Ullah Mafaz, Cox Stephen, Kelly Elizabeth, Moore Malcolm A S, Zoellner Hans
The Cellular and Molecular Pathology Research Unit, The Faculty of Dentistry, Westmead Centre for Oral Health, Westmead Hospital, The University of Sydney, Westmead, NSW, Australia.
The Department of Oral Surgery, The Faculty of Dentistry, Westmead Centre for Oral Health, Westmead Hospital, The University of Sydney, Westmead, NSW, Australia.
J Oral Pathol Med. 2015 Sep;44(8):591-601. doi: 10.1111/jop.12276. Epub 2014 Dec 19.
Areca nut chewing is associated with oral submucous fibrosis (OSF). Raised vascular basic fibroblast growth factor may induce fibrosis. Arecoline is a muscarinic alkaloid in areca nut, which we earlier reported causes injury and necrosis of human endothelium.
Human umbilical vein endothelial cells were exposed to arecoline with or without tumor necrosis factor-α, and separately to acetylcholine, muscarine, or nicotine. Protein levels of basic fibroblast growth factor, as well as the inflammatory cytokines: granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor, and Interleukins-6, 1-α and 1-β, were determined by enzyme-linked immunosorbent assay. mRNA levels were established by real-time reverse transcription polymerase chain reaction.
Basic fibroblast growth factor was released into the culture medium at arecoline levels causing necrosis (P < 0.05). This contrasted with an opposite effect of arecoline on levels of the inflammatory cytokines (P < 0.05). Tumor necrosis factor-α increased IL-6 and granulocyte-macrophage colony stimulated factor, but arecoline reduced this stimulated expression (P < 0.05). Arecoline had no effect on mRNA for basic fibroblast growth factor, although there was reduced mRNA for the separate inflammatory cytokines studied. The effect of acetylcholine, muscarine, and nicotine was minimal and dissimilar to that of arecoline.
Data raise the possibility that arecoline-induced, vascular basic fibroblast growth factor contributes to OSF, by combining increased growth factor expression with endothelial necrosis, and thus driving fibroblast proliferation.
嚼槟榔与口腔黏膜下纤维化（OSF）有关。血管中碱性成纤维细胞生长因子升高可能会诱发纤维化。槟榔碱是槟榔中的一种毒蕈碱生物碱，我们之前报道其可导致人内皮细胞损伤和坏死。
将人脐静脉内皮细胞暴露于有或无肿瘤坏死因子-α的槟榔碱中，并分别暴露于乙酰胆碱、毒蕈碱或尼古丁中。通过酶联免疫吸附测定法测定碱性成纤维细胞生长因子以及炎性细胞因子（粒细胞集落刺激因子（G-CSF）、粒细胞巨噬细胞集落刺激因子、白细胞介素-6、1-α和1-β）的蛋白水平。通过实时逆转录聚合酶链反应确定mRNA水平。
在导致坏死的槟榔碱水平下，碱性成纤维细胞生长因子释放到培养基中（P < 0.05）。这与槟榔碱对炎性细胞因子水平的相反作用形成对比（P < 0.05）。肿瘤坏死因子-α增加白细胞介素-6和粒细胞巨噬细胞集落刺激因子，但槟榔碱降低这种刺激表达（P < 0.05）。槟榔碱对碱性成纤维细胞生长因子的mRNA没有影响，尽管所研究的单独炎性细胞因子的mRNA有所减少。乙酰胆碱、毒蕈碱和尼古丁的作用最小，且与槟榔碱的作用不同。
数据表明，槟榔碱诱导的血管碱性成纤维细胞生长因子可能通过将生长因子表达增加与内皮细胞坏死相结合，从而驱动成纤维细胞增殖，进而导致口腔黏膜下纤维化。
