摘要
目的探讨根吸收不同时期乳牙牙周膜干细胞(PDLSCs)通过自然杀伤细胞(NK)发挥破骨分化的调节作用。方法酶消化法和有限稀释法分离培养根吸收不同时期乳牙和恒牙与PDLSCs与NK细胞、RAW264.7细胞进行共培养。采用实时定量PCR和ELISA,检测各组共培养体系中NK细胞IFN-γ表达及其差异,以及阻断转化生长因子-β(TGF-β)后IFN-γ的表达及差异。采用实时定量PCR和Western blot检测NK细胞、RAW264.7细胞共培养体系中RAW264.7细胞核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)表达。结果根吸收中期组(B)NK细胞干扰素γ(IFN-γ)表达明显下降(P<0.05)。阻断TGF-β后,IFN-γ表达明显升高(P<0.05)。外源性加入NK细胞与PDLSCs共培养上清液后,RAW264.7细胞RANKL表达降低,OPG表达升高,RANKL/OPG比值降低。结论在乳牙生理性根吸收过程中,PDLSCs通过分泌TGF-β等细胞因子对NK细胞发挥免疫抑制作用,影响破骨分化进程。
Objective To explore the role of periodontal ligament stem cells(PDLSCs)of deciduous teeth at different stages of root absorption through natural killer cells(NK)in regulating osteoclast differentiation.Methods Enzyme digestion method and limited dilution method were used to separate PDLSCs and NK cells from permanent teeth and deciduous teeth at different stages of root absorption,and co-culture them with RAW264.7 cells.Real-time quantitative PCR and ELISA were used to detect the expression and difference of IFN-γof NK cells in the co-culture system of each group,and those of IFN-γafter blocking TGF-β.NK cells were detected by real-time quantitative PCR and Western blot.The expression of RANKL and OPG in RAW264.7 cell co-culture system was determined.Results The expression of IFN-γof NK cells in the mid-root absorption group(B)decreased significantly(P<0.05).After blocking TGF-β,the expression of IFN-γwas significantly increased(P<0.05).After exogenous addition of NK cells and PDLSCs co-culture supernatant,RANKL expression of RAW264.7 cells decreased;OPG expression increased;RANKL/OPG ratio decreased.Conclusion During the physiological root absorption process of deciduous teeth,DPDLSCs secrete TGF-βand other cytokines to exert an immunosuppressive effect on NK cells and affect the process of osteoclast differentiation.
作者
赵辛
杨宽
王子瑞
韩欣欣
汪璐璐
王小竞
ZHAO Xin;YANG Kuan;WANG Zirui;HAN Xinxin;WANG Lulu;WANG Xiaojing(State Key Laboratory of Military Stomatology,National Clinical Reaserch Center for Oral Diseases,Shaanxi Clinical Research Center for Oral Diseases,Department of Pediatric Dentistry,School of Stomatology,Fourth Military Medical University,Xi'an 710032,China)
出处
《口腔医学》
CAS
2020年第6期517-520,575,共5页
Stomatology
基金
国家自然科学基金(81670988,81800924)
陕西省自然科学基金(2019JQ-699)。
关键词
生理性根吸收
乳牙牙周膜干细胞
NK细胞
physiological root absorption
periodontal ligament stem cells of deciduous teeth
NK cells