Poly(I:C)影响间充质干细胞的成骨分化及其对树突状细胞的作用  

Effect of Poly(I:C) on the osteogenic differentiation and the dendritic cell-suppressive functions of MSCs

作　　者：赵晓寅[1] 刘丹[1] 刘柳[1] 侯亚义[1] 

机构地区：[1]南京大学医学院免疫与生殖生物学实验室,南京210093

出　　处：《中国免疫学杂志》2013年第1期14-19,24,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81072410);江苏省卫生厅兴卫工程基金(XK200709);江苏省第五批"六大人才高峰资助"项目资助

摘　　要：目的:间充质干细胞(MSC)能够表达Toll样受体(TLR),但TLR配体刺激对MSC的影响依然存在争议。本研究分析了TLR3配体刺激对脐带MSC表型、细胞增殖、凋亡、成骨分化和功能的影响。方法:用混合酶法分离培养了人脐带来源的MSC;用流式细胞仪分析了TLR3配体——Poly(I:C)刺激后MSC表面标志CD29、CD44、CD105、CD31、CD117和CD45表达的变化;用CCK-8和Annexin-V/PI法分别检测了不同浓度Poly(I:C)刺激24及48小时后MSC的增殖和凋亡情况;进一步分析了Poly(I:C)对MSC成骨分化的影响。此外在Poly(I:C)和MSC存在时,流式检测了DC成熟表面标志CD40、CD86、MHCⅡ的表达,DC对右旋糖酐的吞噬,混合淋巴实验检测了DC的抗原递呈能力。结果:Poly(I:C)的刺激不影响MSC的表型和增殖;只有高浓度组的Poly(I:C)刺激引起MSC的凋亡;Poly(I:C)抑制成骨相关基因RUNX2、ALP和OC的表达,抑制MSC的成骨分化;Poly(I:C)存在的情况下,MSC下调DC成熟表面标志CD40、CD86和MHCⅡ的表达,增强DC对右旋糖酐的吞噬能力,并抑制DC对T的活化作用。结论:Poly(I:C)能够影响MSC的成骨分化及免疫调节功能,提示TLR3信号的活化可能在成骨分化中发挥作用。Objective：Mesenchymal stem cells （MSCs） can express Toll-like receptors （TLRs）, but it remains unclare on the effect of TLR ligand on MSCs. Here, we studied the impact of the TLR3 ligand-Poly （I：C） on the phenotype, cell viability, apopto- sis, osteoblast differentiation and immune-suppressive functions of MSC. Methods ： The mixed enzyme method were used for the isola- tion and culture of human umbilical cord mesenchymal stem cells （MSCs）. FCM was used to examined the expression of CD29, CIM4, CD105, CD31, CD117 and CIM5 on MSC with the treatment of TLR3 ligand-Poly （I：C）. CCK-8 and Annexin-V/PI were used to measure cell viability and apoptosis with the treatment of a series of concentrations of Poly （I：C）. Then we investigated the effect of Poly（I：C） on osteoblast differentiation of MSC. FCM was used to measure the expression of CD40, CD86, MHC 1I and the cellular uptake of Dextran of DC, and the allogeneic MLR assay was used to investigate antigen-presenting function of DC both in the presence of Poly（I：C） and MSCs. Results： Poly （I：C） did not affect the phenotype and proliferation of MSC. Only high concentrations of Poly （I.C） caused apoptosis of MSC. Moreover, Poly （I.C） can inhibit the expression of RUNX2, ALP and OC in MSC. In addition, both the expression of CIM0, CD86 and MHC II were upregulated on DC, the phagocytic capacity of DC was enhanced and the activation on T cells of DC was inhibited in the presence of Poly（ I ： C） and MSCs. Conclusion： TLR3 ligand-Poly （ I ： C） could affect the differenti- ation potential and the immunol-suppressive functions of MSCs.

关 键 词：Poly(I C) 间充质干细胞 成骨分化 树突状细胞 

分 类 号：R392[医药卫生—免疫学]