高迁移率族蛋白B1对人牙周膜干细胞增殖、分化影响的体外研究
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摘要
目的:探讨高迁移率族蛋白B1(HMGB1)对于人牙周膜干细胞(hPDLSCs)增殖、分化的影响。方法:选取年轻患者拔除的健康第三磨牙,采用胶原酶消化组织块培养得到牙周膜细胞,并利用有限稀释法筛选获得纯化的hPDLSCs。分别以浓度为0、100、200ng/ml的HMGB1作用于hPDLSCs,于第3、5天采用MTT法检测细胞数目增殖情况;于第7天采用RT-PCR法检测细胞因子:增殖细胞核抗原(PCNA)、白细胞介素-1b(IL-1b)、肿瘤坏死因子α(TNFα)和抗酒石酸酸性磷酸酶(TRAP)的基因表达。结果:HMGB1在100、200ng/ml浓度时,在第3天和第5天hPDLSCs的OD值有明显升高,在第7天PCNA、IL-1 b、TNFα和TRAP的基因表达明显升高。结论:HMGB1对hPDLSCs的增殖具有促进作用,同时使其破骨功能加强,与牙周炎的发展有关。
Objective:To explore the effect of High mobility group box 1(HMGB1)to human periodontal ligament stem cells(hPDLSCs)on proliferation and differentiation.Methods:Periodontal ligament tissues obtained from the third molar teeth extracted from yong patient were digested with collagenase,then the periodontal ligament stem cells were isolated by limited dilution method.MTT analysis was conducted to hPDLSCs in inducing medium containing 0,100,200 ng/ml HMGB1.The differentiation of hPDLSCs cultured in medium supplemented with or without HMGB1 were measured by RT-PCR analysis on genes including proliferating cell nuclear antigen(PCNA),interleukin-1 b(IL-1 b)and tumor necrosis factor-α(TNFα)and tartrate-resistant acid phosphatase(TRAP)on day 7.Results:The hPDLSCs treated with HMGB1(100 and 200 ng/ml)proliferated faster than the cells cultured in medium on day 3 and 5.And the genes expression including PCNA,IL-1 b,TNFα and TRAP was significantly enhanced by HMGB1 in the concentration of 100 and 200 ng/ml.Conclusion:The use of proper concentrations of HMGB1 enhanced the proliferation and osteoclastic differentiation abilities of hPDLSCs,which related to the development of periodontitis.
Objective:To explore the effect of High mobility group box 1(HMGB1)to human periodontal ligament stem cells(hPDLSCs)on proliferation and differentiation.Methods:Periodontal ligament tissues obtained from the third molar teeth extracted from yong patient were digested with collagenase,then the periodontal ligament stem cells were isolated by limited dilution method.MTT analysis was conducted to hPDLSCs in inducing medium containing 0,100,200 ng/ml HMGB1.The differentiation of hPDLSCs cultured in medium supplemented with or without HMGB1 were measured by RT-PCR analysis on genes including proliferating cell nuclear antigen(PCNA),interleukin-1 b(IL-1 b)and tumor necrosis factor-α(TNFα)and tartrate-resistant acid phosphatase(TRAP)on day 7.Results:The hPDLSCs treated with HMGB1(100 and 200 ng/ml)proliferated faster than the cells cultured in medium on day 3 and 5.And the genes expression including PCNA,IL-1 b,TNFα and TRAP was significantly enhanced by HMGB1 in the concentration of 100 and 200 ng/ml.Conclusion:The use of proper concentrations of HMGB1 enhanced the proliferation and osteoclastic differentiation abilities of hPDLSCs,which related to the development of periodontitis.
引文
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[2] Slack JM.Stem cells in epithelial tissues[J].Science,2000,287(5457):1431-1433.
[3] Weissman IL.Stem cells:units of development,units of regeneration,and units in evolution[J].Cell,2000,100(1):157-168.
[4] Wang H,Bloom O,Zhang M,et al.HMG-1 as a late mediator of endotoxin lethality in mice[J].Science,1999,285(5425):248.
[5] 袁玉亮.HMGB1在AECOPD、肺炎、肺癌患者中的表达水平及临床意义研究[J].国际检验医学杂志,2017,38(7):988-990.
[6] 黄鹤,田昭涛,黎檀实.脓毒血症中固有免疫细胞的调节机制研究进展[J].中国免疫学杂志,2016,36(4):576-583.
[7] Paknejad M,Sattari M,et al.Relationships between high-mobility group protein B1 and triggering receptor expressed on myeloid cells concentrations in gingival crevicular fluid and chronic periodontitis[J].Iran J Allergy Asthma Immunol,2016,15(5):381-385.
[8] 孙钦峰,徐岩,等.高迁移率族蛋白B1对牙周膜成纤维细胞表达细胞因子影响的研究[J].华西口腔医学杂志,2010,29(4):443.
[9] Seo BM,Miura M,Gronthos S,et al.Investigation of multipotent postanal stem cells from periodontal ligament[J].Lancet,2004,364(9429):149-155.
[10] Cohen-Kaplan V,Jrbashyan J,Yanir Y,et al.Herparnase induces signal transducer and sctivator of transcription (STAT) protein phosphorylation:preclinical and clinical significance in head and neck cancer[J].J Biol Chem,2012,287(9):6668-6678.
[11] Lastic al,Rodi M,Mouiaki A.Effect of dendritic cell state and antigen-Presentation condition on resulting T-cell phenotypes and TH cytokine profiles[J].Immuunobiology,2016,221(8):862.
[12] Tan J,Zhou L,Xue P,et al.Tumor necrosis factor-alpha attenuates the osteogenic differentiation capacity of perio-dontal ligament stem cells by activating protein kinase like endoplasmic reticulum kinase signaling[J].J Periodontol,2016,87(8):1-22.
[13] Zhang Meng-meng.Bone metabolism indexes in bone remodeling[J].Chin J Osteoporos,2013,19(8):866-873.