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脂多糖通过上调细胞周期蛋白D1、细胞周期蛋白A和细胞周期蛋白B1来增强人牙周膜细胞的增殖。

lipopolysaccharide enhances the proliferation of human periodontal ligament cells via upregulation of cyclin D1, cyclin A and cyclin B1.

作者信息

Lu Jiajing, Hu Yajing, Tang Zhongyuan, Zhang Chengfei, Jin Lijian, Gu Min, Yang Yanqi

机构信息

Division of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR 999077, P.R. China.

Department of Orthodontics, School of Medical Technology, Taizhou Polytechnic College, Taizhou, Jiangsu 225300, P.R. China.

出版信息

Exp Ther Med. 2022 Jan;23(1):2. doi: 10.3892/etm.2021.10925. Epub 2021 Oct 26.

Abstract

Human periodontal ligament cells (hPDLCs) play a notable role in periodontal tissue homeostasis and regeneration. However, the effect of lipopolysaccharide (-LPS) on the proliferation of hPDLCs remains unclear. The present study investigated the effects of -LPS on the proliferation profile of hPDLCs, and the involvement of cyclins and cyclin-dependent kinases in the process. hPDLCs were treated with -LPS, and cell proliferation and cycle were detected using Cell Counting Kit-8 assays and flow cytometry. The mRNA expression levels of the cyclins and cyclin-dependent kinases (CDKs), including cyclins A, B1, D1 and D2 and CDK1, 2 and 4, were detected using reverse transcription-quantitative PCR. The protein expression levels of cyclins A, B1 and D1 were analysed using western blotting. The proliferation of hPDLCs was significantly increased after treatment with -LPS at the concentrations of 0.001, 0.01, 0.1, 1 and 10 µg/ml for 24, 36 and 48 h compared with the cells cultured without LPS (P<0.01). The proliferation index of hPDLCs was significantly enhanced after treatment with -LPS (0.0001, 0.001, 0.01, 0.1, 1 and 10 µg/ml) for 24 h (P<0.01). However, the S-phase fraction (SPF) only significantly increased after treatment with -LPS at 0.01 µg/ml for 24 h (P<0.05), while the G/M-phase fraction increased (P<0.01) and the G/G-phase fraction decreased (P<0.01) compared with the controls. The proliferation index and SPF increased, peaked at 24 h and then decreased at 48 h in both -LPS-stimulated and control groups. Notably, -LPS significantly upregulated the expression levels of cyclins D1, A and B1 after 24 h compared with those in the controls. Overall, the present study indicated that -LPS may enhance the proliferation of hPDLCs, potentially through upregulation of cyclins D1, A and B1.

摘要

人牙周膜细胞(hPDLCs)在牙周组织稳态和再生中发挥着显著作用。然而,脂多糖(-LPS)对hPDLCs增殖的影响仍不清楚。本研究调查了-LPS对hPDLCs增殖谱的影响,以及细胞周期蛋白和细胞周期蛋白依赖性激酶在该过程中的作用。用-LPS处理hPDLCs,使用细胞计数试剂盒-8法和流式细胞术检测细胞增殖和细胞周期。使用逆转录定量PCR检测细胞周期蛋白和细胞周期蛋白依赖性激酶(CDKs)的mRNA表达水平,包括细胞周期蛋白A、B1、D1和D2以及CDK1、2和4。使用蛋白质印迹法分析细胞周期蛋白A、B1和D1的蛋白表达水平。与未用LPS培养的细胞相比,用0.001、0.01、0.1、1和10μg/ml的-LPS处理24、36和48小时后,hPDLCs的增殖显著增加(P<0.01)。用-LPS(0.0001、0.001、0.01、0.1、1和10μg/ml)处理24小时后,hPDLCs的增殖指数显著提高(P<0.01)。然而,仅在0.01μg/ml的-LPS处理24小时后,S期分数(SPF)才显著增加(P<0.05),而与对照组相比,G/M期分数增加(P<0.01),G/G期分数降低(P<0.01)。在-LPS刺激组和对照组中,增殖指数和SPF均升高,在24小时达到峰值,然后在48小时下降。值得注意的是,与对照组相比,24小时后-LPS显著上调了细胞周期蛋白D1、A和B1的表达水平。总体而言,本研究表明-LPS可能通过上调细胞周期蛋白D1、A和B1来增强hPDLCs的增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05d4/8593868/7776eecdc20f/etm-23-01-10925-g00.jpg

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