Salivary pellets induce a pro-inflammatory response involving the TLR4-NF-kB pathway in gingival fibroblasts.
Salivary pellets induce a pro-inflammatory response involving the TLR4-NF-kB pathway in gingival fibroblasts.
复制标题
唾液颗粒会引起伴随牙龈成纤维细胞中TLR4-NF-KB途径的促炎反应。
DOI：
10.1186/s12903-016-0229-5
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发表时间：
2016-07-08
期刊：
影响因子：
2.9
通讯作者：
Gruber RR
中科院分区：
文献类型：
--
作者：
Müller HD;Cvikl BB;Lussi AA;Gruber RR
全唾液在牙龈成纤维细胞中引起大量促炎反应。这就提出了一个问题，即用于诊断目的的唾液颗粒是否也具有促炎能力，如果是的话，分子水平上的潜在机制是什么。我们通过实时聚合酶链反应和免疫分析检测了广泛洗涤的唾液颗粒激发牙龈成纤维细胞中趋化因子表达的能力。用蛋白质组学分析确定蛋白质组成。通过鲎试验分析内毒素，并通过亲和色谱法除去内毒素。抑制剂TAK-242和BAY 11 -7082分别用于确定TLR 4和NF-kB信号传导的参与。Western blot检测磷酸化p65。实验表明，唾液颗粒和相应的洗涤溶液含有促炎活性而不损害细胞活力。蛋白质组学分析显示蛋白质与脂多糖的结合能力，鲎试验表明唾液颗粒中存在内毒素。用TAK-242阻断TLR 4和清除内毒素均降低了唾液颗粒增加趋化因子表达和p65磷酸化的能力。BAY 11 -7082抑制响应于唾液颗粒的趋化因子表达。高压灭菌的唾液颗粒也降低了它们的促炎活性。这些数据支持暴露于洗涤唾液颗粒制剂的牙龈成纤维细胞的TLR 4-NF-kB依赖性促炎反应的分子机制。总之，数据表明唾液颗粒富含内毒素，但它主要是一种热不稳定级分，在生物测定中解释了趋化因子的表达。本文的在线版本（doi：10.1186/s12903-016-0229-5）包含补充材料，可供授权用户使用。
Whole saliva provokes a substantial pro-inflammatory response in gingival fibroblasts. This raises the question whether the salivary pellet, which is used for diagnostic purposes, also has a pro-inflammatory capacity and, if yes, what the underlying mechanisms at the molecular level are. We examined the ability of extensively washed salivary pellets to provoke the expression of chemokines in gingival fibroblasts by real-time polymerase chain reaction and immunoassays. Protein composition was determined with proteomic analysis. Endotoxins were analyzed by a Limulus assay and removed by affinity chromatography. The inhibitors TAK-242 and BAY11-7082 were used to determine the involvement of the TLR4 and NF-kB signaling, respectively. Western blot was performed to detect phosphorylated p65. The experiments show that salivary pellets and the corresponding washing solution contain pro-inflammatory activity without impairing cell viability. Proteomic analysis revealed proteins with a binding capacity for lipopolysaccharides, and the Limulus assay indicated the presence of endotoxin in the salivary pellets. Blocking TLR4 with TAK-242 and depletion of endotoxins both lowered the capacity of salivary pellets to increase chemokine expression and phosphorylation of p65. BAY11-7082 suppressed chemokine expression in response to the salivary pellets. Autoclaving salivary pellets also reduced their pro-inflammatory activity. The data support the molecular mechanism of a TLR4-NF-kB-dependent pro-inflammatory response of the gingival fibroblasts exposed to preparations of washed salivary pellets. Together, the data indicate that the salivary pellet is rich in endotoxin but it is mainly a heat labile fraction that accounts for the chemokine expression in the bioassay. The online version of this article (doi:10.1186/s12903-016-0229-5) contains supplementary material, which is available to authorized users.