本文關(guān)鍵詞： 骨髓基質(zhì)細(xì)胞 趨化因子 遷移　出處：《青島大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:觀察體外條件下趨化因子CXCL-8、CXCL-16對(duì)大鼠骨髓基質(zhì)細(xì)胞遷移的趨化作用。 方法:采用全骨髓法培養(yǎng)成年Wistar大鼠骨髓基質(zhì)細(xì)胞,取第5代骨髓基質(zhì)細(xì)胞行流式細(xì)胞儀鑒定;然后通過細(xì)胞免疫熒光及RT-PCR的方法檢測(cè)骨髓基質(zhì)細(xì)胞表達(dá)趨化因子受體CXCR1、CXCR6的情況,利用Transwell小室法探討趨化因子CXCL-8、CXCL-16對(duì)骨髓基質(zhì)細(xì)胞的體外趨化作用及其特異性。 結(jié)果:第5代骨髓基質(zhì)細(xì)胞的表面抗原CD29的陽性率為99.18%,CD45的陽性率為4.35%;骨髓基質(zhì)細(xì)胞趨化因子受體CXCR1和CXCR6呈陽性表達(dá),位于細(xì)胞膜和細(xì)胞漿中,RT-PCR瓊脂糖凝膠電泳后分別出現(xiàn)215 bp、342bp特異性擴(kuò)增條帶。與對(duì)照組比較,加入5～500 ng/ml趨化因子CXCL-8或CXCL-16后,骨髓基質(zhì)細(xì)胞遷移數(shù)均明顯增加(P0.05)。加入抗CXCR1或CXCR6多克隆抗體后,骨髓基質(zhì)細(xì)胞遷移數(shù)較250 ng/ml趨化因子CXCL-8和CXCL-16組明顯減少(P0.05)。 結(jié)論:體外條件下趨化因子CXCL-8、CXCL-16在5～500 ng/ml質(zhì)量濃度范圍可趨化骨髓基質(zhì)細(xì)胞遷移,封閉趨化因子受體CXCR1或CXCR6后其趨化遷移作用明顯減弱。CXCL-8＼CXCR1和CXCL-16＼CXCR6通路參與骨髓基質(zhì)細(xì)胞體外遷移,為進(jìn)一步研究骨髓基質(zhì)細(xì)胞的遷移機(jī)制提供了理論依據(jù)。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

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本文編號(hào)：1546795