外实验研究

发布时间：2018-02-28 09:51

本文关键词： 骨髓基质细胞趋化因子迁移　出处：《青岛大学》2009年硕士论文　论文类型：学位论文

【摘要】： 目的:观察体外条件下趋化因子CXCL-8、CXCL-16对大鼠骨髓基质细胞迁移的趋化作用。方法:采用全骨髓法培养成年Wistar大鼠骨髓基质细胞,取第5代骨髓基质细胞行流式细胞仪鉴定;然后通过细胞免疫荧光及RT-PCR的方法检测骨髓基质细胞表达趋化因子受体CXCR1、CXCR6的情况,利用Transwell小室法探讨趋化因子CXCL-8、CXCL-16对骨髓基质细胞的体外趋化作用及其特异性。结果:第5代骨髓基质细胞的表面抗原CD29的阳性率为99.18%,CD45的阳性率为4.35%;骨髓基质细胞趋化因子受体CXCR1和CXCR6呈阳性表达,位于细胞膜和细胞浆中,RT-PCR琼脂糖凝胶电泳后分别出现215 bp、342bp特异性扩增条带。与对照组比较,加入5～500 ng/ml趋化因子CXCL-8或CXCL-16后,骨髓基质细胞迁移数均明显增加(P0.05)。加入抗CXCR1或CXCR6多克隆抗体后,骨髓基质细胞迁移数较250 ng/ml趋化因子CXCL-8和CXCL-16组明显减少(P0.05)。结论:体外条件下趋化因子CXCL-8、CXCL-16在5～500 ng/ml质量浓度范围可趋化骨髓基质细胞迁移,封闭趋化因子受体CXCR1或CXCR6后其趋化迁移作用明显减弱。CXCL-8＼CXCR1和CXCL-16＼CXCR6通路参与骨髓基质细胞体外迁移,为进一步研究骨髓基质细胞的迁移机制提供了理论依据。
[Abstract]:Aim: to observe the chemotactic effect of CXCL-8 CXCL-16 on the migration of rat bone marrow stromal cells in vitro. Methods: bone marrow stromal cells (BMSCs) of adult Wistar rats were cultured by whole bone marrow method, and bone marrow stromal cells of the 5th passage were identified by flow cytometry. Then the expression of chemokine receptor CXCR1 and CXCR6 in bone marrow stromal cells was detected by cellular immunofluorescence and RT-PCR. The chemotactic effect and specificity of CXCL-8 and CXCL-16 on bone marrow stromal cells were investigated by Transwell chamber method. Results: the positive rate of surface antigen CD29 of bone marrow stromal cells was 99.18% and the positive rate of CD45 was 4.35. The expression of chemokine receptor CXCR1 and CXCR6 in bone marrow stromal cells was positive. After RT-PCR agarose gel electrophoresis, a specific amplified band of 215 BP was found in the cell membrane and cytoplasm. Compared with the control group, the 5 ng/ml chemokine CXCL-8 or CXCL-16 was added. The migration number of bone marrow stromal cells increased significantly (P 0.05), and the number of bone marrow stromal cells migration decreased significantly compared with that of 250 ng/ml chemokine CXCL-8 and CXCL-16 groups after the addition of anti CXCR1 or CXCR6 polyclonal antibodies. Conclusion: the chemokine CXCL-8 CXCL-16 can induce the migration of bone marrow stromal cells in the concentration range of 5500 ng/ml in vitro. The chemokine migration of bone marrow stromal cells after blocking chemokine receptor CXCR1 or CXCR6 is obviously weakened. CXCL-8\ CXCR1 and CXCL-16\ CXCR6 pathway are involved in the migration of bone marrow stromal cells in vitro. It provides a theoretical basis for further studying the migration mechanism of bone marrow stromal cells.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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