米诺环素可能通过靶向 SOCS1/TLR4/NF-κB 信号通路诱导树突状细胞产生耐受。
Minocycline induces tolerance to dendritic cell production probably by targeting the SOCS1/ TLR4/NF-κB signaling pathway.
机构信息
Department of Neurology, Key Laboratory of Neurology of Hebei Province, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, People's Republic of China.
Department of Neurology, Key Laboratory of Neurology of Hebei Province, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, People's Republic of China.
出版信息
Transpl Immunol. 2023 Aug;79:101856. doi: 10.1016/j.trim.2023.101856. Epub 2023 May 16.
OBJECTIVE
Dendritic cells (DCs) are professional antigen-presenting cells that play a key role in maintaining peripheral immune tolerance. The use of tolerogenic DCs (tolDCs), i.e., semi-mature DCs that express co-stimulatory molecules but not pro-inflammatory cytokines, has been proposed. However, the mechanism of tolDCs induced by minocycline is still unclear. Our previous bioinformatics analyses based on multiple databases suggested that the suppressor of cytokine signaling 1/Toll-like receptor 4/NF-κB (SOCS1/TLR4/NF-κB) signal pathway was associated with DCs maturation. Thus, we studied whether minocycline could induce DC tolerance through this pathway.
METHODS
A search for potential targets was carried out through public databases, and pathway analysis was performed on these potential targets to obtain pathways relevant to the experiment. Flow cytometry was used to detect the expression of DC surface markers CD11c, CD86, and CD80, and major histocompatibility complex II. The secretion of interleukin (IL)-12p70, tumor necrosis factor alpha (TNF- α), and IL-10 in the DC supernatant was detected by enzyme-linked immunoassay. The ability of three groups (Ctrl-DCs, Mino-DCs, and LPS-DCs) of DCs to stimulate allogeneic CD4+ T cells was analyzed using a mixed lymphocyte reaction assay. Western blotting was used to detect the expression of TLR4, NF-κB-p65, NF-κB-p-p65, IκB-α, and SOCS1 proteins.
RESULTS
The hub gene plays a vital role in biological processes; in related pathways, the regulation of other genes is often affected by it. The SOCS1/TLR4/NF-κB signaling pathway was further validated by searching for potential targets through public databases to obtain relevant pathways. The minocycline-induced tolDCs showed characteristics of semi-mature DCs. Moreover, the IL-12p70 and TNF-α levels in the minocycline-stimulated DC group (Mino-DC group) were lower than those in the lipopolysaccharide (LPS)-DC group, and the IL-10 levels were higher in the Mino-DC group than in the LPS-DC and control DC groups. In addition, the Mino-DC group had decreased protein expression levels of TLR4 and NF-κB-p65 and upregulated protein levels of NF-κB-p-p65, IκB-α, and SOCS1 compared with the other groups.
CONCLUSION
The results of this study indicate that minocycline could improve the tolerance of DCs probably by blocking the SOCS1/TLR4/NF-κB signaling pathway.
目的
树突状细胞（DCs）是一种专业的抗原呈递细胞，在维持外周免疫耐受中起着关键作用。已经提出使用致耐受性 DC（tolDCs），即表达共刺激分子但不表达促炎细胞因子的半成熟 DC。然而，米诺环素诱导 tolDC 的机制尚不清楚。我们之前基于多个数据库的生物信息学分析表明，细胞因子信号转导抑制因子 1/Toll 样受体 4/核因子-κB（SOCS1/TLR4/NF-κB）信号通路与 DC 成熟有关。因此，我们研究了米诺环素是否可以通过该途径诱导 DC 耐受。
方法
通过公共数据库进行潜在靶标的搜索，并对这些潜在靶标进行通路分析，以获得与实验相关的通路。流式细胞术检测 DC 表面标志物 CD11c、CD86 和 CD80 以及主要组织相容性复合体 II 的表达。酶联免疫吸附试验检测 DC 上清液中白细胞介素（IL）-12p70、肿瘤坏死因子-α（TNF-α）和 IL-10 的分泌。混合淋巴细胞反应试验分析三组 DC（Ctrl-DCs、Mino-DCs 和 LPS-DCs）刺激同种异体 CD4+T 细胞的能力。Western blot 检测 TLR4、NF-κB-p65、NF-κB-p-p65、IκB-α和 SOCS1 蛋白的表达。
结果
枢纽基因在生物学过程中起着至关重要的作用；在相关通路中，它往往会影响其他基因的调控。通过公共数据库搜索潜在靶点进一步验证了 SOCS1/TLR4/NF-κB 信号通路，获得了相关通路。米诺环素诱导的 tolDC 具有半成熟 DC 的特征。此外，米诺环素刺激的 DC 组（Mino-DC 组）中 IL-12p70 和 TNF-α水平低于脂多糖（LPS）-DC 组，而 Mino-DC 组中 IL-10 水平高于 LPS-DC 和对照 DC 组。此外，与其他组相比，Mino-DC 组 TLR4 和 NF-κB-p65 的蛋白表达水平降低，NF-κB-p-p65、IκB-α和 SOCS1 的蛋白表达水平升高。
结论
本研究结果表明，米诺环素可能通过阻断 SOCS1/TLR4/NF-κB 信号通路来提高 DC 的耐受性。
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