Saikosaponin A affects differentiation and function of M1/M2 macrophages by regulating NF-κB and STAT6 signaling pathways
10.16016/j.2097-0927.202506038
- VernacularTitle:柴胡皂苷A通过调控NF-κB和STAT6信号通路影响M1/M2型巨噬细胞的分化和功能
- Author:
Zhao CUI
1
;
Yating SHU
;
Fan LEI
;
Meiyu PENG
Author Information
1. 山东第二医科大学
- Keywords:
saikosaponin A;
bone marrow derived macrophages;
cell differentiation;
NF-κB;
signal transducer and activator of transcription 6
- From:
Journal of Army Medical University
2025;47(18):2197-2210
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Saikosaponin A(SSA)on the differentiation,apoptosis and function of mouse bone marrow derived macrophages(BMDM)-derived M1/M2 macrophages,and to explore its molecular mechanism.Methods BMDM was induced to differentiate into M1/M2 macrophages in vitro,and SSA was added at the same time:CCK-8 assay was used to detect the viability of BMDM and M1/M2 macrophages.The morphology of M1/M2 macrophages was observed by inverted fluorescence microscopy.Flow cytometry(FCM)and ELISA were used to detect the levels of surface markers and cytokines in M1/M2 macrophages.Real-time fluorescent quantitative PCR(qPCR)was used to detect the mRNA levels of IL-6,TNF-α and arginase-1(Arg-1).FCM was used to detect the phagocytosis of peritoneal macrophages to fluorescent microsphere particles.Immunofluorescence(IF)assay and Western blotting were used to detect the molecular mechanism of SSA regulating M1/M2 macrophages.Results No significant effect on viability of M1/M2 macrophages was observed at SSA concentration of 10.0 mg/L,and obvious inhibition was seen at a concentration of 15.0 mg/L(P<0.01).Treatment of 10.0 mg/L SSA induced obvious morphologic changes in M1/M2 macrophages,with M1 macrophages in irregular shape,a few having pseudopods,and some showing unclear boundaries;while some M2 macrophages presenting round or irregular(P<0.001)with unclear boundaries.SSA treatment also resulted in significantly decreased proportion of M1/M2 macrophages after BMDM differentiation(P<0.05),with reduced contents of IL-6 and TNF-α secreted by M1 macrophages and their mRNA levels(P<0.05),but increased secretion of Arg-1 and mRNA levels by M2 macrophages(P<0.05).SSA treatment also inhibited the phagocytosis ability of peritoneal macrophages to fluorescent microsphere particles(P<0.01)in a concentration-dependent manner.SSA decreased the phosphorylation of NF-kappaB(p-NF-κB)(P<0.01)and enhanced the phosphorylation of signal transducer and activator of transcription 6(p-STAT6)in M2 macrophages(P<0.05).Conclusion SSA may affect the differentiation and function of M1/M2 macrophages by regulating NF-κB and STAT6 signaling pathways.
