昆明医科大学学报2025，Vol.46Issue(12)：30-39,10.DOI:10.12259/j.issn.2095-610X.S20251204
HDAC3靶向调控GATA-2影响牙髓干细胞成骨分化的机制
Mechanism of HDAC3-targeted Regulation of GATA-2 Affecting Osteogenic Differentiation of Dental Pulp Stem Cells
摘要
Abstract
Objective To investigate the mechanism of the HDAC3/GATA-2 molecular axis in osteogenic differentiation of human dental pulp stem cells(hDPSCs).Methods The hDPSCs cell line was cultured in osteogenic induction medium(OM).Plasmids of oe-NC,oe-HDAC3 and oe-GATA-2 were constructed and transfected into the cells.Transfection efficiency was assessed by RT-qPCR and Western blot.Alkaline phosphatase(ALP)staining and alizarin red S(ARS)staining were used to evaluate ALP activity and mineralization nodules in cells.RT-qPCR and Western Blot were used to detect the expression levels of osteogenic key indicators COL1A1,ALP,BMP2,and RUNX2.GST-pull down and Co-Immunoprecipitation(CO-IP)were used to validate the interaction of HDAC3 and GATA-2 interaction,and immunofluorescence(IF)staining was performed to detect cellular expression.Results HDAC3 expression was significantly reduced during osteogenic differentiation of hDPSCs(P<0.000 1).Overexpression of HDAC3 significantly attenuated ALP staining and activity(P=0.000 1),a s well as reduced the production of mineralized nodules in hDPSCs cells.The expression levels of COL1A1(P=0.002 9),ALP(P=0.000 1),BMP2(P=0.000 1)and RUNX2(P=0.000 7)were inhibited after overexpression of HDAC3.HDAC3 interacted with GATA-2,and overexpression of HDAC3 decreased the expression of GATA-2 in cells(P=0.002 8).Co-transfection of oe-GATA-2 reversed the inhibitory effects of HDAC3 overexpression,increasing ALP staining and activity(P=0.004 3)and mineralization nodules in cells.In the oe-HDAC3+oe-GATA-2 group,the expression levels of COL1A1(P=0.000 1),ALP(P=0.042 3),BMP2(P<0.000 1),and RUNX2(P=0.000 5)were further elevated.Conclusion HDAC3 inhibited the osteogenic differentiation of hDPSCs by targeting and negatively regulating the expression of GATA-2.关键词
人牙髓干细胞/成骨分化/HDAC3/GATA-2Key words
Human dental pulp stem cells/Osteogenic differentiation/HDAC3/GATA-2分类
医药卫生引用本文复制引用
OUYANG Qian,HUANG Yanfei,YIN Lingyun,ZHANG Lingpeng,LIANG Jing,YU Hongbin..HDAC3靶向调控GATA-2影响牙髓干细胞成骨分化的机制[J].昆明医科大学学报,2025,46(12):30-39,10.基金项目
云南省科技厅科技计划项目(202401AY070001-300) （202401AY070001-300）
云南省教育厅科学研究基金(2024J0282,2024J0279) （2024J0282,2024J0279）
昆明市卫生健康委员会科研基金(2023-08-04-001) （2023-08-04-001）
