Circ_0066881靶向miR-144-5p/RORA轴,减轻lps诱导的人牙周韧带细胞凋亡和炎症损伤

IF 2.8 4区 医学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Innate Immunity Pub Date : 2022-05-29 DOI:10.1177/17534259221079812
Qin Li, Zhaopeng Hu, Fang Yang, Yi Peng
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引用次数: 3

摘要

环状核糖核酸(circRNAs)参与调节各种疾病,包括牙周炎。本研究的目的是分析circ_0066881在LPS诱导的牙周膜细胞(PDLC)中的生物学作用和调控机制。Circ_00066881、微小RNA-144-5p(miR-144-5p)和类视黄酸相关孤儿受体A(RORA)水平使用逆转录定量PCR(RT-qPCR)测定。通过细胞计数试剂盒-8测定法进行细胞活力检测。通过流式细胞术和胱天蛋白酶-3活性测定来评估细胞凋亡。蛋白质分析通过蛋白质印迹完成。采用ELISA法测定炎症细胞因子。通过双荧光素酶报告基因分析和RNA免疫沉淀(RIP)分析验证了靶相互作用。在牙周炎组织中circ_0066881的水平下调。circ_0066881的过表达减轻了LPS诱导的PDLC细胞活力抑制和细胞凋亡或炎症促进。Circ_00066881可以与miR-144-5p结合。circ_0066881的保护功能是通过在PDLC中吸附miR-144-5p来实现的。Circ_00066881作为miR-144-5p海绵介导RORA水平。miR-144-5p的抑制通过靶向RORA减轻LPS诱导的细胞损伤。所有这些结果表明,circ_0066881通过miR-144-5p介导的RORA上调,部分预防了LPS诱导的PDLC细胞功能障碍。
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Circ_0066881 targets miR-144-5p/RORA axis to alleviate LPS-induced apoptotic and inflammatory damages in human periodontal ligament cells
Circular RNAs (circRNAs) are involved in the regulation of various diseases, including periodontitis. The objective of this study was to analyze the biological role and regulatory mechanism of circ_0066881 in LPS-induced periodontal ligament cells (PDLCs). Circ_0066881, microRNA-144-5p (miR-144-5p) and retinoid acid-related orphan receptor A (RORA) levels were determined using reverse transcription-quantitative PCR (RT-qPCR) assay. Cell viability detection was performed by Cell Counting Kit-8 assay. Cell apoptosis was assessed through flow cytometry and caspase-3 activity assay. The protein analysis was completed via Western blot. Inflammatory cytokines were measured by ELISA. The target interaction was validated by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The level of circ_0066881 was down-regulated in periodontitis tissues. Overexpression of circ_0066881 relieved LPS-induced cell viability inhibition and apoptosis or inflammation promotion in PDLCs. Circ_0066881 could bind to miR-144-5p. The protective function of circ_0066881 was achieved by sponging miR-144-5p in PDLCs. Circ_0066881 acts as a miR-144-5p sponge to mediate the RORA level. Inhibition of miR-144-5p attenuated LPS-induced cell injury via targeting RORA. All these results demonstrated that circ_0066881 partly prevented LPS-evoked cell dysfunction in PDLCs through miR-144-5p-mediated up-regulation of RORA.
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来源期刊
Innate Immunity
Innate Immunity 生物-免疫学
CiteScore
7.20
自引率
0.00%
发文量
20
审稿时长
6-12 weeks
期刊介绍: Innate Immunity is a highly ranked, peer-reviewed scholarly journal and is the official journal of the International Endotoxin & Innate Immunity Society (IEIIS). The journal welcomes manuscripts from researchers actively working on all aspects of innate immunity including biologically active bacterial, viral, fungal, parasitic, and plant components, as well as relevant cells, their receptors, signaling pathways, and induced mediators. The aim of the Journal is to provide a single, interdisciplinary forum for the dissemination of new information on innate immunity in humans, animals, and plants to researchers. The Journal creates a vehicle for the publication of articles encompassing all areas of research, basic, applied, and clinical. The subject areas of interest include, but are not limited to, research in biochemistry, biophysics, cell biology, chemistry, clinical medicine, immunology, infectious disease, microbiology, molecular biology, and pharmacology.
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