發(fā)布時(shí)間：2018-04-03 02:07

  本文選題：骨髓間質(zhì)干細(xì)胞　切入點(diǎn)：膠質(zhì)細(xì)胞　出處：《福建醫(yī)科大學(xué)》2005年碩士論文

【摘要】：目的研究趨化因子SDF-1α在體內(nèi)外對(duì)原代培養(yǎng)的骨髓間質(zhì)干細(xì)胞、星形膠質(zhì)細(xì)胞和N9 小膠質(zhì)細(xì)胞系以及分離的中性粒細(xì)胞的趨化作用,從而探討腦損傷和細(xì)胞移植時(shí)SDF-1α在細(xì)胞遷移中的可能機(jī)制。 方法體外實(shí)驗(yàn)利用SDF-1α作為趨化因子加入到48孔微遷移板下室,把上述四種細(xì)胞加入到上室,抗體阻斷實(shí)驗(yàn)把這四種細(xì)胞與抗CXCR4 多抗共孵育后加入到上室,觀察細(xì)胞遷移,計(jì)算細(xì)胞的遷移指數(shù)。同時(shí)用RT-PCR 方法觀察N9 小膠質(zhì)細(xì)胞CXCR4 基因表達(dá)的情況。體內(nèi)實(shí)驗(yàn)采用大鼠大腦中動(dòng)脈線栓模型,把骨髓間質(zhì)干細(xì)胞、中性粒細(xì)胞以及N9 小膠質(zhì)細(xì)胞與抗CXCR4 多抗共孵育后的這3 種細(xì)胞分別通過(guò)大鼠尾靜脈移植入體內(nèi),腦組織切片后熒光顯微鏡下觀察腦損傷部位植入細(xì)胞的趨化情況。 結(jié)果體外實(shí)驗(yàn)時(shí),SDF-1α作用后,骨髓間質(zhì)干細(xì)胞、N9 小膠質(zhì)細(xì)胞和中性粒細(xì)胞跨膜遷移的數(shù)目較對(duì)照組明顯增多,而星形膠質(zhì)細(xì)胞遷移的數(shù)目未見(jiàn)明顯變化�？笴XCR4 多抗孵育后骨髓間質(zhì)干細(xì)胞、N9 小膠質(zhì)細(xì)胞和中性粒細(xì)胞跨膜遷移的數(shù)目則明顯減少。RT-PCR 方法證實(shí)在正常情況下N9 小膠質(zhì)細(xì)胞有明顯的CXCR4 表達(dá)。體內(nèi)實(shí)驗(yàn)時(shí),有較多的骨髓間質(zhì)干細(xì)胞、N9 小膠質(zhì)細(xì)胞和中性粒細(xì)胞遷移到病灶部位,而空白對(duì)照組未見(jiàn)到遷移的細(xì)胞,陰性對(duì)照組僅見(jiàn)個(gè)別遷移的細(xì)胞。 結(jié)論趨化因子SDF-1α能通過(guò)受體CXCR4 發(fā)揮對(duì)骨髓間質(zhì)干細(xì)胞、N9 小膠質(zhì)細(xì)胞和中性粒細(xì)胞的趨化作用,對(duì)星形膠質(zhì)細(xì)胞沒(méi)有趨化作用。在體內(nèi),損傷的腦組織可能主要通過(guò)分泌SDF-1α增多作用于CXCR4 從而對(duì)炎癥細(xì)胞、小膠質(zhì)細(xì)胞或移植的骨髓間質(zhì)干細(xì)胞發(fā)揮趨化作用。
[Abstract]:Objective to investigate the chemotactic effects of chemokine SDF-1 偽 on primary cultured bone marrow mesenchymal stem cells, astrocytes, N9 microglial cells and isolated neutrophils in vitro and in vivo.To explore the possible mechanism of SDF-1 偽 in cell migration during brain injury and cell transplantation.Methods in vitro, SDF-1 偽 was used as chemokine in the 48 well micromigration chamber, and the above four cells were added to the upper chamber. The antibody blocking experiment incubated the four kinds of cells with anti CXCR4 polyclonal antibodies and then added them to the upper chamber to observe the migration of the cells.Cell migration index was calculated.At the same time, the expression of CXCR4 gene in N 9 microglial cells was observed by RT-PCR method.Bone marrow mesenchymal stem cells, neutrophilic granulocytes and N9 microglial cells were transplanted into the rat caudal vein after incubating with anti CXCR4 polyclonal antibodies in vivo.The chemotaxis of implanted cells in brain injury site was observed under fluorescence microscope after brain tissue section.Results the number of N9 microglia and neutrophil transmembrane migration of bone marrow mesenchymal stem cells was significantly increased after SDF-1 偽 treatment in vitro, but the number of astrocytes did not change.The number of transmembrane migration of N9 microglia and neutrophils in bone marrow mesenchymal stem cells (BMSCs) incubated with anti CXCR4 polyclonal antibodies was significantly decreased. RT-PCR showed that N9 microglia expressed CXCR4 in normal condition.In vivo, more bone marrow mesenchymal stem cells (BMSCs) N9 microglia and neutrophils migrated to the lesion site, but no migrating cells were found in the blank control group, while only individual migrating cells were found in the negative control group.Conclusion chemokine SDF-1 偽 can exert chemotaxis on N9 microglia and neutrophils in bone marrow mesenchymal stem cells through receptor CXCR4, but has no chemotactic effect on astrocytes.In vivo, the injured brain tissue may play a chemotaxis effect on inflammatory cells, microglia cells or bone marrow mesenchymal stem cells by secreting SDF-1 偽 increase in CXCR4.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R341

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本文編號(hào)：1703131