临床输血与检验2025，Vol.27Issue(6)：782-788,7.DOI:10.3969/j.issn.1671-2587.2025.06.006
基于血小板输注体外细胞模型探究ASIC3驱动Ca2+-CaN-NFAT信号轴诱发Treg/Th17免疫失衡的作用
Role of ASIC3-driven Ca2+-CaN-NFAT Signaling Axis in Inducing Treg/Th17 Immune Imbalance Platelet Transfusion in Vitro Cell Model
摘要
Abstract
Objective To investigate the regulatory mechanism underlying the effect of acid-sensing ion channel 3(ASIC3)on the balance between regulatory T(Treg)cells and T helper type 17(Th17)cells via the Ca2+-calcineurin(CaN)-nuclear factor of activated T cells(NFAT)signaling axis.Methods Donor platelets and recipient peripheral blood mononuclear cells(PBMCs)were co-cultured in vitro at a ratio of 250∶1 to simulate the state of platelet transfusion in vitro.By constructing ASIC3 overexpression lentivirus and co-culturing with in vitro cell model,they were divided into three groups:(1)blank control group;(2)negative control lentivirus group;(3)ASIC3 overexpression lentivirus group.The proportion of Treg(Foxp3+)and Th17(IL-17A+)cells and intracellular Ca2+level were detected by flow cytometry.The expression of ASIC3,phospholipase CB(PLCB),CaN,NFATC1 and its phosphorylated protein(p-NFATC 1)were detected by Western blot.Results Compared with the blank control group,the proportion of Th17 cells in the ASIC3 overexpression lentivirus group was significantly increased(P＜0.01),and the proportion of Treg cells was significantly decreased(P＜0.05).Intracellular Ca2+concentration increased 1.8 times(P＜0.001).Protein detection showed that the expression of PLCB,CaNand p-NFATC1 in ASIC3 overexpression group were significantly up-regulated(all P＜0.05).Conclusion ASIC3 overexpression up-regulates the expression of PLCB/CaN/p-NFATC1 by activating the Ca2+-CaN-NFAT signaling axis,promotes the continuous activation of NFAT,and then drives IL-17 transcription,eventually leading to Treg/Th17 immune imbalance.关键词
酸敏离子通道/细胞实验/Treg/Th17平衡/钙离子/外周血单个核细胞Key words
ASIC3/Cell experiment/Treg/Th17 balance/Calcium ion/PBMCs分类
医药卫生引用本文复制引用
梁静,范娜,郑淑贤,刘雯..基于血小板输注体外细胞模型探究ASIC3驱动Ca2+-CaN-NFAT信号轴诱发Treg/Th17免疫失衡的作用[J].临床输血与检验,2025,27(6):782-788,7.基金项目
本课题受新疆维吾尔自治区自然科学基金面上项目(No.2022D01C585)、新疆维吾尔自治区自然科学基金青年基金项目(No.2022D01C822)资助 （No.2022D01C585）
