Effects of FoxO3a on M2 polarization and ferroptosis levels of macrophages in allergic rhinitis
10.13431/j.cnki.immunol.j.20240116
- VernacularTitle:FoxO3a影响过敏性鼻炎中巨噬细胞M2型极化及铁死亡水平
- Author:
Qinghui LI
1
;
Yongliang ZENG
1
;
Xuemei CHEN
1
;
Bin XIAO
1
;
Shujun YI
1
Author Information
1. 402260,重庆大学附属江津医院耳鼻咽喉头颈外科
- Publication Type:Journal Article
- Keywords:
Allergic rhinitis;
M2 macrophage;
FoxO3a;
Ferroptosis
- From:
Immunological Journal
2024;40(11):831-838
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of Forkhead box protein O3a(FoxO3a)on M2 polarization and ferroptosis of macrophages in allergic rhinitis(AR).Methods Nasal lavage fluids were collected from 30 patients with AR and 30 healthy volunteers,while nasal mucosa samples were obtained from 3 AR patients and 3 healthy volunteers.The levels of FoxO3a in nasal lavage fluids were detected by ELISA.M0 macrophages were transfected with siRNA negative control plasmid(si-Con),siRNA FoxO3a plasmid(si-FoxO3a),pcDNA3.1-vector,or pcDNA3.1-FoxO3a plasmid,and then induced to polarize towards M2 type by interleukin-4(IL-4).M0 macrophages were set as the control group,IL-4 model group,IL-4+si-Con group,IL-4+si-FoxO3a group,IL-4+si-FoxO3a+nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor ML385 group,IL-4+pcDNA3.1-vector group,and IL-4+pcDNA3.1-FoxO3a group.A co-culture system of M0 macrophages and human nasal epithelial cells(NECs)was established,and then divided into the NECs+M0 macrophages(control group),NECs+M2 macrophages(IL-4 model group),NECs+si-FoxO3a-transfected macrophages(IL-4+si-FoxO3a group)and NECs+si-FoxO3a-transfected macrophages+ML385(IL-4+si-FoxO3a+ML385 group).Apoptosis of NECs was detected by flow cytometry.C57BL/6 mice were divided into normal group,AR model group and shRNA-FoxO3a adeno-associated virus(AAV-sh-FoxO3a)group.HE staining was used to observe histopathological changes in nasal mucosa tissue,and flow cytometry was used to detect the proportion of M2 macrophages in mouse nasal lavage fluids.The levels of IL-5,IL-13 and NECs mucin 5AC(MUC5AC)in mouse nasal lavage fluids were detected by ELISA;Western blot was used to detect the levels of FoxO3a,IL-10,arginase 1(Arg-1),Nrf2,heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4).Results Compared with the normal,the expression of FoxO3a was increased in nasal lavage fluids and nasal mucosa tissues of AR patients.Compared with the control group,the proportion of M2 macrophages and the levels of FoxO3a were increased in the IL-4 model group,while the expression levels of Nrf-2,HO-1 and GPX4 were decreased,and the levels of Fe2+and malondialdehyde(MDA)were increased(P<0.05).Compared with the IL-4 model group,these indicators were reversed in the IL-4+si-FoxO3a group.Compared with the IL-4 model group,the proportion of M2 macrophages was increased in the IL-4+pcDNA3.1-FoxO3a group.In the co-culture system,compared with the IL-4 model group,the apoptosis of NECs and MUC5AC secretion were decreased in the IL-4+si-FoxO3a group(P<0.05).ML385 reversed the protective effect of si-FoxO3a on NECs.Mice in the AAV-sh-FoxO3a group showed reduced nasal mucosa tissue damage,decreased levels of IL-5 and IL-13 in nasal lavage fluids,and a lower number of M2 macrophages,decreased level of FoxO3a in M2 macrophages,while increased expression of GPX4,Nrf2 and HO-1.Conclusion Interference with FoxO3a protein expression can inhibit macrophage ferroptosis,suppress M2 polarization,and reduce NECs damage,which is related to the activation of the Nrf-2/HO-1 signaling pathway in macrophages.
