摘要
目的:研究蛋白激酶R样内质网激酶(PERK)-转录活化因子4(ATF4)通路对增龄相关牙周膜干细胞成骨分化减弱的调控机制。方法:提取年轻和老年个体来源的牙周膜干细胞(Y-PDLSCs,O-PDLSCs),实时荧光定量PCR(qRT-PCR)和茜素红染色比较Y-PDLSCs和O-PDLSCs的成骨能力,qRT-PCR检测Y-PDLSCs与O-PDLSCs中PERK通路的葡萄糖调节蛋白78(GRP78)、CCAAT增强子结合蛋白同源蛋白(CHOP)、PERK和ATF4的表达。PERK siRNA抑制O-PDLSCs的PERK-ATF4通路,PERK过表达质粒激活Y-PDLSCs的PERK-ATF4通路,检测成骨能力。结果:O-PDLSCs的成骨能力降低,GRP78、CHOP、PERK、ATF4的表达升高(P<0.05)。抑制PERK-ATF4通路上调O-PDLSCs的ALPL、RUNX2和OCN的表达(P<0.05),促进矿化能力;激活PERK-ATF4通路降低Y-PDLSCs的成骨能力。结论:PERK-ATF4通路的上调导致O-PDLSCs成骨分化潜能降低。
Objective:To study the mechanism of protein kinase R-like ER kinase(PERK)-activating transcription factor-4(ATF4)pathway on age-related decreased osteogenic differentiation of periodontal ligament stem cells(PDLSCs).Methods:The PDLSCs from the young and the old(Y-PDLSCs,O-PDLSCs)were in vitro cultured.The osteogenic ability of PDLSCs were examined by qRT-PCR and Alizarin red staining.The PERK pathway related genes of glucose regulated protein78(GRP78),CCAAT enhancer binding protein homologous protein(CHOP),PERK and ATF4 expression in PDLSCs were detected by qRT-PCR.After inhibiting PERK-ATF4 pathway through transfecting PERK siRNA into O-PDLSCs and activating PERK-ATF4 pathway through transfecting PERK overexpression plasmid into Y-PDLSCs,the osteogenic ability of the cells was observed.Results:The osteogenic ability was decreased in O-PDLSCs.The expression of GRP78,CHOP,PERK and ATF4 in O-PDLSCs were higher than that in Y-PDLSCs(P<0.05).Inhibition of PERK-ATF4 pathway could upregulate the expression of ALPL,RUNX2 and OCN and promote the mineralization ability of O-PDLSCs,and activation of PERK-ATF4 pathway could inhibit the osteogenic ability of Y-PDLSCs.Conclusion:The activation of PERK-ATF4 pathway may lead to age-related decreased osteogenic ability of PDLSCs.
作者
张曦戈
费栋栋
王垭铮
张杨
王晓雨
葛晓彤
王勤涛
ZHANG Xige;FEI Dongdong;WANG Yazheng;ZHANG Yang;WANG Xiaoyu;GE Xiaotong;WANG Qintao(State Key Laboratory of Military Stomatology&National Clinical Research Center for Oral Diseases&Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture,Department of Periodontology,School of Stomatology,The Fourth Military Medical University,China,Xi'an 710032;Department of Stomatology,the Seventh Medical Center of PLA General Hospital;Air Force Hospital of the Eastern Theater of the Chinese People's Liberation Army)
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2022年第2期224-228,共5页
Journal of Practical Stomatology
基金
国家自然科学基金(编号:81771069)。
