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低剂量X射线照射对活化巨噬细胞的影响及其可能的信号通路。

Effects of low-dose X-ray irradiation on activated macrophages and their possible signal pathways.

作者信息

Li Jian, Yao Zhen-Yu, She Chang, Li Jian, Ten Bin, Liu Chang, Lin Shu-Bin, Dong Qi-Rong, Ren Pei-Gen

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.

Department of Translational Medicine R&D Center, Shenzhen Institute of Advanced Technology, CAS, Shenzhen, Guangdong, China.

出版信息

PLoS One. 2017 Oct 27;12(10):e0185854. doi: 10.1371/journal.pone.0185854. eCollection 2017.

Abstract

Low-dose irradiation (LDI) has been used in clinics to treat human diseases, including chronic inflammation. This study assessed the effects of LDI on the inflammatory response of activated mouse primary peritoneal macrophages, and the underlying signal pathways. Primary peritoneal macrophages were isolated from mice and then incubated with lipopolysaccharide (LPS)-coated Ti microparticles (Ti-positive control) with or without brief exposure to LDI (X-ray, 0.5 Gy) 1 h later (Ti-LDI group) or left untreated in culture medium (Ti-negative control). The macrophages were then subjected to qRT-PCR, Western blot, cell viability CCK-8 assay, and ELISA. qRT-PCR analysis revealed the Ti-LDI group expressed significantly lower levels of IL-1β, IL-6, and TNF-α mRNA than those of the Ti-positive control group, while the ELISA data showed that Ti-LDI group had significantly lower secretion of IL-1β, IL-6, and TNF-α proteins. The most significant reduction associated with LDI was the secretion TNF-α protein, which barely increased from 13 to 25 h after treatment. Western blot data demonstrated that phosphorylation of p65 and ERK was much lower in the Ti-LDI group than in the controls. The data from the current study suggests that LDI of activated mouse macrophages was associated with significantly lower inflammation responses, compared with non-exposed activated macrophages, which was possibly through inhibition of the NF-κB and ERK pathways.

摘要

低剂量辐射(LDI)已在临床上用于治疗包括慢性炎症在内的人类疾病。本研究评估了LDI对活化的小鼠原代腹腔巨噬细胞炎症反应及其潜在信号通路的影响。从小鼠中分离出原代腹腔巨噬细胞,然后与包被脂多糖(LPS)的钛微粒(钛阳性对照)一起孵育,1小时后对其进行或不进行短暂的LDI照射(X射线,0.5 Gy)(钛-LDI组),或在培养基中不进行处理(钛阴性对照)。然后对巨噬细胞进行qRT-PCR、蛋白质免疫印迹、细胞活力CCK-8检测和酶联免疫吸附测定(ELISA)。qRT-PCR分析显示,钛-LDI组中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)mRNA的表达水平明显低于钛阳性对照组,而ELISA数据表明,钛-LDI组中IL-1β、IL-6和TNF-α蛋白的分泌明显减少。与LDI相关的最显著降低是TNF-α蛋白的分泌,其在治疗后13至25小时几乎没有增加。蛋白质免疫印迹数据表明,钛-LDI组中p65和细胞外信号调节激酶(ERK)的磷酸化水平远低于对照组。本研究数据表明,与未暴露的活化巨噬细胞相比,活化的小鼠巨噬细胞的LDI与明显更低的炎症反应相关,这可能是通过抑制核因子κB(NF-κB)和ERK信号通路实现的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79e7/5659615/706b324542ad/pone.0185854.g001.jpg

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