中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2012, Vol. 41 ›› Issue (2): 162–165.

• 临床医学 • 上一篇    下一篇

Toll样受体3和4在人骨髓间充质干细胞中的表达及其活化对细胞增殖的影响

何晓霞1, 2 , 白海1 , 杨国嵘3 , 马晓慧1 , 薛迪强2   

  1. 1.兰州军区总医院全军血液中心,兰州730050;2.甘肃省兰州市第二人民医院血液科,兰州730046;3.中国人民解放军第一医院病理科,兰州730030
  • 收稿日期:2012-09-20 修回日期:2012-09-20 出版日期:2012-02-20 发布日期:2012-09-21

Study about theExpression of Toll-like Receptor 3 (TLR3)and 4 TLR4 in Human Mesenchemal Mesenchymal Stem Cells Derived from Bone Marrow and about the Effects of tTheir Activation on Cell Proliferation

HE Xiao-xia1,2, BAI Hai1, YANG Guo-rong3, MA Xiao-Hui1, XUE Di-qiang2   

  1. 1 .Department of Hematology,General Hosipital of Lanzhou military Military Ddistrict of PLA, Lanzhou 730050 China; 2. Department of Hematology,The Second People’s Hospital of Lanzhou, Lanzhou 730046, China; 3 .Department of Pathology, The First Hospital of PLA, Lanzhou 730030, China
  • Received:2012-09-20 Revised:2012-09-20 Online:2012-02-20 Published:2012-09-21

摘要: 目的 研究Toll样受体(TLR)3和4在人骨髓间充质干细胞(BM-MSC)中的表达及其活化对细胞增殖的影响。方法 Ficoll法分离培养健康供体的BM-MSC;用流式细胞(FCM)检测BM-MSC中CD34、CD45、HLA-DR、CD44和CD71的表达;免疫细胞化学法检测爬片BM-MSC细胞中CD71的表达;RT-PCR法检测BM-MSC中TLR3和TLR4的表达;MTT法检测 POLY(IC)和LPS对BM-MSC增殖的影响。结果 FCM法发现体外分离培养的BM-MSC中CD34、CD45和HLA-DR表达呈阴性,CD44和CD71表达呈阳性;爬片BM-MSC细胞中CD71的表达呈阳性;BM-MSC中有较高水平的TLR 3和TLR4表达;体外培养第5天时,各浓度POLY(IC)组存活细胞数均少于血清组(P<0.01),而多于单纯培养基组(P <0.05),浓度和存活细胞数间存在半对数直线相关关系(r=?0.998,P<0.05);体外培养第7天时,中浓度和高浓度LPS组存活细胞数均少于血清组(P<0.05),而多于单纯培养基组(P<0.01)。结论 健康供体BM-MSC中存在高水平TLR3和TLR4 mRNA的表达;POLY(IC)和LPS通过TLR3和TLR4的介导影响BM-MSC的增殖。

关键词: 骨髓, 间充质干细胞, Toll样受体, 增殖, 骨髓, 间充质干细胞, Toll样受体, 增殖

Abstract: Objective To study the expression of Toll-like receptor 3 Toll-like receptor 3 (TLR3) and TLR44 in human mesenchymal mesenchemal stem cells derived from bone marrow(BM-MSC) and the effects of their activation on cell proliferation. Methods Mesenchymal stem cellsBM-MSC was were isolated from bone marrow of healthy donors by Ficoll method. Expression of CD34, CD45, HLA-DR, CD44 and CD71 in BM-MSC waswere shown by flow cytometry. CD71 was detected in BM-MSC by immunocytochemistry. Expression of TLR 3 and TLR4 4 was detected by the RT-PCR method. Effects of POLY(I:C) and LPS on proliferation of BM-MSC were studied by the MTT method. Results Expression of CD34, CD45 and HLA-DR in BM-MSC was negative while expression of CD44 and CD71 was positive by FCM. CD71 was localized in BM-MSC was positive by immunocytochemistry. High levels of TLR3 and TLR44 mRNA were found to be expressed in BM-MSC by RT-PCR. The vViable counts of different concentrations teams of POLY(I:C) were less than thatthose of 10% FBS team (P<0.01), but more than thatthose of DMEM team (P <0.05) after 5 days of after incubation while this type of semi- logarithmic relationship a semilogarithm linear correlation was found between concentration and viable count (r=?0.998, P<0.05). However, the viable counts of moderate and high concentrations teams of LPS were less than those that of 10% FBS team (P<0.05), but more than that those of DMEM team (P<0.05) after 7 days of after incubation. Conclusion It is suggested that high levels of TLR3 and TLR44 mRNA be are expressed in BM-MSC of healthy donors, and that the proliferation of BM-MSC isbe affected by POLY(I:C) and LPS through TLR3 and TLR4 4 .

Key words: Bone Marrow, Mesenchemal stem cell, Toll-like receptor, proliferation, bBone marrow, m Mesenchymal esenchemal stem cell, Ttoll-like receptor, Pproliferation

中图分类号: 

  • R329
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