摘要
目的:研究组蛋白去乙酰化酶(HDAC5)对成牙本质细胞增殖及成牙/成骨分化的影响。方法:收集临床因治疗拔除的健康和牙髓炎状态的第三磨牙进行免疫荧光染色观察HDAC5在牙髓中的表达情况,慢病毒载体转染成牙本质细胞系(OLCs),建立过表达HDAC5的OLCs,CCK8实验检测细胞增殖,碱性磷酸酶(ALP)染色及活性检测、茜素红染色观察矿化能力,Western blot检测I型胶原(COL1α)、牙本质涎磷蛋白(DSPP)、Runt相关转录因子2(Runx2)、骨钙蛋白(OCN)、骨涎蛋白(BSP)、牙本质基质蛋白1(DMP-1)的蛋白表达。结果:与健康牙髓组织相比,炎症牙髓成牙本质细胞层见高表达的HDAC5,过表达HDAC5后OLCs增殖活力抑制,ALP活性降低,矿化结节形成减少,成牙/成骨相关蛋白表达均降低(P<0.05)。结论:HDAC5在牙髓炎成牙本质细胞层高表达且抑制OLCs的增殖和成牙/成骨向分化。
Objective:To investigate the effects of histone deacetylases5(HDAC5)on the proliferation and odontogenic/osteogenic differentiation of odontoblasts.Methods:Extracted healthy and pulpitis molars were respectively collected from clinical treatment and immunofluorescent staining was performed to detect the expression of HDAC5 in pulp.Odontoblast-lineage cells(OLCs)were transfected with lentiviral vector carrying HDAC5 gene.The cell proliferation was detected by CCK8 assay.The capacity of mineralization was examined by ALP assay and alizarin red staining.Western blot was used to measure the protein levels of collagen type 1 alpha(COL1α),dentin sialophosphoprotein(DSPP),runt-related transcription factor 2(Runx2),osteocalcin(OCN),bone sialoprotein(BSP),and dentin matrix phosphoprotein-1(DMP-1).Results:Higher expression of HDAC5 was observed in the dentinoblastic zone and pulp tissue in the pulpitis group compared with the healthy pulp tissue.In the HDAC5 gene transfection group,the proliferation of the cells was inhibited(P<0.0001).ALP activity and number of mineralized nodes was reduced(P<0.001),and accompanied by decreased expression of odonto/osteoblast related proteins(P<0.05).Conclusion:HDAC5 is highly expressed in pulp tissue of pulpitis teeth and overexpression of HDAC5 in OLCs can inhibit the proliferation and odontogenic/osteogenic differentiation of OLCs.
作者
白玉
柳鑫
程小刚
郭倩
何文喜
余擎
BAI Yu;LIU Xin;CHENG Xiaogang;GUO Qian;HE Wenxi;YU Qing(State Key Laboratory of Military Stomatology,National Clinical Research Center for Oral Diseases,Shanxi Key Laboratory of Stomatology,Department of Operative Dentistry&Endodontics,The Third Affiliated Hospital of Air Force Medical University,Xi'an,710032 China;Department of Stomatology,Air Force Medical Center,Air Force Medical University,Beijing)
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2022年第6期737-742,共6页
Journal of Practical Stomatology
基金
国家自然科学基金(编号:81970932,81771060)
西安市科技计划项目(编号:XA2020-YXYJ-0429)。
