Rehabilitation Medicine, NYU Grossman School of Medicine, New York, New York, U.S.A.
Department of Microbiology, NYU Grossman School of Medicine, New York, New York, U.S.A.
Laryngoscope. 2023 Nov;133(11):3116-3122. doi: 10.1002/lary.30763. Epub 2023 May 29.
The diversity of glucocorticoid (GC) properties may underlie variability of clinical efficacy for vocal fold (VF) disease. Optimized therapeutic approaches must account for tissue complexity as well as interactions between cell types. We previously reported that reduced GC concentrations inhibited inflammation without eliciting fibrosis in mono-cultured VF fibroblasts and macrophages. These data suggested that a refined approach to GC concentration may improve outcomes. In the current study, co-culture of VF fibroblasts and macrophages was employed to investigate the effects of different concentrations of methylprednisolone on fibrotic and inflammatory response genes in VF fibroblasts to optimize management paradigms.
In vitro.
THP-1 monocyte-derived macrophages were stimulated with interferon-γ (IFN-γ), lipopolysaccharide (LPS), or transforming growth factor-β (TGF-β) to induce inflammatory (M(IFN/LPS)) and fibrotic (M(TGF)) phenotypes. Macrophages were then co-cultured with a human VF fibroblast cell line using a 0.4 μm pore membrane with or without 0.1-3000 nM methylprednisolone. Inflammatory (CXCL10, TNF, and PTGS2) and fibrotic (ACTA2, CCN2, and COL1A1) gene expression was quantified in fibroblasts.
Incubating VF fibroblasts with M(IFN/LPS) macrophages increased expression of TNF and PTGS2, and this effect was inhibited by methylprednisolone. Incubation of VF fibroblasts with M(TGF) macrophages increased expression of ACTA2, CCN2, and COL1A1, and this effect was enhanced by methylprednisolone. The concentration of methylprednisolone required to downregulate inflammatory genes (TNF and PTGS2) was lower than that to upregulate fibrotic genes (ACTA2, CCN2, and COL1A1).
Reduced concentration of methylprednisolone effectively suppressed inflammatory genes without enhancing fibrotic genes, suggesting that a refined approach to GC concentration may improve clinical outcomes.
N/A Laryngoscope, 133:3116-3122, 2023.
糖皮质激素（GC）性质的多样性可能是声带（VF）疾病临床疗效差异的基础。优化的治疗方法必须考虑到组织复杂性以及细胞类型之间的相互作用。我们之前的研究报告表明，降低 GC 浓度可以在体外抑制炎症而不引起成纤维细胞和巨噬细胞纤维化。这些数据表明，精细控制 GC 浓度可能会改善结果。在目前的研究中，我们采用共培养 VF 成纤维细胞和巨噬细胞的方法，研究不同浓度甲泼尼龙对 VF 成纤维细胞中纤维化和炎症反应基因的影响，以优化管理模式。
体外。
用干扰素-γ（IFN-γ）、脂多糖（LPS）或转化生长因子-β（TGF-β）刺激 THP-1 单核细胞衍生的巨噬细胞，诱导炎症（M（IFN/LPS））和纤维化（M（TGF））表型。然后将巨噬细胞与人类 VF 成纤维细胞系在 0.4μm 孔径膜上共培养，有或没有 0.1-3000nM 甲泼尼龙。在成纤维细胞中定量检测炎症（CXCL10、TNF 和 PTGS2）和纤维化（ACTA2、CCN2 和 COL1A1）基因的表达。
与 M（IFN/LPS）巨噬细胞共孵育的 VF 成纤维细胞增加了 TNF 和 PTGS2 的表达，这种作用被甲泼尼龙抑制。与 M（TGF）巨噬细胞共孵育的 VF 成纤维细胞增加了 ACTA2、CCN2 和 COL1A1 的表达，这种作用被甲泼尼龙增强。下调炎症基因（TNF 和 PTGS2）所需的甲泼尼龙浓度低于上调纤维化基因（ACTA2、CCN2 和 COL1A1）所需的浓度。
降低甲泼尼龙浓度可有效抑制炎症基因，而不增强纤维化基因，提示精细控制 GC 浓度可能改善临床结局。
无。喉科学，133：3116-3122，2023。
