發(fā)布時間：2018-04-27 10:50

  本文選題：臍帶間充質干細胞 + 低劑量X射線�。� 參考：《蘭州大學》2011年碩士論文

【摘要】：目的：研究健康人臍帶間充質干細胞(human umbilical cord mesenchymal stem cells, hucMSCs)中趨化因子-1受體CXCR4的表達特征；探討低劑量X射線照射(LDR)對hucMSCs增殖、成脂成骨分化及基質衍生因子1受體CXCR4表達的影響。 方法：采用組織塊貼壁法從健康人臍帶中培養(yǎng)出成纖維樣細胞；流式細胞術檢測hucMSCs表面CD44、CD29、CD34、CD45、HLA-DR的表達；分別采用流式細胞儀及實時定量PCR檢測P2-P5代hucMSCs中CXCR4受體及mRNA的表達；MTT法檢測LDR對hucMSCs增殖的影響；觀察LDR對hucMSCs成脂成骨分化的影響；通過實時定量PCR及流式細胞儀分別檢測LDR后基質衍生因子1受體CXCR4mRNA及蛋白的量-效和時-效關系。 結果：流式細胞術結果顯示：hucMSCs高表達CD44、CD29,而CD34、CD45、HLA-DR呈陰性表達；P2-P5代hucMSCs中均有CXCR4蛋白表達,分別為(89.82±0.62)%、(86.87±1.32)%、(80.50±4.46)%、(70.10±0.68)%。P2-P5代hucMSCs中CXCR4mRNA均有表達,且P3-P5代CXCR4mRNA表達量分別為P2代的0.5585±0.0875倍、0.6205±0.1377倍、0.4634±0.0447倍。低劑量照射后,不同照射劑量組在各個時間段細胞增殖均高于對照組(P0.05),其中以100mGy組促進細胞增殖作用最明顯。照射組成脂誘導48 h后細胞內即有脂滴出現,且第21天成脂率明顯高于對照組(P0.05)。成骨誘導后,照射組堿性磷酸酶(ALP)活性明顯高于對照組(P0.05),且成骨細胞核結合因子α1mRNA水平明顯增高(P0.05)。實時定量PCR及流式細胞儀結果顯示：LDR促進了hucMSCs中CXCR4的表達,且這種表達呈現出一定的照射劑量及時間依賴性。 結論：健康人臍帶中培養(yǎng)的P2-P5代hucMSCs均表達CXCR4,且其表達量隨傳代數增加而呈降低趨勢；與P2代相比,P3-P5代hucMSCs CXCR4 mRNA表達均有所降低,但P3-P5各代間表達相對穩(wěn)定；LDR可以促進hucMSCs增殖,并加速其向成脂、成骨細胞分化；LDR可誘導hucMSCs中CXCR4的表達。
[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. 緇撴灉:嫻佸紡緇嗚優(yōu)鏈粨鏋滄樉紺，

本文編號：1810433