低剂量X射线照射对人脐带间充质干细胞生物学特性及基质衍生因子1受体CXCR4表达的影响

发布时间：2018-04-27 10:50

  本文选题：脐带间充质干细胞 + 低剂量X射线　； 参考：《兰州大学》2011年硕士论文

【摘要】：目的：研究健康人脐带间充质干细胞(human umbilical cord mesenchymal stem cells, hucMSCs)中趋化因子-1受体CXCR4的表达特征；探讨低剂量X射线照射(LDR)对hucMSCs增殖、成脂成骨分化及基质衍生因子1受体CXCR4表达的影响。 方法：采用组织块贴壁法从健康人脐带中培养出成纤维样细胞；流式细胞术检测hucMSCs表面CD44、CD29、CD34、CD45、HLA-DR的表达；分别采用流式细胞仪及实时定量PCR检测P2-P5代hucMSCs中CXCR4受体及mRNA的表达；MTT法检测LDR对hucMSCs增殖的影响；观察LDR对hucMSCs成脂成骨分化的影响；通过实时定量PCR及流式细胞仪分别检测LDR后基质衍生因子1受体CXCR4mRNA及蛋白的量-效和时-效关系。 结果：流式细胞术结果显示：hucMSCs高表达CD44、CD29,而CD34、CD45、HLA-DR呈阴性表达；P2-P5代hucMSCs中均有CXCR4蛋白表达,分别为(89.82±0.62)%、(86.87±1.32)%、(80.50±4.46)%、(70.10±0.68)%。P2-P5代hucMSCs中CXCR4mRNA均有表达,且P3-P5代CXCR4mRNA表达量分别为P2代的0.5585±0.0875倍、0.6205±0.1377倍、0.4634±0.0447倍。低剂量照射后,不同照射剂量组在各个时间段细胞增殖均高于对照组(P0.05),其中以100mGy组促进细胞增殖作用最明显。照射组成脂诱导48 h后细胞内即有脂滴出现,且第21天成脂率明显高于对照组(P0.05)。成骨诱导后,照射组碱性磷酸酶(ALP)活性明显高于对照组(P0.05),且成骨细胞核结合因子α1mRNA水平明显增高(P0.05)。实时定量PCR及流式细胞仪结果显示：LDR促进了hucMSCs中CXCR4的表达,且这种表达呈现出一定的照射剂量及时间依赖性。 结论：健康人脐带中培养的P2-P5代hucMSCs均表达CXCR4,且其表达量随传代数增加而呈降低趋势；与P2代相比,P3-P5代hucMSCs CXCR4 mRNA表达均有所降低,但P3-P5各代间表达相对稳定；LDR可以促进hucMSCs增殖,并加速其向成脂、成骨细胞分化；LDR可诱导hucMSCs中CXCR4的表达。
[Abstract]:Aim: to study the expression of chemokine 1 receptor (CXCR4) in human umbilical cord mesenchymal stem cells (umbilical cord mesenchymal stem cells, hucMSCs), and to investigate the effects of low dose X-ray irradiation on hucMSCs proliferation, adipogenic differentiation and CXCR4 expression of matrix derived factor 1 receptor. Methods: fibroblast-like cells were cultured from the umbilical cord of healthy subjects by tissue mass adherence, and the expression of HLA-DR on the surface of hucMSCs was detected by flow cytometry. Flow cytometry and real-time quantitative PCR were used to detect the expression of CXCR4 receptor and mRNA in hucMSCs of P2-P5 generation. The effect of LDR on hucMSCs proliferation and the effect of LDR on adipogenic differentiation of hucMSCs were observed. The dose-effect and time-response relationships of matrix derived factor-1 receptor (CXCR4mRNA) and protein after LDR were detected by real-time quantitative PCR and flow cytometry respectively. 缁撴灉:娴佸紡缁嗚優鏈�缁撴灉鏄剧ぃ�

本文编号：1810433