大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响  

Effects of Escherichia Coli Lipopolysaccharide on Biological Behaviors of Rat Dental Follicle Stem Cells

作　　者：姜震 颜燕宏 韩雪 蒋备战[1] JIANG Zhen;YAN Yanhong;HAN Xue;JIANG Beizhan(Department of Pediatrics,School and Hospital of Stomatology,Tongji University,Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Shanghai 200072,China)

机构地区：[1]同济大学口腔医学院·同济大学附属口腔医院儿童口腔科,上海牙组织修复与再生工程技术研究中心,上海200072

出　　处：《口腔医学研究》2023年第6期534-540,共7页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:82000995);上海市卫生健康委员会科研项目(编号:202040095);上海申康医院发展中心新兴前沿技术联合攻关项目(编号:SHDC12023115)。

摘　　要：目的:探究大肠杆菌脂多糖(E.coli LPS)对大鼠牙囊干细胞(rDFSCs)生物学行为的影响。方法:体外培养鉴定rDFSCs,使用不同浓度(0、1、10μg/mL)E.coli LPS处理细胞,通过qRT-PCR检测炎症因子的表达;CCK-8法检测细胞增殖情况;划痕实验和Transwell实验检测细胞迁移能力;碱性磷酸酶(ALP)染色和茜素红染色分别评估rDFSCs的矿化情况,并利用qRT-PCR检测其成骨相关基因表达水平。结果:相比于对照组,LPS组IL-6、IL-1β及TNF-α的mRNA表达水平升高(P<0.05);1μg/mL LPS促进rDFSCs增殖及迁移能力(P<0.05),而10μg/mL LPS对其增殖、迁移影响不明显;LPS处理后ALP染色降低、矿化结节数量减少,且成骨标志物RUNX2、COL1及OPN表达下降(P<0.05)。结论:E.coli LPS模拟的炎症微环境下,rDFSCs成骨分化能力受到明显抑制,低浓度(1μg/mL)LPS刺激可促进rDFSCs的增殖和迁移。Objective:To investigate the effects of Escherichia coli lipopolysaccharide(E.coli LPS)on biological behaviors of rat dental follicle stem cells(rDFSCs).Methods:rDFSCs were cultured and identified,and then were treated with different concentrations of E.coli LPS(0,1,10μg/mL).The mRNA levels of pro-inflammatory genes were measured by quantitative real-time fluorescence polymerase chain reaction(qRT-PCR).The effects of LPS on proliferation and migration ability of rDFSCs were assessed by CCK-8 and scratch test plus transwell assay,respectively.The osteogenic capability of rDFSCs was detected by alkaline phosphatase(ALP)staining and alizarin red staining,and their osteogenic-related gene mRNA levels were measured by qRT-PCR.Results:The gene expressions of IL-6,IL-1β,and TNF-αwere significantly upregulated under 1 or 10μg/mL LPS treatment(P<0.05).LPS at 1μg/mL significantly promoted the proliferation and migration of rDFSCs(P<0.05),while a higher concentration(10μg/mL)did not show any improvement.ALP staining intensity and the number of mineralized nodules decreased,and the expressions of osteogenic markers RUNX2,COL1,and OPN were inhibited in LPS-treated group(P<0.05).Conclusion:The osteogenic ability of rDFSCs was significantly inhibited under inflammatory microenvironment simulated by E.coli LPS,and LPS promoted the proliferation and migration of rDFSCs at low concentration.

关 键 词：脂多糖 牙囊干细胞 细胞增殖 细胞迁移 成骨分化 

分 类 号：R781.34[医药卫生—口腔医学]