Knockdown of NPTX1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells
10.19723/j.issn.1671-167X.2025.01.002
- VernacularTitle:敲减NPTX1促进人骨髓间充质干细胞成骨分化
- Author:
Ting SHUAI
1
;
Yanyan GUO
;
Chunping LIN
;
Xiaomei HOU
;
Chanyuan JIN
Author Information
1. 北京大学口腔医学院·口腔医院第二门诊部,国家口腔医学中心,国家口腔疾病临床医学研究中心,口腔生物材料和数字诊疗装备国家工程研究中心,口腔数字医学北京市重点实验室,北京 100081
- Publication Type:Journal Article
- Keywords:
Bone marrow mesenchymal stem cells;
NPTX1 gene;
Osteogenic differentiation
- From:
Journal of Peking University(Health Sciences)
2025;57(1):7-12
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To initially investigate the function of neuronal pentraxin 1(NPTX1)gene on osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs).Methods:hBMSCs were induced to undergo osteogenic differentiation,and then RNA was collected at different time points,namely 0,3,7,10 and 14 d.The mRNA expression levels of key genes related with osteogenic differen-tiation,including runt-related transcription factor 2(RUNX2),alkaline phosphatase(ALP),osteocalcin(OCN),and NPTX1,were detected on the basis of quantitative real-time polymerase chain reaction(qPCR)technology.In order to establish a stable NPTX1-knockdown hBMSCs cell line,NPTX1 shRNA lentivirus was constructed and used to infect hBMSCs.ALP staining,alizarin red(AR)staining,and qPCR were employed to assess the impact of NPTX1-knockdown on the osteogenic differentiation ability of hBMSCs.Results:The results showed that during the osteogenic differentiation of hBMSCs in vitro,the mRNA expression levels of osteogenic genes RUNX2,ALP and OCN significantly increased compared with 0 d,while NPTX1 expression decreased markedly(P＜0.01)as the osteogenic induction period exten-ded.At 72 h post-infection with lentivirus,the result of qPCR indicated that the knockdown efficiency of NPTX1 was over 60％.After knocking down NPTX1 in hBMSCs,RNA was extracted from both the NPTX1-knockdown group(shNPTX1 group)and the control group(shNC group)cultured in regular proliferation medium.The results of qPCR showed that the expression levels of osteogenic-related genes RUNX2 and osterix(OSX)were significantly higher in the shNPTX1 group compared with the shNC group(P＜0.01).ALP staining revealed a significantly deeper coloration in the shNPTX1 group than in the shNC group at the end of 7 d of osteogenic induction.AR staining demonstrated a marked increase in mineralized nodules in the shNPTX1 group compared with the shNC group at the end of 14 d of osteogenic induction.Conclusion:NPTX1 exerts a modulatory role in the osteogenic differentiation of hBMSCs,and its knockdown has been found to enhance the osteogenic differentiation of hBMSCs.This finding implies that NPTX1 could potentially serve as a therapeutic target for the treatment of osteogenic abnormalities,including osteoporosis.
