巨噬细胞炎性蛋白1α对人牙周膜干细胞生物学行为的影响  

Effect of macrophage inflammatory protein-1 alpha on the biological behavior of human periodontal ligament stem cells

作　　者：王钊鑫 尼加提•吐尔逊 代慧娟 王俊祥 霞黒达•依拉尔江 李淑慧[1] Wang Zhaoxin;Nijati·Tursun;Dai Huijuan;Wang Junxiang;Xiaheida·Yilaljiang;Li Shuhui(Department of Stomatology,Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830063,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第二附属医院口腔科,新疆维吾尔自治区乌鲁木齐市830063

出　　处：《中国组织工程研究》2023年第10期1521-1527,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区科学基金(2019D01C231),项目负责人:李淑慧。

摘　　要：背景:目前大量研究集中于炎症因子对人牙周膜干细胞的影响,然而关于趋化因子巨噬细胞炎性蛋白1α(macrophage inflammatory protein-1α,MIP-1α)在人牙周膜干细胞中的表达,国内外相关文献报道较少。目的:探究不同质量浓度MIP-1α对人牙周膜干细胞增殖、凋亡及迁移的影响及与Notch信号通路的相关性。方法:将12-25岁患者(均签署知情同意书)拔下的健康正畸减数前磨牙,通过组织块法联合酶消化法分离培养人牙周膜干细胞。将第3代对数生长期人牙周膜干细胞分为对照组和1,10,100 mg/L MIP-1α组,干预第1,3,5,7天采用CCK-8法检测细胞增殖能力,干预48 h采用流式检测细胞凋亡能力,Transwell小室检测细胞迁移能力,ELISA检测分泌肿瘤坏死因子α水平,qRT-PCR、Western blot检测Notch1、Hes1mRNA和蛋白表达量。结果与结论:(1)1 mg/L MIP-1α对人牙周膜干细胞生物学行为无明显影响,10 mg/L MIP-1α显著促进人牙周膜干细胞的增殖及迁移,而100 mg/L MIP-1α显著促进人牙周膜干细胞的凋亡,抑制人牙周膜干细胞的增殖和迁移;(2)1 mg/L MIP-1α对人牙周膜干细胞中Notch1及Hes1蛋白表达无明显影响;10 mg/L MIP-1α对人牙周膜干细胞中Notch1蛋白表达无显著影响,但可下调细胞中Hes1蛋白表达水平,100 mg/L MIP-1α可上调细胞中Notch1及Hes1蛋白表达水平;(3)1 mg/L MIP-1α组人牙周膜干细胞分泌的肿瘤坏死因子α水平与对照组无显著差异;10 mg/L MIP-1α促进肿瘤坏死因子α分泌;100 mg/L MIP-1α抑制肿瘤坏死因子α分泌;(4)结果表明:10 mg/L MIP-1α促进人牙周膜干细胞增殖、迁移及肿瘤坏死因子α分泌,其机制可能与下调Notch信号通路相关;100 mg/L MIP-1α抑制人牙周膜干细胞增殖、迁移及肿瘤坏死因子α分泌,促进人牙周膜干细胞凋亡,其机制可能与上调Notch信号通路有关。BACKGROUND:A large number of studies focus on the effect of inflammatory factors on human periodontal ligament stem cells;however,there are few reports on the expression of chemokine macrophage inflammatory protein-1α in human periodontal ligament stem cells at home and abroad.OBJECTIVE:To investigate the effects of different mass concentrations of macrophage inflammatory protein 1α on the proliferation,apoptosis,and migration of human periodontal ligament stem cells and the correlation of Notch signaling pathway.METHODS:Healthy orthodontic reduced premolars were extracted from patients aged 12-25 years(all signed informed consent),and human periodontal ligament stem cells were isolated and cultured by tissue block method combined with enzymatic digestion.Passage 3 human periodontal ligament stem cells at logarithmic growth stage were divided into control group,1,10,100 mg/L macrophage inflammatory protein-1α groups.Cell proliferation was detected by CCK-8 assay at 1,3,5,and 7 days.At 48 hours,the apoptotic ability of each group was detected by flow cytometry.Cell migration ability of each group was detected by Transwell chamber test.ELISA was used to detect the level of secreted tumor necrosis factor α.qRT-PCR and western blot assay were used to detect the mRNA and protein expression levels of Notch1 and Hes1.RESULTS AND CONCLUSION:(1) 1 mg/L macrophage inflammatory protein-1α had no significant effect on the biological behavior of human periodontal ligament stem cells.10 mg/L macrophage inflammatory protein-1α significantly promoted the proliferation and migration of human periodontal ligament stem cells,and 100 mg/L macrophage inflammatory protein-1α significantly promoted the apoptosis of human periodontal ligament stem cells,inhibited the proliferation and migration of human periodontal ligament stem cells.(2) 1 mg/L macrophage inflammatory protein-1α had no significant effect on the expression of Notch1 and Hes1 proteins in human periodontal ligament stem cells.10 mg/L macrophage inflammatory protei

关 键 词：人牙周膜干细胞 巨噬细胞炎性蛋白1Α NOTCH信号通路 增殖 迁移 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学] R542.2