Yang R L, Yu T T, Zhou Y H
Department of Orthodontics, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2017 Oct 18;49(5):872-877.
To analyze the role of acetylsalicylic acid (ASA) in immunomodulation of mesenchymal stem cells derived from gingiva (GMSCs), and to explore the role of ASA in enhancing the immumomodulation of GMSCs and the capacity of GMSCs to treat immune disorders and the underlying mechanism.
Flow cytometry analysis were used to analyze the role of ASA in the expression of stem cells surface markers CD146, CD105, CD90, CD34 and CD45 in GMSCs,and the GMSCs proliferation was analyzed by 5-bromo-2-deoxyuridine (BrdU) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The GMSCs and T cells co-culture system was established to analyze the role of ASA in immunomodulation of GMSCs by measuring T cell apoptosis using flow cytometry analysis and inflammatory cytokines using enzyme linked immunosorbent assay (ELISA). Further more, the dextran sulfate sodium (DSS) induced colitis mouse model was established and the mouse body weight, disease activity score, histological index and pathological change of colons were analyzed after GMSC infusion.
The proliferation of GMSCs and the expressions of CD105, CD146 in GMSCs were increased after ASA treatment. In the GMSCs and T cells co-culture system, GMSCs induced T cells apoptosis and inhibited interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) secretion by T cells, which were enhanced by ASA treatment. In vivo, GMSCs infusion could ameliorate DSS-induced colitis, including inhibited DSS-induced mouse body weight loss, decreased disease activity score and histological index, and decreased inflammation cells infiltration in colons, as shown by hematoxylin-eosin (HE) staining. Moreover, the therapeutic effects of GMSC infusion on DSS-induced colitis could be enhanced by ASA treatment. Mechanically, ASA treatment increased FasL expression of Fas/FasL death pathway in GMSCs to induce T cells apoptosis.
ASA enhanced immunomodulation of GMSCs and increased the capacity of GMSCs to ameliorate DSS-induced colitis in mice.
分析乙酰水杨酸（ASA）在牙龈间充质干细胞（GMSCs）免疫调节中的作用，探讨ASA增强GMSCs免疫调节作用及治疗免疫紊乱的能力及其潜在机制。
采用流式细胞术分析ASA对GMSCs中干细胞表面标志物CD146、CD105、CD90、CD34和CD45表达的作用，通过5-溴-2-脱氧尿苷（BrdU）染色和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法分析GMSCs的增殖情况。建立GMSCs与T细胞共培养体系，通过流式细胞术分析检测T细胞凋亡以及酶联免疫吸附测定（ELISA）检测炎性细胞因子，以分析ASA在GMSCs免疫调节中的作用。此外，建立葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型，在输注GMSCs后分析小鼠体重、疾病活动评分、组织学指标及结肠病理变化。
ASA处理后GMSCs的增殖以及GMSCs中CD105、CD146的表达增加。在GMSCs与T细胞共培养体系中，GMSCs诱导T细胞凋亡并抑制T细胞分泌干扰素γ（IFN-γ）和肿瘤坏死因子α（TNF-α），ASA处理可增强这些作用。在体内，输注GMSCs可改善DSS诱导的结肠炎，包括抑制DSS诱导的小鼠体重减轻、降低疾病活动评分和组织学指标，以及苏木精-伊红（HE）染色显示的结肠炎症细胞浸润减少。此外，ASA处理可增强输注GMSCs对DSS诱导的结肠炎的治疗效果。机制上，ASA处理增加了GMSCs中Fas/FasL死亡途径的FasL表达以诱导T细胞凋亡。
ASA增强了GMSCs的免疫调节作用，并提高了GMSCs改善小鼠DSS诱导的结肠炎的能力。
