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Low-power laser irradiation inhibits arecoline-induced fibrosis: an in vitro study

机译:低功率激光辐照抑制槟榔碱诱导的纤维化:一项体外研究

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摘要

Oral submucous fibrosis (OSF) is a potentially malignant disorder that is characterized by a progressive fibrosis in the oral submucosa. Arecoline, an alkaloid compound of the areca nut, is reported to be a major aetiological factor in the development of OSF. Low-power laser irradiation (LPLI) has been reported to be beneficial in fibrosis prevention in different damaged organs. The aim of this study was to investigate the potential therapeutic effects of LPLI on arecoline-induced fibrosis. Arecoline-stimulated human gingival fibroblasts (HGFs) were treated with or without LPLI. The expression levels of the fibrotic marker genes alpha-smooth muscle actin (α-SMA) and connective tissue growth factor (CTGF/CCN2) were analysed by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) and western blots. In addition, the transcriptional activity of CCN2 was further determined by a reporter assay. The results indicated that arecoline increased the messenger RNA and protein expression of CCN2 and α-SMA in HGF. Interestingly, both LPLI and forskolin, an adenylyl cyclase activator, reduced the expression of arecoline-mediated fibrotic marker genes and inhibited the transcriptional activity of CCN2. Moreover, pretreatment with SQ22536, an adenylyl cyclase inhibitor, blocked LPLI's inhibition of the expression of arecoline-mediated fibrotic marker genes. Our data suggest that LPLI may inhibit the expression of arecoline-mediated fibrotic marker genes via the cAMP signalling pathway.
机译:口腔粘膜下纤维化(OSF)是一种潜在的恶性疾病,其特征是口腔粘膜下层进行性纤维化。槟榔碱是槟榔的生物碱化合物,据报道是OSF发生的主要病因。据报道,低功率激光辐照(LPLI)在预防不同受损器官的纤维化方面是有益的。这项研究的目的是调查LPLI对槟榔碱诱导的纤维化的潜在治疗作用。槟榔碱刺激的人牙龈成纤维细胞(HGF)用或不用LPLI治疗。通过定量实时逆转录聚合酶链反应(RT-PCR)和western印迹分析了纤维化标记基因α-平滑肌肌动蛋白(α-SMA)和结缔组织生长因子(CTGF / CCN2)的表达水平。另外,CCN2的转录活性通过报告基因测定法进一步确定。结果表明槟榔碱可增加HGF中CCN2和α-SMA的信使RNA和蛋白表达。有趣的是,LPLI和Forskolin(一种腺苷酸环化酶激活剂)均降低了槟榔碱介导的纤维化标记基因的表达,并抑制了CCN2的转录活性。此外,用腺苷酸环化酶抑制剂SQ22536预处理可阻断LPLI对槟榔碱介导的纤维化标记基因表达的抑制。我们的数据表明,LPLI可能通过cAMP信号通路抑制槟榔碱介导的纤维化标记基因的表达。

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