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  

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

作　　者：赵强[1] 张馥敏[1] 王连生[1] 王昭军[1] 马文珠[1] 

机构地区：[1]南京医科大学第一附属医院心内科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2009年第3期345-349,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：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)

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、20.0μg/mL)分别与野生型小鼠MSCs共培养24h和48h,采用CellCountKit-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LPS1.0μg/ml共培养,探讨LPS促进MSCs增殖的可能机制。结果:LPS(0.1～5.0μg/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5.0μg/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LPS20.0μg/ml组增殖能力仍高于对照组(P>0.05);LPS对LR4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。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. Flowcy- tometry 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 μg/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 μg/ml were used to explore the possible mechanism of promoting MSCs proliferation. Results： Low dose LPS（0.1-5.0 μg/ml） strikingly improved the proliferation of MSCs in vitro （P 〈 0.05）. The stimulation with LPS 5.0μg/ml reached maximum effects on MSCs proliferation. While the concentration of LPS was further increased more than 5.0 μg/ml, the improvement effects on MSCs proliferation showed a tendency to decline, but the effect at 20.0 μg/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

关 键 词：骨髓间充质干细胞 脂多糖 TOLL样受体4 增殖 

分 类 号：R331.2[医药卫生—人体生理学]