Department of Oral Biology, The Maurice and Gabriela Goldschleger School of Dental Medicine, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.
Department of Oral Biology, The Maurice and Gabriela Goldschleger School of Dental Medicine, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; Department of Periodontology and Dental Implantology, The Maurice and Gabriela Goldschleger School of Dental Medicine, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.
Arch Oral Biol. 2020 Aug;116:104766. doi: 10.1016/j.archoralbio.2020.104766. Epub 2020 May 20.
Diabetes increases the incidence/severity of periodontal diseases by inducing a chronic inflammation, driven by accumulation of AGEs (advanced glycation end products). We tested whether glycated human serum albumin (G-HSA, a form of AGE), representing a diabetic state, augments the pro-inflammatory response of human gingival fibroblasts (hGFs) to a bacterial challenge (Porphyromonas gingivalis Lipopolysaccharide (LPS)).
Primary hGFs were incubated with LPS (0.5-5 μg/mL) and G-HSA (50-200 μg/mL) and the production and gene expression of IL-1β, IL-6, IL-8, MMP-1, MCP-1, and TNFα were analyzed by Magnetic Luminex Assay and real-time PCR, respectively. Non-glycated serum albumin (HSA) served as negative control. Cytotoxicity of the 2 agents was tested with an XTT assay. NFκB activation (p65 phosphorylation) was measured with an ELISA.
P. gingivalis LPS and G-HSA were not toxic to hGFs and increased the amount of MMP-1, MCP-1, IL-6, and IL-8, (but not TNFα and IL-1β) secreted into the medium at 24 h. Control HSA had no effect. Many LPS/G-HSA combinations displayed a synergistic stimulation of these molecules. Both agents increased mRNA levels of these 4 molecules at 6 h, 12 h or both (IL-6). NFκB activation at 6 h was caused by both agents with a possible synergism at the higher concentrations.
glycated albumin augments the pro-inflammatory response of human gingival fibroblasts to P. gingivalis LPS. Thus, AGE accumulation in diabetes could aggravate periodontal inflammation by augmenting the pro-inflammatory response of host GFs to P. gingivalis, a well-recognized periopathogenic bacteria.
糖尿病通过诱导由 AGEs（糖基化终产物）积累驱动的慢性炎症，增加牙周病的发病率/严重程度。我们测试了糖化人血清白蛋白（G-HSA，AGE 的一种形式）是否代表糖尿病状态，会增强人牙龈成纤维细胞（hGF）对细菌挑战（牙龈卟啉单胞菌脂多糖（LPS））的促炎反应。
将 hGF 与 LPS（0.5-5 μg/mL）和 G-HSA（50-200 μg/mL）孵育，并通过磁珠 Luminex 测定法和实时 PCR 分别分析 IL-1β、IL-6、IL-8、MMP-1、MCP-1 和 TNFα 的产生和基因表达。非糖化血清白蛋白（HSA）作为阴性对照。用 XTT 测定法测试 2 种试剂的细胞毒性。用 ELISA 测量 NFκB 激活（p65 磷酸化）。
牙龈卟啉单胞菌 LPS 和 G-HSA 对 hGF 没有毒性，并在 24 小时增加了进入培养基的 MMP-1、MCP-1、IL-6 和 IL-8 的量（但不增加 TNFα 和 IL-1β）。对照 HSA 没有作用。许多 LPS/G-HSA 组合对这些分子表现出协同刺激作用。两种试剂均在 6 小时和 12 小时（IL-6）增加了这些 4 种分子的 mRNA 水平。两种试剂均在 6 小时引起 NFκB 激活，在较高浓度下可能存在协同作用。
糖化白蛋白增强了人牙龈成纤维细胞对牙龈卟啉单胞菌 LPS 的促炎反应。因此，糖尿病中 AGE 的积累可能通过增强宿主 GFs 对牙周病病原菌牙龈卟啉单胞菌的促炎反应来加重牙周炎症。
