感染、炎症、修复2026，Vol.27Issue(2)：143-151,9.DOI:10.3969/j.issn.1672-8521.2026.02.007
海藻酸钠/乙二醇壳聚糖双网络水凝胶通过诱导自噬增强CD4+CD25+调节性T细胞免疫抑制活性的研究
Enhancement of CD4+CD25+regulatory T cell immunosuppressive activity via autophagy induction by sodium alginate/glycol chitosan double-network hydrogels
摘要
Abstract
Objective To investigate the regulatory effects of sodium alginate/glycol chitosan(Alg/GC)double-network hydrogels on the function of CD4+CD25+regulatory T cells(Tregs)and to elucidate the key intracellular mechanisms underlying these effects.Methods Sodium alginate/glycol chitosan(Alg/GC)double-network hydrogels were prepared.Tregs were exposed to phosphate buffered saline(PBS,PBS group),Alg/GC double-network hydrogels(Alg/GC group),rapamycin(rapamycin group)and Alg/GC+3-methyladenine(3-MA,Alg/GC+3-MA group)for 24 hours.Expressions of forkhead wing helix transcription factor and cytotoxic T lymphocyte antigen 4(CTLA-4),as well as the proliferation activity of CD4+CD25-effector T cells were measured by flow cytometry.Enzyme linked immunosorbent assay(ELISA)was used to determine the cytokine levels including interferon-γ(IFN)-γ,interleukin(IL)-10,IL-2,IL-4,and transforming growth factor β(TGF)-β.The expression of autophagy-associated proteins and formation of autophagosomes were evaluated by Western blot,transmission electron microscope and laser scanning confocal microscopy,respectively.Results After treatment with the Alg/GC double-network hydrogels,the expression levels of LC3-Ⅱ and Beclin1 in the Alg/GC group and the Rapamycin group were significantly higher than those in the PBS control group,while the expression level of P62 was significantly lower than that in the PBS control group(P<0.05).The distribution of endogenous LC3-Ⅱ in Tregs was detected by laser scanning confocal microscope,and the number of LC3 puncta in the Alg/GC group and the Rapamycin group was significantly higher than that in the PBS control group(P<0.05).Transmission electron microscope observed obvious autophagosomes in Tregs.In the experiment assessing the immunosuppressive function of Tregs,the expression of CTLA-4 and the secretion level of TGF-β in Tregs were significantly decreased in the Alg/GC+3-MA group and the Beclin1 knockout group compared with the control group,respectively(P<0.05).The inhibitory effect of Tregs on the proliferation activity of CD4+CD25-effector T cells was significantly weakened compared with the control group(P<0.05).Furthermore,the ratio of IL-4/IFN-γ secreted by CD4+CD25-effector T cells after co-culture was significantly reduced compared with the control group(P<0.05).Conclusions The Alg/GC double-network hydrogels can significantly activate the autophagic activity of Tregs.The upregulation of autophagic activity may be one of the core mechanisms mediating the enhancement of the immunosuppressive function of Tregs by Alg/GC double-network hydrogels.关键词
调节性T细胞/海藻酸钠/乙二醇壳聚糖/自噬/免疫调控Key words
regulatory T cells/sodium alginate/glycol chitosan/autophagy/immune regulation引用本文复制引用
陈坤威,潘申奥,潘丽娜,姚咏明..海藻酸钠/乙二醇壳聚糖双网络水凝胶通过诱导自噬增强CD4+CD25+调节性T细胞免疫抑制活性的研究[J].感染、炎症、修复,2026,27(2):143-151,9.基金项目
国家重点研发计划项目(2202YFA1104600) （2202YFA1104600）
温州市科技局项目(Y20210710) （Y20210710）
