新医学2025，Vol.56Issue(4)：345-353,9.DOI:10.12464/j.issn.0253-9802.2024-0475
胸腺素β4在RAW264.7巨噬细胞炎症模型中的抗炎作用机制
The anti-inflammatory mechanism of thymosin β4 in a RAW 264.7 macrophage inflammation model
摘要
Abstract
Objective To explore the effects of Thymosin β4(Tβ4)on the lipopolysaccharide(LPS)-induced polarization tendency of murine monocyte-macrophage RAW264.7 cells and its influence on inflammatory responses.Methods An inflammation model was established by LPS induction in RAW264.7 cells.Cell viability was assessed using the CCK-8 assay.The concentrations of cytokines[tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)]were detected by enzyme-linked immunosorbent assay(ELISA).Nitric oxide(NO)secretion in the cell culture supernatant was detected using the Griess method.Real-time quantitative polymerase chain reaction(RT-qPCR)was employed to detect mRNA expression levels of nuclear factor-κB(NF-κB),cyclooxygenase-2(COX-2),TNF-α,and IL-6.Western blotting was used to analyze protein levels of inducible nitric oxide synthase(iNOS),NF-κB,phosphorylated NF-κB(p-NF-κB),NF-κB inhibitor α(IκB-α),and phosphorylated IκBα(p-IκB-α).The polarization status of macrophages was observed using double fluorescence staining.Immunofluorescence staining was performed to examine NF-κB localization and expression.Results After treatment with 1 000 μg/L Tβ4 for 24 hours,the cell viability of RAW 264.7 cells was 90.2%,showing a statistically significant difference compared to the blank control group(P<0.05).Tβ4 at various concentrations effectively inhibited NO production(all P<0.000 1).Tβ4 decreased the concentrations of pro-inflammatory cytokines(TNF-α and IL-6)and NO in the LPS-induced RAW264.7 inflammatory model.It also downregulated mRNA expression of NF-κB,COX-2,TNF-α,and IL-6,as well as protein expression of iNOS,p-NF-κB,and p-IκBα.Additionally,Tβ4 inhibited NF-κB nuclear translocation and decreased CD80 expression(all P<0.05).Conclusion Tβ4 exhibits anti-inflammatory effects,the mechanisms of which may be associated with the inhibition of the NF-κB signaling pathway and suppression of macrophage M1 polarization.关键词
胸腺素β4/脓毒症/巨噬细胞/感染/炎症/NF-κB通路Key words
Thymosin β4/Sepsis/Macrophages/Infection/Inflammation/NF-κB pathway引用本文复制引用
吴天羽,林昱君,林笑宇,朱轶,余慕雪,刘王凯..胸腺素β4在RAW264.7巨噬细胞炎症模型中的抗炎作用机制[J].新医学,2025,56(4):345-353,9.基金项目
广东省自然科学基金(2022A1515010031) （2022A1515010031）
