【摘要】 目的研究干扰素γ(IFN-γ)对人羊膜间充质干细胞(hAMSCs)的增殖、凋亡及相关细胞因子分泌水平的影响。方法在hAMSCs培养基中加入不同浓度IFN-γ,之后采用细胞增殖-毒性检测试剂(CCK-8)检测hAMSCs增殖情况,流式细胞术(FCM)检测48 h时hAMSCs的凋亡情况,ELISA检测上清中炎性因子可溶性白细胞抗原G(sHLA-G)、前列腺素E2(PGE-2)分泌水平和紫外分光光度法检测犬尿氨酸(KYN)代谢含量。结果随着培养时间的延长,无IFN-γ组和含不同浓度IFN-γ组中hAMSCs的增殖从第1天至第4天,均快速生长,随后生长速度减慢;FCM检测细胞凋亡结果显示,100 ng/mL IFN-γ培养组和无IFN-γ组相比,hAMSCs无明显凋亡(P=0.78);不同浓度10~100 ng/mL IFN-γ组同无IFN-γ组比较,PGE-2、sHLA-G分泌水平显著增加(P <0.05),同时吲哚胺2,3-二氧化酶(IDO)分解色氨酸的代谢产物KYN水平显著增加(P <0.05),IDO酶活性增强。结论 IFN-γ在不影响hAMSCs增殖生长的条件下,促进PGE-2、sHLA-G分泌,增强IDO酶活性,为hAMSCs防治移植物抗宿主病提供理论依据。更多还原

【Abstract】 Objective To study the effect of IFN-gamma(IFN-γ)on the proliferation,apoptosis and secretion of inflammatory factors of human amniotic mesenchymal stromal cells(hAMSCs). Methods hAMSCs were cultured with different concentrations of IFN-γ for 48 h. The proliferation of hAMSCs were determined by Cell Counting Kit-8(CCK-8). The apoptosisof hAMSCs was detected by flow cytometry(FCM). The secretion level of inflammatory factors including soluble human leukocyte antigen G(sHLA-G)and prostaglandin E2(PGE-2)were detected by ELISA assays,and kynurenine(KYN)was detected by ultraviolet spectrophotometry in the supernatant.Results With the extension of culture time,the proliferation of hAMSCs in the groups with or without IFN-γ grew rapidly from day 1 to day 4,and then the growth rate was slowed down. FCM results indicated that there was no significant apoptosis in the group treated with IFN-γ at the concentration of 100 ng/ml compared with the group treated without IFN-γ(P = 0.78). The levels of PGE-2 and sHLA-G were increased significantly in the groups treated with IFN-γcompared with the group treated without IFN-γ(P < 0.05). The level of KYN was increased significantly in the groups with IFN-γ(P < 0.001),suggesting that the activity of IDO decomposing tryptophan was strengthened. Conclusion IFN-γ has no effect on the growth and proliferation of hAMSCs,but promotes the secretionof PGE-2 and sHLA-G,and enhances the activity of IDO enzyme,providing theoretical basis for hAMSCs to prevent and treat graft-versus-host disease clinically.更多还原