虾青素对槟榔碱诱导的口腔上皮损伤的保护效应及机制初探

Research on the Protective Effect and Mechanism of Astaxanthin on Arecoline Induced Oral Epithelial Injury

摘要:

探索虾青素（AST）对槟榔碱（ARC）诱导的口腔上皮损伤的保护效应及其作用机制。建立ARC诱导的大鼠口腔黏膜损伤模型，观察大鼠口腔黏膜组织病理学形态及人早期生长反应因子1（EGR1）的变化。测定ARC诱导的人正常口腔上皮角质（HOK）细胞中紧密连接相关蛋白ZO-1和Occludin、线粒体形态、线粒体膜电位（MMP）、线粒体活性氧（mtROS）和EGR1的变化。通过分子对接预测AST与EGR1的结合情况。大鼠口腔黏膜组织病理切片显示，与模型组相比，AST干预组中口腔黏膜上皮变薄、钉突变短症状有所改善，黏膜下相对胶原面积及Collagen Ⅰ 面积显著降低9.84%、23.01%。细胞实验结果显示，AST改善了ARC诱导的HOK细胞ZO-1和Occludin蛋白定位异常、线粒体形态损伤、MMP下降及mtROS生成过多。此外，AST抑制ARC诱导的EGR1高表达，细胞水平上能显著下调24.53%。分子对接结果显示AST与EGR1的对接结合能为－6.8697 kcal/mol，存在两个氢键作用。综上所述，AST可能通过调控EGR1和线粒体功能从而改善ARC诱导的口腔黏膜损伤，有望开发成为保护口腔黏膜的功能性食品。

Abstract:

To study the protective effect and mechanism of astaxanthin (AST) on oral epithelial injury, an arecoline (ARC) induced oral mucosal injury rat model was established. The pathological morphology, the accumulation of collagen fibers, and changes in human early growth response factor 1 (EGR1) in rat oral mucosal tissue was observed. The changes in tight junction related proteins ZO-1 and Occludin, mitochondrial morphology, mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (mtROS), and EGR1 in human oral keratinocyte (HOK) cells induced by ARC, were measured. And the binding of AST and EGR1 was predicted through molecular docking. The results on pathological of oral mucosal tissues showed that compared with the model group, the symptoms of thinning oral mucosal epithelium and short epithelial nail was improved in the AST intervention group. The relative collagen area and Collagen I area under the mucosa were significantly reduced by 9.84% and 23.01%, respectively. In vitro experiments, the results showed that AST improved the abnormal localization of ZO-1 and Occludin proteins in ARC-induced HOK cells; the abnormal mitochondrial morphological, the decreased level of MMP, and the excessive generation of mtROS were also improved. In addition, AST inhibited the expression level of EGR1 in ARC-induced HOK cells, and significantly downregulating 24.53% at the cellular level. The molecular docking results showed that the docking binding energy between AST and EGR1 was -6.8697 kcal/mol, and had two hydrogen bonding interactions. In summary, AST may improve ARC induced oral mucosal injury by regulating EGR1 and mitochondrial function, and is expected to be developed as a functional food for protecting oral mucosa.