丹皮酚对牙龈卟啉单胞菌诱导骨髓来源巨噬细胞功能的影响

发布时间：2018-03-15 20:34

本文选题：丹皮酚　切入点：牙龈卟啉单胞菌　出处：《华西口腔医学杂志》2017年02期 　论文类型：期刊论文

【摘要】：目的观察丹皮酚对牙龈卟啉单胞菌作用下对体外培养的小鼠骨髓来源巨噬细胞(BMM)炎性因子分泌及向破骨细胞分化能力的影响,并探讨其作用机制。方法体外分离获得小鼠生长良好的BMM,加入不同浓度丹皮酚溶液处理1 h,再加入牙龈卟啉单胞菌进行24 h诱导刺激。采用流式细胞术检测炎症相关蛋白程序性死亡分子配体1(PD-L1)的表达量,采用酶联免疫吸附试验(ELISA)检测细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及白细胞介素-6(IL-6)水平;在核因子κB受体活化因子配体(RANKL)和巨噬细胞集落刺激因子(M-CSF)刺激后的BMM中加入不同浓度的丹皮酚溶液处理1 h后,加入牙龈卟啉单胞菌刺激,采用抗酒石酸性磷酸酶(TRAP)染色观察破骨细胞形成情况,Western blot检测破骨细胞形成相关蛋白TRAP和核因子κB受体活化因子(RANK)的表达。结果丹皮酚在10~50μmol·L-1的范围内,对BMM无毒性作用。流式细胞术结果提示丹皮酚可以抑制牙龈卟啉单胞菌诱导PD-L1的表达,并存在剂量依赖效应。ELISA实验证明丹皮酚能以剂量依赖性方式抑制BMM释放TNF-α、IL-1β及IL-6(P0.01)。牙龈卟啉单胞菌可以明显诱导BMM分化形成破骨细胞;TRAP染色显示丹皮酚各浓度组能抑制BMM向破骨细胞分化;Western blot结果显示丹皮酚抑制TRAP和RANK表达且存在剂量依赖效应。结论丹皮酚可以剂量依赖方式抑制牙龈卟啉单胞菌诱导的BMM炎性因子TNF-α、IL-1β及IL-6的分泌及其向破骨细胞分化的能力。
[Abstract]:Objective to observe the effect of paeonol on the secretion of inflammatory cytokines and differentiation ability into osteoclasts of murine bone marrow-derived macrophages (BMMs) induced by porphyromonas gingivalis in vitro. Methods BMMs with good growth were isolated from mice in vitro, treated with different concentrations of paeonol for 1 hour, and then induced by porphyromonas gingivalis for 24 h. Flow cytometry was used to detect inflammation. The expression of associated protein programmed death molecular ligand 1 (PD-L1), The levels of tumor necrosis factor- 伪 (TNF- 伪), interleukin-1 尾 (IL-1 尾) and interleukin-6 (IL-6) in the supernatant of the cells were detected by enzyme-linked immunosorbent assay (Elisa). BMM stimulated by nuclear factor- 魏 B receptor activating factor ligand (RANKL) and macrophage colony stimulating factor (M-CSF) were treated with different concentrations of paeonol for 1 h, then stimulated by porphyromonas gingivalis. The expression of osteoclast-associated protein (TRAP) and nuclear factor 魏 B receptor activating factor (RANKK) in osteoclasts were detected by anti-tartaric acid phosphatase (TRAP) staining. Results Paeonol was found to be in the range of 1050 渭 mol 路L ~ (-1). The results of flow cytometry showed that paeonol could inhibit the expression of PD-L1 induced by porphyromonas gingivalis. The dose-dependent effect. Elisa showed that paeonol could inhibit the release of TNF- 伪, IL-1 尾 and IL-6P0.01 by BMM in a dose-dependent manner. Porphyromonas gingivalis could induce the differentiation of BMM into osteoclasts. Trap staining showed that paeonol could inhibit the release of TNF- 伪, IL-1 尾 and IL-6. The results of Western blot showed that paeonol inhibited the expression of TRAP and RANK in a dose-dependent manner. Conclusion Paeonol can inhibit BMM inflammatory factor TNF- 伪 IL-1 尾 and IL-6 induced by Porphyromonas gingivalis in a dose-dependent manner. The ability to secrete and differentiate into osteoclasts.
【作者单位】：贵阳市口腔医院牙体牙髓病科遵义医学院附属贵阳口腔医院;浙江大学医学院附属口腔医院牙体牙髓病科;
【基金】：贵阳市高层次创新型青年科技人才培养计划[筑教发(2-012HK)209-50号] 西部地区人才培养出国特别项目[(2013)5044号]~~
【分类号】：R781.4

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