Grant Melissa M, Scott Ann E, Matthews John B, Griffiths Helen R, Chapple Iain L C
Periodontal Research Group, School of Dentistry, Institute of Clinical Sciences and National Institute of Health Research (NIHR) Birmingham Biomedical Research Centre, University of Birmingham and Birmingham Dental Hospital, Birmingham, UK.
Unilever Oral Care, Bebington, Wirral, UK.
J Periodontal Res. 2023 Jun;58(3):634-645. doi: 10.1111/jre.13114. Epub 2023 Mar 15.
Plaque-induced gingival inflammation (gingivitis) is ubiquitous in humans. The epithelial barrier reacts to the presence of oral bacteria and induces inflammatory cascades. The objective of this study was to investigate the mechanism by which the small molecule micronutrient curcumin could decrease inflammatory response in vitro to oral bacterium heat-killed Fusobacterium nucleatum as curcumin could be a useful compound for combatting gingivitis already consumed by humans.
H400 oral epithelial cell line was pre-conditioned with curcumin and the production of cytokines was measured by enzyme-linked immunosorbent assay (ELISA) and translocation of transcription factors was used to monitor inflammatory responses. Haem oxygenase (HO-1) expression and molecules that HO-1 releases were evaluated for their potential to reduce the quantity of cytokine production. Immunofluorescence microscopy and Western blotting were used to evaluate changes in transcription factor and enzyme location.
Pre-conditioning of H400 cells with a sub-apoptotic concentration of curcumin (20 μM) attenuated secretion of Granulocyte-Macrophage - Colony-Stimulating Factor (GM-CSF) and reduced NFkB nuclear translocation. This pre-conditioning caused an increase in nuclear Nrf2; an initial drop (at 8 h) followed by an adaptive increase (at 24 h) in glutathione; and an increase in haem oxygenase (HO-1) expression. Inhibition of HO-1 by SnPPIX prevented the curcumin-induced attenuation of GM-CSF production. HO-1 catalyses the breakdown of haem to carbon monoxide, free iron and biliverdin: the HO-1/CO anti-inflammatory pathway. Elevations in carbon monoxide, achieved using carbon monoxide releasing molecule-2 (CORM2) treatment alone abrogated F. nucleatum-induced cytokine production. Biliverdin is converted to bilirubin by biliverdin reductase (BVR). This pleiotropic protein was found to increase in cell membrane expression upon curcumin treatment.
Curcumin decreased inflammatory cytokine production induced by Fusobacterium nucleatum in H400 oral epithelial cells. The mechanism of action appears to be driven by the increase of haem oxygenase and the production of carbon monoxide.
菌斑诱导的牙龈炎症（牙龈炎）在人类中普遍存在。上皮屏障对口腔细菌的存在做出反应并引发炎症级联反应。本研究的目的是探究小分子微量营养素姜黄素能够在体外降低对口腔细菌热灭活具核梭杆菌炎症反应的机制，因为姜黄素可能是一种对防治人类已摄入的牙龈炎有用的化合物。
用姜黄素预处理H400口腔上皮细胞系，通过酶联免疫吸附测定（ELISA）测量细胞因子的产生，并利用转录因子的易位来监测炎症反应。评估血红素加氧酶（HO-1）的表达以及HO-1释放的分子减少细胞因子产生量的潜力。采用免疫荧光显微镜和蛋白质免疫印迹法评估转录因子和酶位置的变化。
用亚凋亡浓度的姜黄素（20μM）预处理H400细胞可减弱粒细胞-巨噬细胞集落刺激因子（GM-CSF）的分泌并减少NFkB核易位。这种预处理导致核Nrf2增加；谷胱甘肽先下降（8小时时）随后适应性增加（24小时时）；血红素加氧酶（HO-1）表达增加。用锡原卟啉（SnPPIX）抑制HO-1可阻止姜黄素诱导的GM-CSF产生的减弱。HO-1催化血红素分解为一氧化碳、游离铁和胆绿素：即HO-1/CO抗炎途径。单独使用一氧化碳释放分子-2（CORM2）处理使一氧化碳升高可消除具核梭杆菌诱导的细胞因子产生。胆绿素通过胆绿素还原酶（BVR）转化为胆红素。发现这种多效性蛋白在姜黄素处理后细胞膜表达增加。
姜黄素可降低H400口腔上皮细胞中具核梭杆菌诱导的炎症细胞因子产生。作用机制似乎是由血红素加氧酶的增加和一氧化碳的产生所驱动。
