FK506对干扰素-γ和肿瘤坏死因子-α诱导间充质干细胞免疫抑制的影响  

Effect of FKS06 on the immunosuppression of mesenchymal stem cells induced by interferon-γ and tumor necrosis factor-α

作　　者：唐礼功[1] 谢森[1] 陈琪[1] 李志雄[1] 

机构地区：[1]广州军区武汉总医院泌尿外科,武汉430070

出　　处：《中华实验外科杂志》2011年第6期904-906,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察用免疫抑制剂FKS06对干扰素（IFN）-γ和肿瘤坏死因子（TNF）-α诱导的间充质干细胞（MSCs）免疫抑制功能的影响及其作用机制。方法将经过不同炎症因子预处理的小鼠MSCs与脾细胞共培养，并在FKS06作用72h后，以噻唑蓝（M3T）比色法检测脾细胞增殖；以流式细胞术检测脾细胞凋亡；以实时定量聚合酶链反应（Real—timePCR）法检测MSCs发挥免疫抑制作用的主要分子，观察FKS06对其影响。结果IFN-γ和TNF-α联合预处理后的MSCs可以显著抑制脾细胞增殖，促进脾细胞凋亡，凋亡率可达（53．5±2．5）％，明显高于IFN-γ或TNF-α预处理的MSCs组以及对照组（P〈0．05），并且IFN-γ和TNF-α可以显著上调MSCs中诱导型一氧化氮合酶（iNOS）的表达，MSCs的免疫抑制作用可以被iNOS特异性抑制剂1400W所拮抗；但FKS06则可以抑制MSCs中iNOS的表达，使MSCs对脾细胞增殖的抑制作用显著降低，脾细胞凋亡减少，凋亡率降至（44．3±3．2）％（P〈0．05）。结论FK506通过抑制IFN-γ和TNF-α联合预处理的MSCs中iNOS表达从而抑制其免疫抑制功能。Objective To investigate the mechanism of immunosuppressant FKS06 on the immunesuppression of mesenchymal stem cells （MSCs） induced by interferon （IFN） -γ and tumor necrosis factor （TNF）-α. Methods MSCs pretreated with inflammatory cytokines were co-cultured with mice spleen ceils. After incubation with FKS06 for 72 h, proliferation of spleen cells was measured by methyl thiazol tetrazolium （MTT） assay and the apoptosis was assessed by flow cytometry. Real-time polymerase chain reaction （PCR） was used to detect the major molecule that mediated the immunosuppression of MSCs and the effect of FKS06 on the molecule. Results MSCs pretreated with inflammatory cytokines could inhibit the proliferation of spleen cells and increase the apoptosis of spleen cells, and the apoptosis rate was （53.5 ± 2. 5） %, which was higher than that of MSCs pretreated with IFN-γ, or TNF-α and control group （ P 〈 0. 05 ）, and IFN-γand TNF-α upregulated inducible nitric oxide synthase （iNOS） expression in MSCs significantly, and this immunosuppression could also be antagonized by a specific inhibitor 1400W. However, FKS06 could inhibit iNOS expression in MSCs pretreated with IFN-γ and TNF-α, the inhibition of spleen ceils proliferation by MSCs was reduced significantly, and the apoptosis rate of spleen ceils was reduced to （44. 3 ± 3.2 ） % （ P 〈 0. 05 ）. Conclusion FKS06 inhibits the immunosuppression of MSCs through downregulation of iNOS expression in MSCs pretreated with IFN-γ and TNF-α.

关 键 词：间充质干细胞 炎症 免疫抑制 FK506 

分 类 号：R285.5[医药卫生—中药学]