沉默PGC-1α表达对人根尖牙乳头干细胞定向分化的影响

口腔生物医学 ›› 2023, Vol. 14 ›› Issue (3): 146-150.

沉默PGC-1α表达对人根尖牙乳头干细胞定向分化的影响

肖雅¹,陈路遥²,顾婷杰¹,方玉心²,李泽汉³,于金华⁴,⁴

1. 南京医科大学
2. 无
3. 南京医科大学附属口腔医院牙体牙髓科
4. 南京医科大学附属口腔医学院

收稿日期:2023-02-09 修回日期:2023-05-15 出版日期:2023-09-25 发布日期:2023-09-21
通讯作者: 于金华 E-mail:yuziyi_yjh@hotmail.com
基金资助:
国家自然科学基金面上项目;国家自然科学基金面上项目

Effect of silencing PGC-1α on the directional differentiation of human stem cells from apical papilla

1. 南京医科大学
2. 无
3. Addiliated Hospital of Stomatology, Nanjing Medical University
4. Department of Endodontics, The Affiliated Stomatological Hospital of Nanjing Medical University, Jiangsu Province Key Laboratory of Oral Diseases, Jiangsu Province Engineering Research Center of Stomatological Translational Medicine
5.

Received:2023-02-09 Revised:2023-05-15 Online:2023-09-25 Published:2023-09-21

摘要/Abstract

摘要： 目的探讨过氧化物酶体增殖物激活受体γ辅激活因子1α（PGC-1α）在人根尖牙乳头干细胞（hSCAPs）定向分化中的作用。方法采用qRT-PCR检测hSCAPs矿化诱导0，3，7天后PGC-1α的表达水平。采用siRNA转染方法构建稳定低表达PGC-1α的hSCAPs，通过qRT-PCR和蛋白质印迹法检测PGC-1α的表达水平验证其敲低效率。采用碱性磷酸酶和茜素红染色，qRT-PCR观察低表达PGC-1α对hSCAPs定向分化的影响。结果PGC-1α的基因表达水平在hSCAPs定向分化过程中逐渐上调，且成功构建PGC-1α稳定低表达的hSCAPs。与对照组相比，低表达PGC-1α的hSCAPs其成牙/骨向分化相关指标表达水平下调，矿化结节减少，碱性磷酸酶表达量下降。结论沉默PGC-1α表达可抑制hSCAPs定向分化。

关键词: 受体γ辅激活因子1α, 根尖牙乳头干细胞, RNA干扰, 成牙/骨向分化

Abstract: ObjectiveTo explore the effects of silencing peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α)on the directional differentiation of human stem cells from apical papilla (hSCAPs). Methods Expression level of PGC-1α was detected by qRT-PCR after SCAPs were cultured in mineralized induction solution for 0, 3 and 7 days. HSCAPs with low expression of PGC-1α were constructed stably by siRNA transfection, and the knockdown efficiency of PGC-1α was verified by qRT-PCR and Western blot.Further, the effects of silencing PGC-1α on the directional differentiation of hSCAPs were observed by alkaline phosphatase and alizarin red staining and qRT-PCR. Results The expression level of PGC-1α was gradually up-regulated during the directional differentiation of hSCAPs, and hSCAPs with low expression of PGC-1α was successfully constructed. As compared with the control group,the expression level of indicators related with osteo/odontogenic differentiation in hSCAPs silencing PGC-1α were downregulated, mineralized nodules decreased, and the expression of alkaline phosphatase decreased. Conclusion Silencing PGC-1αcan inhibit the directional differentiation capacity of hSCAPs.

Key words: peroxisome-proliferator-activated receptor γ coactivator-1α , human stem cells from apical papilla, RNA interference, odonto/osteogenic differentiation

肖雅陈路遥顾婷杰方玉心李泽汉于金华. 沉默PGC-1α表达对人根尖牙乳头干细胞定向分化的影响[J]. 口腔生物医学, 2023, 14(3): 146-150.