干扰素γ对人脐带间充质干细胞生物学特性及免疫调节功能的影响  

Effects of Interferon-γ on Biological Characteristics and Immunomodulatory Property of Human Umbilical Cord-derived Mesenchymal Stem Cells

作　　者：戎丽娟 池颖 杨少光 陈丹丹 陈芳 徐淑霞 张冬雷 马凤霞 卢士红 韩忠朝 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院,实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2012年第2期421-426,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(30900557);国家重大科学计划“973计划”(2011CB964800);天津市应用基础及前沿技术研究计划(09JCYBJC09700)

摘　　要：本研究旨在探讨IFN-γ对人脐带间充质干细胞(hUC-MSC)生物学特性及免疫调节功能的影响。应用流式细胞术检测hUC-MSC表面分子表达,CCK-8检测其增殖能力,诱导hUC-MSC成脂、成骨分化并染色,ELISA检测PGE-2的分泌量,实时定量PCR方法检测hUC-MSC细胞中COX-2,IDO-1和IDO-2的表达情况,同时将IFN-γ刺激后的hUC-MSC与人外周血单个核细胞(hPBMC)共培养,检测hPBMC的增殖情况。IFN-γ组hUC-MSC以IFN-γ10ng/ml刺激24 h,同时以未刺激的hUC-MSC作为对照。结果显示:IFN-γ刺激后hUC-MSC表面SSEA-4表达量与对照组相比显著下降〔(8.15±2.94)%vs(16.42±8.5)%,P<0.05〕,CD54表达升高〔(96.64±3.29)%vs(84.12±10.73)%,P=0.051〕。IFN-γ可以增强hUC-MSC对活化的hPBMC的增殖抑制能力(P<0.05),具有浓度依赖性。IFN-γ刺激24 h后,hUC-MSC分泌PGE-2明显降低(P<0.01),COX-2表达变化与对照组相比无统计学意义,但具有降低趋势;IDO-1的表达与对照组相比显著增高(P<0.01),IDO-2表达无明显变化。结论:IFN-γ刺激影响hUC-MSC表面分子表达及分化等基本生物学特性,并增强hUC-MSC的免疫调节功能。The aim of this study was to investigate the effects of interferon（IFN）-γ on biological characteristics and immunomodulatory property of human umbilical cord-derived mesenchymal stem cells（hUC-MSC）.hUC-MSC were treated with IFN-γ 10 ng/ml（IFN-γ group） or without IFN-γ （control group）.The phenotype of hUC-MSC was detected by flow cytometry.The proliferation status was detected by CCK-8 method,and its differentiation ability was assessed by oil red O and von Kossa staining.The production of PGE-2 was measured by ELISA,and the mRNA expression levels of COX-2,IDO-1 and IDO-2 in hUC-MSC were detected by real-time quantitative PCR.Furthermore,the proliferation of human peripheral blood mononuclear cells（hPBMNC） was evaluated after co-culture with hUC-MSC,IFN-γ pretreatment or not.The results showed that after IFN-γ stimulation,the expression of SSEA-4 on hUC-MSC decreased significantly（（8.15±2.94）% vs（16.42±8.5）%,P0.05）,and the expression of CD54 increased（（96.64±3.29）% vs（84.12±10.73）%,P=0.051）.The immunomodulatory property of hUC-MSC on the proliferation of hPBMNC was enhanced（P0.05）.All the above mentioned effects were IFN-γ concentration-dependent.When hUC-MSC were stimulated by IFN-γ for 24 h,the production of PGE-2 secreted by hUC-MSC decreased significantly（P0.01）.The mRNA expression level of COX-2 also decreased though the difference did not reach to statistically significant level.Compared with control group,IDO-1 expression level in IFN-γ group increased significantly（P0.01）,and the mRNA expression level of IDO-2 remained unchanged.It is concluded that IFN-γ can influence the phenotype of hUC-MSC and enhance the immunomodulatory property of hUC-MSC.

关 键 词：人脐带间充质干细胞 干扰素Γ 生物学特性 免疫调节 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R979.5[医药卫生—基础医学]