College of Dentistry, The University of Tennessee Health Science Center, Memphis, TN, USA.
Department of Bioscience Research, The University of Tennessee Health Science Center, Memphis, TN, USA.
J Periodontal Res. 2020 Oct;55(5):762-783. doi: 10.1111/jre.12765. Epub 2020 Jun 20.
The aim of this study is to understand the role of cannabinoid type 2 receptor (CB2R) during periodontal inflammation and to identify anti-inflammatory agents for the development of drugs to treat periodontitis (PD).
Cannabinoid type 2 receptor is found in periodontal tissue at sites of inflammation/infection. Our previous study demonstrated anti-inflammatory responses in human periodontal ligament fibroblasts (hPDLFs) via CB2R ligands.
Anandamide (AEA), HU-308 (agonist), and SMM-189 (inverse agonist) were tested for effects on IL-1β-stimulated cytokines, chemokines, and angiogenic and vascular markers expressed by hPDLFs using Mesoscale Discovery V-Plex Kits. Signal transduction pathways (p-c-Jun, p-ERK, p-p-38, p-JNK, p-CREB, and p-NF-kB) were investigated using Cisbio HTRF kits. ACTOne and Tango™ -BLA functional assays were used to measure cyclic AMP (cAMP) and β-arrestin activity.
IL-1β stimulated hPDLF production of 18/39 analytes, which were downregulated by the CB2R agonist and the inverse agonist. AEA exhibited pro-inflammatory and anti-inflammatory effects. IL-1β increased phosphoproteins within the first hour except p-JNK. CB2R ligands attenuated p-p38 and p-NFĸB, but a late rise in p-38 was seen with HU-308. As p-ERK levels declined, a significant increase in p-ERK was observed later in the time course by synthetic CB2R ligands. P-JNK was significantly affected by SMM-189 only, while p-CREB was elevated significantly by CB2R ligands at 180 minutes. HU-308 affected both cAMP and β-arrestin pathway. SMM-189 only stimulated cAMP.
The findings that CB2R agonist and inverse agonist may potentially regulate inflammation suggest that development of CB2R therapeutics could improve on current treatments for PD and other oral inflammatory pathologies.
本研究旨在探讨大麻素受体 2（CB2R）在牙周炎症中的作用，并寻找抗炎药物用于治疗牙周炎（PD）。
CB2R 存在于炎症/感染部位的牙周组织中。我们之前的研究表明，CB2R 配体在人牙周膜成纤维细胞（hPDLFs）中具有抗炎反应。
使用 Mesoscale Discovery V-Plex 试剂盒检测大麻素类似物（AEA）、HU-308（激动剂）和 SMM-189（反向激动剂）对 IL-1β 刺激的 hPDLFs 表达的细胞因子、趋化因子、血管生成和血管标志物的影响。使用 Cisbio HTRF 试剂盒检测信号转导途径（p-c-Jun、p-ERK、p-p-38、p-JNK、p-CREB 和 p-NF-κB）。使用 ACTOne 和 Tango™-BLA 功能测定法测量环磷酸腺苷（cAMP）和β-arrestin 活性。
IL-1β 刺激 hPDLF 产生 18/39 种分析物，CB2R 激动剂和反向激动剂均下调这些分析物的表达。AEA 表现出促炎和抗炎作用。IL-1β 在第一个小时内增加了磷酸化蛋白，除了 p-JNK 之外。CB2R 配体减弱了 p-p38 和 p-NFκB，但 HU-308 则导致 p-38 后期增加。随着 p-ERK 水平下降，合成 CB2R 配体在时间过程中观察到 p-ERK 水平显著增加。仅 SMM-189 显著影响 p-JNK，而 CB2R 配体在 180 分钟时显著升高 p-CREB。HU-308 影响 cAMP 和 β-arrestin 途径。SMM-189 仅刺激 cAMP。
CB2R 激动剂和反向激动剂可能潜在地调节炎症的发现表明，CB2R 治疗药物的开发可以改善 PD 和其他口腔炎症性疾病的现有治疗方法。
