Effect of mtROS/NLRP3 signaling pathway on macrophage polarization during iron overload-induced liver fibrosis
10.3969/j.issn.1000-4718.2025.09.012
- VernacularTitle:mtROS/NLRP3信号通路对铁过载诱导的肝纤维化过程中巨噬细胞极化的影响
- Author:
Jiawen YU
1
;
Yi ZHOU
;
Chunmei QIAN
;
Lan MU
;
Renye QUE
Author Information
1. 上海中医药大学附属上海市中西医结合医院脾胃病科,上海 200082;上海中医药大学附属市中医医院消化内科,上海 200071
- Publication Type:Journal Article
- Keywords:
iron overload;
mtROS/NLRP3 signaling pathway;
hepatic fibrosis;
macrophage polarization
- From:
Chinese Journal of Pathophysiology
2025;41(9):1765-1774
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the function of mitochondrial reactive oxygen species(mtROS)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway in modulating macrophage polarization in liver fibrosis resulting from iron overload.METHODS:Thirty-two male C57BL/6 mice were randomly allocated into four groups:control group,model group(iron dextran,50 mg/kg),MitoTEMPO(3 mg/kg)group,and MCC950(10 mg/kg)group,comprising eight mice per group.All mice,with the exception of the control group,were administered daily in-traperitoneal injections of iron dextran for a duration of seven consecutive weeks,whereas the control group received equiv-alent volumes of normal saline.Starting in week four,the MitoTEMPO and MCC950 cohorts received their designated treatments through intraperitoneal injection three times weekly.Serum alanine aminotransferase(ALT)and aspartate ami-notransferase(AST)concentrations were assessed through biochemical analysis.Liver tissues were analyzed utilizing HE,Masson,Sirius red and immunohistochemical staining.The concentrations of mtROS were evaluated utilizing the MitoSOX Red probe.Cytokines and polarization markers,such as interleukin-1β(IL-1β),IL-18,IL-6,tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS),IL-10,and arginase-1(Arg-1),were quantified via ELISA.Western blot analysis was performed to quantify the protein expression levels of Arg-1,iNOS,NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),and caspase-1.The mRNA expression of NLRP3,ASC,and caspase-1 was assessed using RT-qPCR.Immunofluorescence double labeling was employed to identify M1 and M2 macrophages.RESULTS:(1)In comparison to the control group,the model group demonstrated notable inflammatory cell infiltration,pronounced fibrous tissue hyperplasia,significant disruption of hepatic lobular architecture,and the develop-ment of pseudo-lobules in certain areas.Serum ALT and AST levels were markedly elevated(P<0.01),as were the mRNA and protein expression levels of NLRP3,ASC,caspase-1,and mtROS(P<0.01).Iron overload resulted in markedly ele-vated serum iron,ferritin,total liver iron,and ferrous iron concentrations(P<0.01).Markers indicative of M1 macrophage polarization,including IL-6,TNF-α,and iNOS,exhibited upregulation(P<0.01),whereas M2 markers such as IL-10,Arg-1,and CD206 were significantly downregulated(P<0.01).(2)Compared with model group,inhibiting mtROS or NL-RP3 substantially reduced inflammation and fibrous tissue hyperplasia.ALT and AST levels were markedly diminished(P<0.01),as were the areas of positive staining for α-smooth muscle actin and collagen type Ⅰ(P<0.01).Markers of iron over-load,such as serum iron,ferritin,total liver iron,and ferrous iron,were significantly ameliorated(P<0.01).M1 polariza-tion markers were significantly downregulated(P<0.01).CONCLUSION:The mtROS/NLRP3 signaling pathway facili-tates liver fibrosis caused by iron overload by enhancing macrophage polarization to the M1 phenotype.
