Toll样受体4及其配体脂多糖对骨髓间充质干细胞增殖的影响
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江苏省科委自然科学基金(BK2007254);人事部留学人员科研择优项目(DG216D5021)


Effects on proliferation of mesenchymal stem cells induced by Toll like receptor4 and its ligand lipopolysaccharides
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    摘要:

    目的:观察不同浓度脂多糖(lipopolysaccharides,LPS)对离体小鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)增殖功能的影响-方法:分别采取野生型和Toll样受体4(toll like receptor4,TLR4)基因缺失小鼠骨髓以贴壁分离法培养骨髓间充质干细胞,取第3~6代的MSCs,通过流式细胞术检测MSCs表面标志显示CD34-CD45阴性;CD105-CD29-CD44阳性-以不同浓度LPS(0.1-1.0-5.0-10.0-20.0 -滋g/mL)分别与野生型小鼠MSCs共培养24 h和48 h,采用Cell Count Kit-8比色法观察LPS对MSCs增殖功能的影响,此外采用TLR4基因缺失小鼠MSCs与LPS 1.0 -滋g/ml共培养,探讨LPS促进MSCs增殖的可能机制-结果:LPS(0.1~5.0 -滋g/ml)能显著提高离体野生型小鼠MSCs的增殖能力(P < 0.05),LPS浓度在5.0 -滋g/ml时对MSCs增殖功能影响最大,随着LPS浓度的继续增大,MSCs的增殖功能反呈下降趋势,但LPS 20.0 -滋g/ml组增殖能力仍高于对照组(P > 0.05);LPS对LR 4基因缺失小鼠MSCs增殖能力无显著影响-结论:适量的LPS能增加MSCs的增殖能力,机制可能是通过激活Toll样受体信号通路实现的-

    Abstract:

    Objective:To observe the effects of lipopolysaccharides(LPS) of different concentrations on the proliferation of mesenchymal stem cells(MSCs). Methods:Mesenchymal stem cells(MSCs) were isolated from wild and TLR4 gene deletion mice respectively by method of differential adherence to plastic, and their phenotypical properties were analyzed in passage 3-6 by flowcytometer. Flowcytometry showed that MSCs were CD34-,CD45- and CD105+-CD29+-CD44+. MSCs isolated from wild mice were treated with LPS of different concentrations(0.1,1.0,5.0,10.0,20.0 -滋g/ml). MSCs proliferation was detected by CCK-8(Cell Count Kit-8) co-cultured method at 24 h and 48 h respectively. In addition, MSCs isolated from TLR4 gene deletion mice incorpted with LPS 1.0 -滋g/ml were used to explore the possible mechanism of promoting MSCs proliferation. Results:Low dose LPS(0.1~5.0 -滋g/ml) strikingly improved the proliferation of MSCs in vitro(P < 0.05). The stimulation with LPS 5.0 -滋g/ml reached maximum effects on MSCs proliferation. While the concentration of LPS was further increased more than 5.0 -滋g/ml, the improvement effects on MSCs proliferation showed a tendency to decline, but the effect at 20.0 -滋g/ml was still greater than those in control group(P > 0.05). There was no significant effects on the proliferation of MSCs isolated from TLR4 gene deletion mice induced by LPS. Conclusion:The present study revealed that LPS could improve the proliferation of MSCs through TLR4, and the mechanism might activate the Toll -like receptor4 pathway

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赵 强,张馥敏,王连生,王昭军,马文珠. Toll样受体4及其配体脂多糖对骨髓间充质干细胞增殖的影响[J].南京医科大学学报(自然科学版),2009,29(3):345-349

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  • 收稿日期:2008-07-29
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