circRASA2靶向miR-543/TRAF6轴对LPS诱导的牙周膜细胞增殖和成骨分化的影响

doi:10.19439/j.sjos.2023.02.007

摘要/Abstract

摘要： 目的: 探讨circRASA2在牙周炎中的可能作用及其潜在调控机制。方法: 采用脂多糖（LPS）诱导牙周膜细胞（PDLCs）,建立牙周炎细胞模型。CCK-8实验检测细胞增殖活性,Transwell小室实验检测细胞迁移能力,Western印迹法检测细胞中成骨分化相关蛋白表达。分别利用数据库circinteractome和starBase预测circRASA2的靶miRNA及下游靶基因,通过双荧光素酶报告基因实验验证目的基因之间的靶向关系。采用GraphPad Prism 8.0软件包对数据进行统计分析。结果: circRASA2在LPS处理的PDLCs细胞中高表达;LPS诱导PDLCs细胞增殖活力、迁移能力和成骨分化能力降低,而敲低circRASA2能促进LPS处理下PDLCs的增殖、迁移和成骨分化能力。circRASA2靶向并负调控miR-543的表达,过表达miR-543能促进LPS处理下PDLCs的增殖、迁移和成骨分化能力。TRAF6是miR-543的下游靶基因,敲低circRASA2可通过miR-543的海绵作用,下调TRAF6的表达。过表达TRAF6能逆转circRASA2敲低对PDLCs增殖、迁移和成骨分化能力的促进作用。结论: circRASA2通过miR-543/TRAF6轴在体外加速牙周炎的病理进程,通过靶向降低circRASA2的表达,可能改善牙周炎症状。

关键词: 牙周炎, 牙周膜细胞, circRASA2, miR-543, TRAF6

Abstract: PURPOSE: To investigate the possible role of circRASA2 in periodontitis and its potential regulatory mechanism. METHODS: Periodontitis cell model was established by lipopolysaccharide(LPS)-induced periodontal ligament cells(PDLCs). Cell proliferation activity was detected by CCK-8 assay, cell migration ability was detected by Transwell chamber assay, and the expression of osteogenic differentiation-related proteins in cells was detected by Western blot. The target miRNA of circRASA2 and its downstream target genes were predicted using the databases circinteractome and starBase, respectively, and the targeting relationship between the target genes was verified by dual-luciferase reporter gene experiment. GraphPad Prism 8.0 software package was used to analyze the data. RESULTS: circRASA2 was highly expressed in LPS-treated PDLCs cells. LPS-induced PDLCs cell proliferation activity, migration ability and osteogenic differentiation ability decreased, while knockdown of circRASA2 promoted proliferation, migration and osteogenic differentiation ability of PDLCs under LPS treatment. circRASA2 targeted and negatively regulated the expression of miR-543, and overexpression of miR-543 promoted proliferation, migration and osteogenic differentiation of PDLCs under LPS treatment. TRAF6 was a downstream target gene of miR-543, knockdown of circRASA2 down-regulated the expression of TRAF6 through the sponge action of miR-543. Overexpression of TRAF6 reversed the promotion of circRASA2 knockdown on proliferation, migration and osteogenic differentiation of PDLCs. CONCLUSIONS: circRASA2 accelerated the pathological process of periodontitis in vitro through miR-543/TRAF6 axis, and might improve periodontitis by targeting down the expression of circRASA2.

Key words: Periodontitis, Periodontal ligament cells, circRASA2, miR-543, TRAF6

中图分类号:

R781.4

谢冰, 杨振宇, 汤旭娜. circRASA2靶向miR-543/TRAF6轴对LPS诱导的牙周膜细胞增殖和成骨分化的影响[J]. 上海口腔医学, 2023, 32(2): 147-153.

XIE Bing, YANG Zhen-yu, TANG Xu-na. Effect of circRASA2 targeting miR-543/TRAF6 axis on LPS-induced periodontal ligament cell proliferation and osteogenic differentiation[J]. Shanghai Journal of Stomatology, 2023, 32(2): 147-153.

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