Actin alpha 2, smooth muscle, a transforming growth factor-β1-induced factor, regulates collagen production in human periodontal ligament cells via Smad2/3 pathway
Actin alpha 2, smooth muscle, a transforming growth factor-β1-induced factor, regulates collagen production in human periodontal ligament cells via Smad2/3 pathway
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DOI：
10.1016/j.jds.2022.08.030
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发表时间：
2023-03-21
影响因子：
3.5
通讯作者：
Wada, Naohisa
中科院分区：
文献类型：
--
作者：
Fakatava, Naati;Mitarai, Hiromi;Wada, Naohisa
背景/目的：肌动蛋白α 2，平滑肌（ACTA 2）是一种肌动蛋白亚型，形成细胞骨架。肌动蛋白在许多细胞功能中起着至关重要的作用。ACTA 2是功能性牙周膜（PDL）成纤维细胞的标志物，并被转化生长因子-β 1（TGF-β 1）上调;然而，ACTA 2在PDL组织中的潜在功能尚不清楚。我们的目的是研究ACTA 2在PDL组织和细胞中的定位和潜在的功能。材料和方法：采用半定量逆转录聚合酶链反应（RT-PCR）和定量RT-PCR测定RNA表达。免疫荧光染色和Western印迹分析测定蛋白表达。可溶性和不溶性胶原蛋白的产生分别使用Sircol胶原蛋白测定和天狼星红染色进行检查。结果：ACTA 2在人原代PDL细胞和人PDL细胞系（2-23细胞）中均有表达。TGF-81上调ACTA 2、I型胶原α 1链（COL 1A 1）、骨膜蛋白（POSTIN）和原纤维蛋白-I（FBN 1）的表达以及2-23细胞中可溶性和不溶性胶原的产生。然而，与TGF-β 1刺激的对照细胞相比，通过siRNA消耗ACTA 2强烈抑制PDL相关基因表达和胶原蛋白产生。结论：ACTA 2通过Smad 2/3磷酸化在PDL相关标志物表达和胶原合成中起重要作用。我们的研究结果可能有助于开发新的和有效的牙周治疗。(c)2022中华民国口腔医学会Elsevier B. V.的出版服务。这是一篇在CC BY-NC-ND许可证下的开放获取文章（http：//creativecommons.org/licenses/by-nc-nd/4.0/）。
Background/purpose: Actin alpha 2, smooth muscle (ACTA2) is an actin isoform that forms the cytoskeleton. Actin plays a crucial role in numerous cellular functions. ACTA2 is a marker of functional periodontal ligament (PDL) fibroblasts and is upregulated by transforming growth factor-beta 1 (TGF-beta 1); however, the underlying function of ACTA2 in PDL tissue is unknown. We aimed to examine the localization and potential function of ACTA2 in PDL tissues and cells.Materials and methods: RNA expression was determined using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and quantitative RT-PCR. Protein expression was determined using immunofluorescence staining and Western blot analysis. Soluble and insoluble collagen production was examined using the Sircol collagen assay and picrosirius red staining, respectively. Small interfering RNA (siRNA) was used for knockdown assay to examine the effect of ACTA2 in human PDL cells.Results: ACTA2 expression was observed in human primary PDL cells and PDL cell line (2-23 cells). TGF-81 upregulated ACTA2, collagen type I alpha1 chain (COL1A1), periostin (POSTN), and fibrillin-I(FBN1) expression and soluble and insoluble collagen production in 2-23 cells. However, ACTA2 depletion by siRNA strongly suppressed PDL-related gene expression and collagen production compared with those of TGF-beta 1-stimulated control cells. Furthermore, ACTA2 knockdown significantly suppressed the phosphorylation of Smad2 andConclusion: ACTA2 plays a crucial role in PDL-related marker expression and collagen production via Smad2/3 phosphorylation. Our findings might contribute to the development of novel and effective periodontal therapies. (c) 2022 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).