长链非编码RNA MALAT1对槟榔碱诱导的颊黏膜成纤维细胞活化能力影响的研究

doi:10.13701/j.cnki.kqyxyj.2024.07.010

摘要/Abstract

摘要： 目的:本研究旨在明确长链非编码RNA(long non-coding RNA,lncRNA) MALAT1对槟榔碱诱导的颊黏膜成纤维细胞(buccal mucosal fibroblasts, BMFs)活化能力影响的作用,为寻找新的口腔黏膜下纤维化(oral submucous fibrosis,OSF)治疗途径提供理论基础。方法:体外培养BMFs,采用不同浓度槟榔碱刺激BMFs,在细胞水平上构建OSF疾病模型。采用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞活力;Transwell法和胶原凝胶收缩法检测BMFs活化;实时荧光定量聚合酶链反应(quantitative real-time PCR, qRT-PCR)检测BMFs中平滑肌肌动蛋白-α(α-smooth muscle actin, α-SMA)和lncRNA MALAT1的表达情况;siRNA-MALAT1瞬时转染到BMFs中,评估槟榔碱刺激下BMFs迁移和收缩能力的变化。结果:10 μg/mL槟榔碱是体外构建OSF疾病模型的最佳浓度,此时细胞内α-SMA和lncRNA MALAT1表达升高。Transwell实验和胶原凝胶收缩实验提示敲低lncRNA MALAT能抑制槟榔碱诱导的BMFs收缩和迁移。结论:lncRNA MALAT1对槟榔碱诱导的BMFs激活,细胞收缩和迁移有重要作用。

关键词: 长链非编码RNA MALAT1, 口腔黏膜下纤维化, 槟榔碱, 成纤维细胞

Abstract: Objective: To clarify the effect of long non-coding RNA MALAT1 on the activation ability of buccal mucosal fibroblasts (BMFs) induced by arecoline, and to establish theoretical basis for a new therapeutic pathway to oral submucous fibrosis (OSF). Methods: BMFs were cultured in vitro, and were stimulated with different concentrations of arecoline to construct OSF-disease models at the cellular level. CCK-8 method was used to detect cell viability. BMFs activation was detected by Transwell method and collagen gel shrinkage method. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of α-smooth muscle actin antibody (α-SMA) and lncRNA MALAT1 in BMFs. siRNA-MALAT1 was transiently transfected into BMFs to assess changes in the migration and contraction capacity of BMFs stimulated by arecoline. Results: 10 μg/mL of arecoline was the optimal concentration for constructing OSF-disease models in vitro, when intracellular expression of α-SMA and lncRNA MALAT1 was elevated. Transwell experiments and collagen gel contraction experiments suggested that knockdown lncRNA MALAT inhibited arecoline-induced BMFs contraction and migration. Conclusion: lncRNA MALAT1 plays an important role in arecoline-induced BMFs activation, cell contraction, and migration.

Key words: lncRNA MALAT1, oral submucous fibrosis, arecoline, fibroblasts

李熠洁, 邵溢朵, 蔡芸舟, 吴光栋, 孙天翱, 王月红. 长链非编码RNA MALAT1对槟榔碱诱导的颊黏膜成纤维细胞活化能力影响的研究[J]. 口腔医学研究, 2024, 40(7): 622-628.

LI Yijie, SHAO Yiduo, CAI Yunzhou, WU Guangdong, SUN Tianao, WANG Yuehong. LncRNA MALAT1 Inhibits Buccal Mucosal Fibroblasts Activities in Oral Submucous Fibrosis[J]. Journal of Oral Science Research, 2024, 40(7): 622-628.

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